Title of Invention

METHOD AND SYSTEM FOR DETERMINING AN EFFECTIVE AMOUNT OF LIGHT ENERGY TO DELIVER TO FLUIDS HAVING TARGETS FOR THE LIGHT ENERGY

Abstract A method of determining a fluid light energy value for delivery to a biological fluid comprising targets and non-target material, wherein an effective amount of light energy is desired to be delivered to said targets and wherein said non-target material attenuates said light energy destined for said targets, comprising the steps of: obtaining said target's effective light energy value; obtaining said fluid's average light energy factor; and calculating said fluid light energy value for delivery to said biological fluid, wherein determining the fluid light energy value is performed by one or more computer processors.
Full Text METHOD AND SYSTEM FOR DETERMINING AN
EFFECTIVE AMOUNT OF LIGHT ENERGY TO DELIVER
TO FLUIDS HAVING TARGETS FOR THE LIGHT ENERGY
FIELD OF THE INVENTION
This invention generally relates to determining an amount of light energy to deliver to
fluids, particularly partially transparent fluids, containing targets for the light energy, in order
to deliver an effective amount of light energy to the targets. The invention particularly relates
to phototherapy and photopheresis systems where an effective amount of light energy is
desired to be delivered to targets in biological fluids.
BACKGROUND OF THE INVENTION
Light irradiation or phototherapy has been widely used in the chemical and biological
sciences for many years. Ultraviolet (UV) light irradiation of blood was used in the 1930's,
40's, and 50's for the treatment of many conditions. These conditions included bacterial
diseases such as septicemias, pneumonias, peritonitis, wound infection, viral infections
including acute and chronic hepatitis, poliomyelitis, measles, mumps, and mononucleosis.
Phototherapy or light irradiation also includes the processes of exposing photoactivatable or
photosensitizable targets, such as cells, blood products, bodily fluids, chemical molecules,
tissues, viruses, and drug compounds, to light energy, which induces an alteration in or to the
targets. In recent years, the applications of phototherapy are increasing in the medical field.
These applications include the inactivation of viruses contaminating blood or blood products,
the preventive treatment of platelet-concentrate infusion-induced alloimmunization reactions,
and the treatment of both autoimmune and T-cell mediated diseases. Light irradiation
applications also include the irradiation sterilization of fluids that contain undesirable
microorganisms, such as bacteria or viruses.
Numerous human disease states, particularly those relating to biological fluids such as
blood, respond favbrably to treatment by visible or UV light irradiation. Light irradiation
may be effective to eliminate immunogenicity in cells, inactivate or kill selected cells,
inactivate viruses or bacteria, or activate desirable immune responses. For example;
phototherapy can be used as an antiviral treatment for certain blood components or whole
blood. {See PCT Application WO 97/36634 entitled Photopheresis Treatment of Chronic
HCV Infections). In this case, a pathogenic virus in a donated platelet concentrate can be
inactivated by UV light exposure.
Indeed, certain forms of light irradiation may be effective by themselves, without the
introduction of outside agents or compounds, while others may involvethe introduction of
specific agents or catalysts. Among the latter treatment techniques is the use of
photoactivatable drugs. In a particular application, it is well known that a number of human
disease states may be characterized by the overproduction of certain types of leukocytes,
including lymphocytes, in comparison to other population of cells which normally comprise
whole blood. Excessive abnormal lymphocyte populations result in numerous adverse effects
in patients including the functional impairment of bodily organs, leukocyte mediated
autoimmune diseases and leukemia related disorders many of which often ultimately result in
fatality.
Uses of photoactivatable drugs may involve treating the blood of a diseased patient
where specific blood cells have become pathogenic as a consequence of the disease state.
The methods generally may involve treating the pathogenic blood cells, such as lymphocytes,
with a photoactivatable drug, such as a psoralen, which is capable of forming photoadducts
with lymphocyte DNA when exposed to UV radiation.
A specific type of phototherapy is extracorporeal photopheresis (ECP). An
application of ECP is for the treatment of cutaneous T-cell lymphoma (CTCL). In an
example of this therapy, 8-methoxypsoralen (8-MOP), a naturally occurring light-sensitive
compound, is orally administrated to a patient prior to before ECP treatment. During the
ECP treatment, blood is withdrawn from the patient, anticoagulated, and the white cells are
separated by centrifugation and collected as a leukocyte enriched fraction, also known as the
buffy coat. The 8-MOP molecules in the blood enter the white blood cell nuclei and
intercalate in its double-stranded DNA helix.
In the extracorporeal circuit, UV light is directed at the leukocyte-enriched blood
fraction and promotes the photoactivation of the target 8-MOP molecules. The
photoactivated 8-MOPs alter the pathogenic leukocyte by cross-linking to the thymidine
bases and prevent the unwinding of DNA during transcription. The fluid containing the
altered leukocytes is then reinfused back into the patient. The reinfusion induces a
therapeutically significant delayed immune attack that targets antigens on the surface of both
irradiated and unirradiated leukocytes of the same pathogenic clones. See PCT Application.
WO 97/36581 entitled Photopheresis Treatment of Leukocytes, which is expressly hereby
incorporated herein by reference in its entirety. This PCT Application discloses the UVAR®
system for ECP. U.S. Patent Nos. 4,321,919,4,398,906, 4,428,744, and 4,464,166, each of
which is expressly hereby incorporated herein by reference in its entirety, also describe, inter
alia, methods for reducing the functioning lymphocyte population of a human subject using
photopheretic techniques.
ECP also has been shown to be an effective therapy in a number of autoimmune
diseases such as progressive systemic sclerosis (see A.H. Rook et al., ARCH. Dermatol.
128:337-346 (1992)), inflammatory bowel disease, rheumatoid arthritis (see S. Malawista, et
al., Arthritis Rheum. 34:646-654 (1991)), and juvenile onset diabetes mellitus (see J.
Ludvigsson, Diabetes Metab. Rev. 9(4):329-336 (1993)), as well as other T-cell mediated
phenomena including graft-versus-host disease (see Rosseti et al., TRANSPLANT 59(1): 149-
151 (1995)), and organ allograft rejection after transplantation (see A.H. Rook, et al., J. CLIN.
Apheresis 9(l):28-30 (1994)). The ECP treatment preferably results in a highly specific
immune response against aberrant T-cells as well as removal of pathogenic antibodies and
circulating immune complexes.
A difficulty inherent in light irradiation or phototherapy techniques when used in the
irradiation of fluids and/or their target components, however, is that often times the fluid is
not completely transparent to light, e.g., the fluid itself is not entirely transparent and/or the
fluid contains material (e.g., non-target material) that is not entirely transparent to light.
Material that is not completely transparent to light energy attenuates the irradiance of the
light. This phenomenon is particularly undesirable in phototherapy or photopheresis
applications since some targets in the fluid will receive light that is attenuated by the non-
transparent material. This attenuation makes it difficult to predict how much light energy
should be delivered to the fluid to provide a desired amount of light energy to targets in the
fluid.
Another source of light attenuation in fluids is stacking. Stacking occurs in a fluid
when material or targets in the fluid are not distributed uniformly on the fluid surface but
rather are located at different depths throughout the fluid. Therefore, for instance, targets in
the outer most layer of the fluid, closest to the irradiating light source, may be exposed to
incident light intensity, while the targets below the surface layer may receive attenuated light
energy.
Furthermore, the shapes of non-transparent material in the fluid and their alignment
can be a cause of light attenuation. For example, in photopheresis applications, non-targets in
the biological fluid may include red blood cells, which have discoid shapes with depressions
at the middle. When red blood cells are aligned parallel to the light energy source during
irradiation, their attenuation of light is minimized. However, when red blood cells are
aligned perpendicular to the light energy source during irradiation, their attenuation of light is
maximized. Since the alignment of such fluid material is usually not predictable, it is
presently difficult to accurately determine how much light energy should b& delivered to the
biological fluid in order to deliver a desired amount of light energy to each target in the fluid
and overcome the light attenuation caused by the alignment of the material.
The CTCL EC? methodology referenced in PCT Application WO 97/36581 can be
used to illustrate these exemplary light attenuation characteristics. The buffy coat suspension
usually contains some red blood cells and platelets due to inefficiencies inherent in the cell
separation techniques utilized. Since the buffy coat suspension, red blood cells and platelets
are not completely transparent, they can attenuate the light energy during irradiation. Also,
since the fluid's thickness during irradiation can support target white blood cells at different
depths, stacking is present. Finally, the alignment of red blood cells in the fluid containing
the buffy coat may attenuate the light energy.
With CTCL ECP, the desired amount of light energy for delivery to targets may be
result-based, e.g., delivering enough light energy to the target white blood cells to produce a
gradual death rate culminating in at least fifty (50) percent of treated, irradiated white blood
cells dead after day six (6) of irradiation. Yet, the fluid's non-transparent qualities presently
make it difficult to accurately calculate the amount of light energy required to deliver to the
fluid, in order to achieve the desired result.
A conventional way to reduce the effect of the attenuation of light in such applications
is to constantly agitate the fluid during irradiation. Agitation assists to produce uniform
exposure of the targets to the light energy, yet it does not directly address all the light
attenuating factors present in such applications. See PCT Application WO 98/22164, entitled
Blood Product Irradiation Device Incorporating Agitation, which is expressly incorporated
herein by reference.
It is therefore desirable to have a system for determining an effective amount of light
energy to deliver to fluids containing targets for the light energy, in order to deliver an
effective amount of light energy to the targets and, more particularly, to have a system
applicable to phototherapy and ptiotoplheresis systems for determining an effective amount of
light energy to deliver to a biological fluid containing targets for the light energy where an
effective amount of light energy is desired to be delivered to the targets.
The present invention relates to methods and systems for determining the effective
amount of light energy for delivery to a fluid containing targets, and delivering said light
energy to the targets. In a specific embodiment, the fluid is a biological fluid. Specifically,
the fluid light energy value (FLEV) may be calculated by obtaining the target's effective light
energy value (TELEV) and the average light energy factor of the fluid (ALE Factor). In a
specific embodiment, a computer processor may be used to determine the FLEV.
In a specific embodiment, the fluid containing the targets is a biological fluid. More
preferably, the biological fluid comprises leukocyte-rich buffy coat. The leukocyte-rich
buffy coat may be treated with a light energy activatable drug. More preferably, the buffy
coat may be treated with 8-MOP. In another embodiment of the present invention, the fluid
is a homogenous biological fluid. The biological fluid may also comprise non-target
materials. These non-target materials may attenuate the light energy, and affect calculation
of the FLEV- Non-target materials may consist of red blood cells. Further, the light energy
delivered to the targets may be UV light energy. More preferably, the light energy is
ultraviolet A (UVA) light energy.
In a specific embodiment, the effective light energy value of the targets may be
obtained by accessing an effective light energy value table. In another embodiment, the
effective light energy value of the targets may be obtained by placing the targets in fluid and
irradiating the fluid with sample light energy values. The selected fluid may limit the
attenuation of the delivered light energy. In a specific embodiment, the fluid may consist of
saline. More specifically, leukocyte-rich buffy coat targets may be placed in saline and
irradiated to identify a light energy value whereby a desired percentage of the leukocytes will
gradually die over the course of a specified time after exposure to the light energy. In yet
another embodiment, the selected fluid may consist of plasma. Sample biological fluids may
be obtained from donors. The targets in the sample fluids may then be irradiated with sample
light energy values to identify the effective light energy value. In a specific embodiment, a
computer processor may be used to determine the effective light energy value of the targets.
The fluid's average light energy factor may be determined by accessing a light energy
factor table. In a specific embodiment, a computer processor may be used to determine the

average light energy factor.
In another embodiment of the present invention, the average light energy factor may
be calculated from the measurements of an average light energy value at a unit surface area of
the targets in the biological fluid and a light energy value at an incident surface of the
biological fluid film. In a specific embodiment, the average light energy at unit surface area
of the targets in the biological fluid may be obtained by accessing an average light energy at
unit surface area table. The light energy value at an incident surface may also be obtained by
accessing a light energy value at an incident surface table. These values may also be directly
calculated.
In a further embodiment, the average light energy factor may be calculated from the
measurements of a thickness ratio and a light transmittance value of a known fluid thickness.
The thickness ratio may be obtained by accessing a thickness ratio table. The irradiation
period may be obtained by accessing a light transmittance value of a known fluid thickness.
In another embodiment, the thickness ratio may be calculated from the uniform thickness for
said biological fluid and the thickness for said non-targets. Further, the uniform thickness for
the biological fluid may be obtained by accessing a uniform thickness table, while the
thickness for non-targets may be obtained by accessing a non-target thickness table.
In another embodiment of the present invention, the average light energy factor may
be calculated from the measurements of a thickness ratio and the red blood cell percentage of
the biological fluid. The red blood cell percentage may be obtained by accessing a red blood
cell percentage table.
Another method for calculating the average light energy factor may utilize the
measurements of the uniform thickness of the biological fluid and the red blood cell
percentage of the biological fluid. The equations used in this method may preferably be used
for red blood cell concentrations in the biological fluid of up to about twenty (20) percent,
and more preferably for red blood cell concentrations of up to about seven (7) to eight (8)
percent.
In one embodiment, theoretical stacking of red blood cells may not occur. In another
embodiment, stacking of red blood cells may occur and a factor may be obtained. This factor
may, in a particular embodiment, be between 1 and 2, and more particularly about 1.5.
In a further embodiment, the irradiation time period required by a light energy source
to deliver the FLEV may be calculated once the target's effective light energy value and the
fluid's average light energy factor have been determined using one of the methods of the
present invention and used to calculate the FLEV. The irradiation time period may be
calculated from measurements of a volume of biological fluid value, a percent of red blood
cells value and a decay life value.
In another embodiment of the present invention, a computer system may be used to
determine the FLEV. This computer system may comprise a processor, memory and a
computer process. More specifically, the computer process may comprise an obtainer
configured to obtain the effective light energy value of the target, an obtainer configured to
obtain the average light energy factor of the fluid and/or a calculator configured to calculate
the FLEV. In a specific embodiment, the calculator used to calculate the FLEV may also be
configured to calculate an irradiation time period over which the FLEV is delivered to the
targets. The calculator used to calculate the FLEV may also contain an obtainer to obtain a
decay life value for the light energy source. The calculator may also contain an obtainer to
obtain a volume of biological fluid value and an obtainer to obtain a percent of red blood
cells value.
In a specific embodiment, the obtainer configured to obtain the effective light energy
value of the targets may include an accessor configured to access a light energy factor table.
In another embodiment, the obtainer configured to obtain the effective light energy value of
the targets may include an obtainer configured to obtain the average light energy value at a
unit surface area of the targets, an obtainer configured to obtain a light energy value at an
incident surface of the biological fluid and/or a calculator configured to calculate the average
light energy factor. More preferably, the obtainer configured to obtain a light energy value at
an incident surface of the biological fluid may contain an accessor configured to access an
average light energy value at an incident surface of the biological fluid table, and/or an
accessor configured to access an average light energy value at unit surface area table.
The obtainer configured to obtain an average light energy factor may contain an
obtainer configured to obtain a thickness ratio, an obtainer configured to obtain a light
transmittance value of a known fluid thickness and/or a calculator configured to calculate the
average light energy factor for the biological fluid. More preferably, the obtainer configured
to obtain a thickness ratio may contain an accessor configured to access a thickness ratio
table, and the obtainer configured to obtain a light transmittance value of a known fluid
thickness may contain an accessor configured to access a light transmittance value of a
known fluid thickness table.
In a further embodiment, the obtainer configured to obtain the thickness ratio may
include an obtainer configured to obtain a uniform thickness for the biological fluid, an
obtainer configured to obtain a thickness for the non-targets and/or a calculator configured to
calculate the thickness ratio. More preferably, the obtainer configured to obtain a uniform
thickness for the biological fluid may contain an accessor configured to access a uniform
thickness table, and the obtainer configured to obtain a thickness for the non-targets may
contain an accessor configured to access a non-target thickness table.
In a further embodiment, the obtainer configured to obtain the average light energy
factor may include an obtainer configured to obtain a red blood cell percentage for the
biological fluid. More preferably, the obtainer configured to obtain a red blood cell
percentage may contain an accessor configured to access a red blood cell percentage table.
In another embodiment of the present invention, the obtainer configured to obtain the
thickness ratio may include an obtainer configured to obtain a uniform thickness for the
biological fluid, an obtainer configured to obtain a thickness for the non-targets and a
calculator configured to calculate the thickness ratio. More preferably, the obtainer
configured to obtain the uniform thickness may contain an accessor configured to access a
uniform thickness table, and the obtainer configured to obtain the thickness of the non-targets
may contain an accessor configured to access a non-target thickness table.
In a further embodiment, the obtainer configured to obtain the average light energy
factor may include an obtainer configured to obtain a red blood cell percentage for the
biological fluid. The computer system may further include an obtainer configured to obtain
the red blood cell stacking factor. In a particular embodiment, the stacking factor may be
between 1 and 2. More particularly, the stacking factor, may be 1.5.
The present invention also relates to a computer readable medium containing
instructions for controlling a computer system used to perform the methods described herein
for determining a fluid light energy value for delivery to a biological fluid comprising targets,
wherein an effective amount of light energy is delivered to the targets.
Methods and articles of manufacture consistent with the present invention may
involve the functions and operations performed by the described systems and the components
thereof.
Other objectives, features, and advantages of the present invention will become
apparent from the following detailed-description. The detailed description and the specific
examples, while indicating specific embodiments of the invention, are provided by way of
illustration only. Accordingly, the present invention also includes those various changes and
modifications within the spirit and scope of the invention that may become apparent to those
skilled in the art from this detailed description.
BRIEF DESCRIPTION OF THE FIGURES
The accompanying figures, which are incorporated in and constitute a part of the
specification, illustrate embodiments of the invention and, together with the description, serve
to explain the objects, advantages and principles of the invention.
Figure 1 is a diagram 100 of a photopheresis system according to an implementation
of the present invention.
Figures 2 A and 2B are a flow diagram 200 of the steps performed by a photopheresis
system according to an implementation of the present invention.
Figure 3 is a diagram 300 of a computer system for controlling the photoactivation
device according to an implementation of the present invention.
Figure 4 is a flow diagram 400 of the steps performed by the photoactivation program
314 when requested to deliver light energy to fluid according to an implementation of the
present invention.
Figure 5 is a flow diagram 500 of the steps performed by the photoactivation program
314 when calculating the target's effective light energy value according to an implementation
of the present invention.
Figure 6 is a flow diagram 600 of the steps performed by the photoactivation program
314 when calculating the average light energy factor for the fluid according to an
implementation of the present invention.
Figure 7 is a flow diagram 700 of the steps performed by the photoactivation program
314 when using an analytical equation to calculate the average light energy factor for the
fluid according to an implementation of the present invention.
Figure 8 is a flow diagram 800 of the steps performed by the photoactivation program
314 when using calculate the average light energy factor for fluids containing red blood cells
as non-targets according to an implementation of the present invention.
Figure 9 is a flow diagram 900 of the steps performed by the photoactivation program
314 when using a stacking equation to calculate the average light energy factor for the fluid
according to an implementation of the present invention.
Figure 10 is a graph of the average light energy factors as a function of percent
hematocrit at different fluid thicknesses according to an implementation of the present
invention.
Figure 11 is a table providing an exemplary single lamp decay value over time.
Figure 12 is a graph of average single lamp irradiance measured at a distance of 25
cm from the center line of a lamp over time.
The following definitions are not meant to be limiting in nature and serve to provide a
er understanding of certain aspects of the present invention,
nitions:
Target — Targets include photosensitive or photoactivatable materials that undergo a
ge when exposed to light energy. Accordingly, targets maybe manipulated, altered,
alated and/or activated when exposed to light energy. Targets include, but are not
limited to, biological targets such as red blood cells, white blood cells, platelets, protein.
factors, viruses, bacteria, parasites, DNA, RNA, toxins, and drug compounds. Targets
exposed to light energy may also interact with other materials or targets.
Phototherapy — Phototherapy includes procedures where photosensitive,
photochangeable or photoactivatable targets are exposed to light energy.
Fluid — Fluids include substances that may be used as carriers of targets. Examples
of fluid include spinal fluid, cells, and other fluids compatible with a target such as phosphate
buffered saline, plasma, etc., and combinations thereof. The fluid may include non-targets
and may be biological in nature.
Non-target — Non-targets include material that attenuates light energy, yet are not
the intended targets for light energy. Non-targets include red blood cells and platelets.
Biological Fluid — Biological fluids include fluids that carry targets and/or non-
targets and that have the capacity to support biological targets. Biological fluids may include
whole blood, plasma, synovial fluid, amniotic fluid, and spinal fluid, in addition to carriers
such as saline or other known media, preferably compatible with biological organisms such
as cells and tissues, and combinations thereof.
Photopheresis -- A type of phototherapy in which fluid is extracted from a donor,
exposed to light energy, and returned to the donor. In a particular embodiment, the extracted
fluid, such as whole blood or portions of whole blood (such as buffy coat), may contain
targets. CTCL ECP is an example of photopheresis.
Photoactivation - Photoactivation is a process in which a target is changed (e.g.,
manipulated, altered, stimulated, or activated) by exposure to light energy. An example of a
target undergoing photoactivation is the drug 8-MOP used in CTCL ECP which, previous to
photoactivation, is inert. Exposing this drug compound to light energy activates it to a form
that can cross-link lymphocyte DNA.
Light Energy — Light energy is the form of energy that reacts with targets, such as
biological or chemical targets. An example of light energy used in phototherapy applications
is UV light and, more specifically, UVA light in CTCL ECP methodology.
Desired Result - A desired result is an outcome for light energy manipulated targets.
In the CTCL ECP context, for example, a desired result might be to have a specific
percentage of irradiated leukocytes gradually die over a specific time period after exposure to
the light energy.
TELEV — The Targets' Effective Light Energy Value is the light energy value
delivered to the targets, preferably calculated in a medium or fluid that contains essentially no
other light attenuating material, that produces a desired result.
ALE Factor — The Average Light Energy Factor compares the amount of light
energy present at the incident surface of the fluid with the amount of light energy at the
surface of the targets within the fluid.
FLEV - The Fluid's Light Energy Value is the amount of light energy delivered to
the fluid to maximize the probability that targets receive their TELEV.
Uniform Fluid Thickness — Uniform fluid thickness is the fluid thickness where the
light irradiation of targets occurs.
Non-Target Thickness — The non-target thickness is the thickness of the non-target
material that is the dominant light attenuating non-target material in a fluid.
Thickness Ratio — The thickness ratio is the ratio of the uniform thickness of the fluid
to the average thickness of non-targets in the fluid.
Irradiation Period — Irradiation period is the time period that the light energy source
irradiates the fluid containing the targets.
Reference will now be made in detail to implementations of the present invention as
illustrated in the accompanying drawings. Wherever possible, the same reference numbere
will be used throughout the drawings and the following description to refer to the same or
like parts.
Light irradiation methodologies, as discussed above, involve the delivery of light
energy to a target to achieve a desired result. The targets may be carried in a medium (e.g., a
fluid) during light irradiation. In a particular context of the present invention, the amount of
light energy delivered to targets in a fluid that contains essentially no non-target light
attenuating material, in order to achieve the desired result, is the TELEV. Indeed, non-target
materials may also be present in the fluid, which may result in the attenuation of the light
energy that is desired to be delivered to the targets. Accordingly, the present invention, inter
alia, accounts for the light attenuation of the non-target material present in the fluid by
determining the FLEV so that the TELEV may be delivered to the target material.
In a specific application of the present invention, phototherapy systems involve
irradiating targets, such as cells or a drug within a cell, with light energy. When the targets
are microscopic or unable to stand-alone, a carrier fluid may be used to deliver the targets for
irradiation.
The amount of light energy required by a target may be based on the result desired.
For example, in CTCL ECP it may be desirable to have a certain percentage of the white
blood cells die gradually over a specific period of time after light irradiation treatment (e.g.,
at least fifty (50) percent of the white blood cells gradually die within six (6) days after
irradiation). See PCT Application WO 97/36581. This light energy value required to
produce a desired result (e.g., a desired percentage of the targets gradually die over a
specified time after exposure to light energy) is the TELEV. There are a number of
conventional approaches that can be used to obtain the TELEV. Some of these approaches
are discussed later in more detail. Indeed, TELEV values may be predetermined and may be
available in the memory of a computer system used with the present invention, e.g., in a look-
up table.
Since the material in a fluid may attenuate light energy otherwise desired to be
delivered to targets in the fluid, the fluid requires additional light energy exposure to
maximize the probability that targets in the fluid receive their TELEV. The amount of light
energy needed for delivery to the fluid to maximize the probability that the targets receive the
TELEV is referred to as the Fluid's Light Energy Value (FLEV). The FLEV is based, in
part, on the light attenuation characteristics of the fluid and material therein, and may be
determined by the methods and systems of the present invention.
As discussed above, light attenuation in the fluid may occur for numerous reasons.
For example, attenuation may occur if the fluid being irradiated contains target and/or non-
target material that are not entirely transparent. Also, attenuation may occur ifthe fluid
sample being irradiated supports layers of targets and/or non-targets. In addition, the shape
and alignment of the individual targets and/or non-targets may influence the quantity of light
attenuation.
In one embodiment of the present invention, the FLEV can be calculated by
determining the TELEV and the percent of incident light energy that will be delivered to an
average unit area of fluid. This percent is referred to as the fluid's Average Light Energy
Factor (ALE Factor). Once the ALE Factor is known, the FLEV can be determined as
follows:
(1.0)
For example, it may be determined that one (1) Joule of UV light energy delivered to
targets will produce a desired result (TELEV). However, due to attenuation of the light
energy in the fluid (e.g., via the presence of non-transparent material in the medium
containing the targets or stacking), the light energy reaching the targets in the fluid is reduced
and, thus, approximately 0.1 Joule of UV light energy actually reaches the targets. Thus, the
ALE Factor is 0.1, i.e., only ten (10) percent of the light energy delivered to the surface of the
fluid will actually reach (on average) all the targets. Thus, applying equation 1.0, 10 Joules
(FLEV) of light energy is required to be delivered to the surface of the fluid to ensure that the
targets (on average) receive 1 Joule of light energy (the desired result).
In a particular embodiment of the invention, the ALE Factor may be determined by
dividing the light energy delivered at the unit surface area of the targets, Ea (Joules/cm2), by
the incident light energy delivered at the incident surface of the fluid, Eo (Joules/cm2):
(1.1)
The following provides exemplary means for determining the ALE Factor, taking into
consideration the light attenuating characteristics of the fluid and its components. By way of
example, in CTCL ECP applications when a buffy coat suspension with a uniform film
thickness (D) is irradiated by an UVA light with an incident irradiance (Io) at the fluid film
surface (mW/cm2 ), the Eo delivered at the fluid surface during a given irradiation period (t) is
expressed by equation 1.2:
(1.2)
Buffy coat suspension is partially transparent to the UVA light. Accordingly, this
fluid attenuates the irradiance of the light at a given point inside the fluid. The degree of
attenuation is a function of absorptivity of the fluid and the light penetration depth from the
fluid surface.
Assuming Beer's law, the light transmittance (T1) of the fluid between its incident
surface and a given point in the fluid at a distance (DO can be expressed as:

where a is the light absorptivity of the fluid (cm2/gr) and c is the concentration of UVA
absorbing component in the fluid (gr/cm3).
Equation 1.3 can be expressed as:

Where Dn is the distance from the incident surface of the fluid and n is Dn/D1:

In addition, the irradiance (In) at the distance of Dn from the incident surface is:

The average irradiance value (la) over the whole range of the fluid film thickness (Dr)
can be calculated by integrating over the range of the fluid depth and dividing it by the film
thickness:

where N = D1/D1 and the ratio of la to Io is:

Integrating over the film thickness the ratio becomes:
(1.9)
One thus arrives at the following analytical equation:
(2.0)
where N is the ratio of the uniform film thickness D (cm) to the non-target non-transparent
material thickness D1 (cm), and T1 is the light transmittance of the light through the fluid,
when the fluid has a fluid thickness equal to the thickness of the dominant non-target. A non-
target is dominant when compared to any other non-targets. It is the predominant light
attenuator. The accuracy of this calculation may be maximized in situations where the target
and dominant non-target material in the fluid are uniformly distributed throughout the fluid,
e.g., by stirring.
Equation 2.0 is particularly applicable to partially transparent fluids and, in particular,
can be used in photopheresis applications to estimate the average amount of UVA light
energy delivered to white blood cells in a well stirred buffy coat suspension. In a specific
embodiment, when the application is used with fluids containing red blood cells (with a
thickness of about 2*10-4 cm) as dominant non-target, light attenuating material, equation
2.0 becomes:
(2.1)
where H is the hematocrit value of the fluid.
An additional exemplary way to determine the ALE Factor, preferably when the fluid
comprises a dominant attenuating non-target such as red blood cells, is to use the following
stacking equation:
(2.2)
where C is the percent of non-targets in the fluid and D (cm) is the fluid thickness. Y is a
dimensionless number that represents the geometric shape of the non-target and the stacking
factor. The stacking factor is also a dimensionless number that represents the theoretical
amount of physical stacking that takes place within the fluid by the non-targets. In ECP
applications, for example, the stacking factor may be a number between 1 and 2. Means for
obtaining a stacking factor are described in detail supra. When the non-target is
geometrically spheroid, the equation for Y is:
(2.3)
where R (cm) is the average radius of the non-target, d (cm) is the average thickness of the
non-target, and S is the stacking factor.
When red blood cells are the dominant attenuating non-target in a buffy coat
suspension, equation 2.2 becomes:
(2.4)
where H is the hematocrit value for 1 ml of buffy coat suspension.
The following provides an example of how the stacking equation and stacking factor
may be derived. Turning to the exemplary CTCL ECP methodology, red blood cells have a
diameter of about 8 * 10-4 cm and thickness of about 2 * lO-4cm. There are two extreme
cases of orderly aligned situations for the red blood cell distribution in the buffy coat
suspension. The first is where all RBC's are evenly distributed in the cube and aligned in
such a way that their interference to the UVA irradiation is maximized. In another words, the
discoid sides of all RBC's are in perpendicular position against the incoming UVA light rays.
The second is where all RBC's are evenly distributed in the cube and aligned in such way
that their interference to UVA irradiance is minimized. In another words, the discoid sides of
all RBC's are in parallel position against the incoming UVA light rays.
In the CTCL ECP context, RBC's are preferably randomly distributed in the
suspension and the effect of the interference could be somewhere between these two
theoretical situations. Here, a one cubic centimeter (or unit volume) of well-mixed buffy coat
suspension with UVA light irradiated on one side only is considered. In addition, in these
two cases it was assumed that no RBC's were stacked against each other, i.e. no rouleaux
formation, because of low hematocrit in buffy coat suspensions.
Considering a situation where light interference by RBC's is maximized, each cubic
centimeter (ml) of the buffy coat suspension could be sliced into 1/d slices where d is the
thickness of the red blood cells. Accordingly, the number of RBC's in each slice is:
(2.5)
where C is the RBC concentration (number of cells/ml) in the buffy coat suspension. Thus,
the maximum possible fractional area (Fa) that could block UVA irradiation in a given slice
is:
(2.6)
where R is the radius of the RBC.
The theoretically minimum number of slices that is required to block the UVA light
completely in one square centimeter of irradiated surface area thus is 1/Fa. In order to achieve
this, no red blood cell should be shielded behind another red blood cell. The total number of
the slices in the cube is 1/d. Therefore, in one cubic centimeter volume of the buffy coat
suspension, there are (l/d)/(l/Fa) times of (1/Fa) slices. It follows that one cubic centimeter
(or unit volume) of buffy coat suspension contains a total number of slices that can
theoretically shield (l/d)/(l/Fa) times of one square centimeter area (or unit area) from the-
UVA light. Substituting for Fa in equation 2.6:
(2.7)
In this instance, no red blood cells are shielding other red blood cells from UVA light.
For example, if the hematocrit is 5%,' the first slice will block 5% of the UVA irradiation and
the second slice will block additional 5%, and so on. The last layer in the 1/Fa slices will
block the last remaining 5% of the UVA light, thereby blocking the light completely. Under
this condition about slightly less than half of the fluid, including the target cells within, is
irradiated by the UVA light; the remaining portion of the fluid is shielded from the light by
the red blood cells.
Another situation is where all red blood cells in a slice are located behind other red
blood cells in the slice in front of it. For instance, if the hematocrit is 5%, only 95% of the
first slice will pass the light. Since all red blood cells in the second and slices behind it are all
located behind the red blood cells in the first layer, there is no further blocking of the light
and 95% of all fluid in (1/Fa) slices, almost twice as much as the former case, will receive the
UVA irradiation. Therefore, incorporating a simple stacking factor (S), equation 2.7 can be
rewritten as:
(2.8)
The value of the stacking factor, S, in ECP applications, thus may be between one and two.
Following a similar analysis, equation 2.8 becomes:
(2.9)
where d' = 2*R.
Equations 2.8 and 2.9 represent two opposite extreme cases of RBC light attenuation.
In practice, RBC attenuation of light is somewhere between these two extremes. If we take
the average of these extreme cases as an estimate for the situation in practice, the equation
becomes:

For human blood buffy coat suspensions we can approximate R = 4 * 10-4 cm and d =
2*10 cm for red blood cells. Accordingly, equation 3.0 becomes:

Equation 3.1 represents multiples of number of slices, which can block completely the
incoming UVA light through one square centimeter area, in one cubic centimeter volume.
Assuming the buffy coat suspension inside this one cubic centimeter volume (or unit volume
is well mixed, the UVA energy delivered to the target cells through the one square centimetei
(or unit area) window may be expressed as:

where Ea = UVA energy delivered per unit area, Joules/cm
Ev = Eo*A/V, UVA energy delivered per unit volume, Joules/ml
Eo = Io*t, Incident UVA energy delivered per unit area, Joules/cm
Io = Incident irradiance, Joules/cm2-sec.
t = Irradiation time, seconds.
V = A*D, Irradiated volume, ml.
A = Irradiation area, cm2
D = Buffy coat film thickness, cm
C = Red blood cell concentration, -1.1 * H * 108 cells/ml
H = Hematocrit of the buffy coat suspension, %
S = Stacking factor, dimensionless number between 1 and 2.
Substituting S = 1.5, the average of 1 and 2, as an estimate and C = 1.1*H*108,
equation 3.2 becomes:
(3.3)
(3.4)
Equations 2.0 and 2.4, when applied to a fluid containing red blood cells as the
dominant attenuating material, predict almost identical ALE factors up to a red blood cell
concentration of about 20%, as represented in Figure 10. At higher red blood cell
concentrations, where the theoretical condition assumed in the stacking equation deviates
further from the real situation, the difference between the two equations becomes predictably
greater. Indeed, at red blood cell concentrations of over 20% it may be more appropriate to
use equation 2.0. At extremely low red blood cell concentrations (e.g., less than 0.2%),
where the attenuation caused by the plasma component of the suspension itself is no longer
negligible in comparison with the attenuation produced by the red blood cells, equation 3.4
may lose some of its accuracy.
Another method for calculating the ALE Factor may utilize the measurements of the
uniform thickness of the biological fluid and the red blood cell percentage of the biological
fluid. The equations used for this method can be preferably utilized with red blood cell
concentrations in the buffy coat suspension of up to twenty (20) percent, and most preferably
used with a red blood cell concentration of up to between seven (7) and eight (8) percent
Once the FLEV is calculated, an additional calculation based on the specific light
delivery system may be made. The delivery system calculation determines what irradiation
time period is needed to deliver the FLEV to the fluid, taking into consideration a variety of
factors related to the light source and its present ability to deliver light. This calculation may
preferably take into consideration factors such as the shape of the light source, the lamp
decay over time, the size of the light beam, and the volume of the fluid being irradiated.
The variable L (mW/cm2) accounts for decay of the output of the light source over
time and depends upon the properties of the lamp source used, preferably measured at a fixed
position from the lamp center line. By way of example, L may be determined by taking
hourly measurements of an exemplary lamp over the lamp's lifetime. As time progresses, the
lamp intensity decreases. In a specific embodiment, once the hourly measurements are
plotted, an equation can be created to match the measurements. Then, the equation can be
utilized to determine the value of L by merely knowing how many lamp hours have been
used. In an alternate embodiment, a database containing the lamp life measurements can be
directly accessed.
For example, in a particular embodiment of the present invention, Figure 11
represents, in a prototype look-up table, the L value (mW/cm2) over 150 hourly
measurements for a lamp utilized in the UVAR® system taken 25 cm from its center. These
measurements result in the following single lamp irradiance decay equation:
(3.5)
The L value allows one to adjust for lamp life in determining the length of time the
light source irradiates the targets to achieve the desired result. Based on the L values of
Figure 11, an exemplary single lamp irradiance decay equation is determined where a equals
0.78552878, b equals -0.00059106023, and c equals -0.032384473. This equation, as well as
the table for L values for the light source utilized, may be stored and accessible for example,
in system memory or in a look-up table.
In the exemplary UVAR® system, the photoactivation chamber is located between
two banks of UVA lamps and the buffy coat suspension is recirculated through a serpentine
path inside the photoactivation chamber. The blood film thickness in the chamber is the
same, about 1.4 mm thick. At this blood film thickness, with hematocrit value at least around
few percent, the irradiating UVA light is completely absorbed by the blood film and the total
amount of UVA energy delivered to the each ml of circulating buffy coat suspension can be
calculated. This value is 255 Joules/ml in the UVAR® system.
The irradiance of UVA light reaching the surface of the target cells in the suspension
is attenuated by the red blood cells in the light path. The red blood cell is almost completely
opaque to the UVA light. Under these conditions, it is reasonable to assume that the
attenuation of the irradiance is inversely proportional to the red blood cell concentration in
the light path. The concentration of white blood cells is about one order of magnitude less
than that of red blood cells and also the white blood cell is much less opaque to UVA light
than red blood cells. Therefore, the amount of attenuation caused by the white blood cells
will be insignificant and may be ignored in the derivation of irradiation time equation.
The total amount of UVA energy delivered to the each ml of the circulating buffy coat
suspension can be expressed as:
Ev = k*H (3.6)
where Ev = Total amount of UVA energy delivered per unit volume, Joules/ml
k = Proportional constant
H = Hematocrit.
In the UVAR® system, the value of Ev is 255 Joules/ml and the average hematocrit
value is about 3.5%. Therefore, k = 255/3.5.
UVA energy is delivered through the irradiation chamber and to the surface of the
buffy coat suspension film inside the irradiation chamber while the buffy coat film is flowing
inside the irradiation chamber. The total amount of UVA energy delivered to the total
volume of the buffy coat suspension can be calculated by multiplying the irradiance at blood
film surface (through the chamber wall), the irradiation period and the irradiated blood film
area. Also, the UVA energy delivered to a unit volume, Ev, can be expressed by dividing the
total amount of UVA energy delivered divided by the total buffy coat suspension volume.

where Ev = UVA energy delivered per unit volume, J/ml
Io = UVA irradiance at blood film surface, mW/cm2
A = Area of blood film irradiated inside irradiation chamber, 1330 cm2
t = Irradiation period, minutes
V = Total buffy coat suspension volume in the circulation loop, ml.
The multiplication factors, 1000 and 60, may be utilized for unit correction from milliwatts to
watts and from minutes to seconds.
Combining equations 3.6 and 3.7, and substituting k = 255/3.5 and A = 1330 cm , the
irradiation period can be expressed as:

The equation for the average irradiance value, Io, of the UVA light at blood film
surface inside the irradiation chamber can be derived as follows.
The UVA light reaching the surface of the blood film inside the UVAR® irradiation
chamber comes from a light set consisting of nine (9) lamps. In the instrument light box, the
UVA light passes through UVA transparent glass and the acrylic irradiation chamber wall
before it reaches the blood film. Also, the UVA output is not uniform along the length of the
tubular fluorescence UVA lamp. The output is higher in the middle section of the lamp and
lower near the ends of the lamp. Therefore, the average irradiance value of the UVA light
reaching the blood film can be obtained by measuring the irradiance at points along the light
set and calculating their average value. However, since lamp output decays over time, it is
extremely difficult to measure all points simultaneously at a given lamp time. As described
below, this problem was resolved by the relationship of this average value to the average
single lamp irradiance value at one fixed point that can be measured quickly.
Figure 12 shows the average UVA irradiance value of six (6) single lamps measured
at mid-point and at a 25 cm distance from the lamp center line as a function of lamp life. The
irradiance value decays very rapidly at the beginning and decreases more gradually as the
lamp life increases. After around 60 hours of use, the lamp output decays rather slowly and it
allows enough time to measure points in the light set and calculate the average irradiance
value. The irradiance measurements were made at the 61.5 hour point and the 150 hour point
in several light sets. The values were 15.11 and 11.19 mW/cm2 at 61.5 hours and 150 hours,
respectively. The ratios of these average irradiance values in the light box and the average
single lamp irradiances at corresponding lamp life were calculated. The ratios were 23.9 at
the 61.5 hour point and 21.9 at the 150 hour point, resulting in the average value of 22.9.
Io in Equation 3.8 can be expressed as:
(3.9)
where k = Irradiance ratio of the light box and single lamp, 22.9
L = Single lamp irradiance, mW/cm2
T = Percent UVA transmittance of acrylic irradiation chamber, 92%.
Substituting equation 3.9 for Io in equation 3.8 and actual values for corresponding
variables, the irradiation time equation 3.8 becomes:
(4.0) -
where L is the single lamp irradiance expressed as a regression line equation based on
measured data points shown in Figures 11 and 12.
In an exemplary UVAR® system used in CTCL ECP applications, the following
equation 4.1 is used via the methods and systems of the present invention to determine
irradiation times:
(4.1)
where tmin = Irradiation time, minutes
V = Volume of the fluid in the treatment/recirculation loop, ml
H = Hematocrit
T = 92 (% transmittance of irradiation chamber)
k = 23.9 (a constant based on a ratio of the intensity of one lamp measured at one
point in the fluid to the intensity of the entire lamp set in the UVAR®
system).
Correcting for time in seconds, gives:
(4.2)
Inserting constants gives:
(4.3)
Collecting constants gives:
(4.4)
Referring to Figure 11, and using the following parameters:

The L value at lamp life of 2 hours is 7625 in Figure 11. The L value at lamp life of 3
hours is 7488. Linear interpolation using integer arithmetic gives:

The UVAR® instrument, in a specific embodiment, uses two lamp banks. The lamp
ages of these banks can differ, and theoretically, so can their irradiation time tables. To
account for this, the complete calculation is preferably run twice, once for each lamp bank,
and the values may be averaged. This value is the photoactivation time. Once the calculation
is run the time remaining is preferably immediately decremented by the amount of time the
UV lamps have already been on in the UVAR® system.
Once the irradiation time period is calculated, the present invention contemplates the
additional step of delivering the light energy, for that period of time, to the fluid containing
targets. In a particular embodiment of the present invention, the system then may instruct the
photoactivation device to deliver the FLEV to the fluid for the determined irradiation period.
This may be accomplished via computer or any other known methods. Indeed, the methods
and systems of the present invention contemplate the predetermination of any of the variables
such as TELEV, FLEV, thickness ratio, irradiation period, uniform fluid thickness, non-target
thickness, and/or hematocrit value in the buffy coat. Any or all of these predetermined
variables may be accessible by the user, e.g., available in tabular form, and, in a particular
embodiment of the present invention, stored or accessible in computer memory.
In order to assess the accuracy of the calculated amount of UVA energy predicted by
equations 2.0 and 2.4, an equal number of lymphocytes were suspended in clear phosphate
buffered saline and in a buffy coat suspension with 3.5% hematocrit. These two suspensions
were exposed to a UVA light in the presence of 100 ng/ml of 8-MOP. Controls were also
provided in which no 8-MOP was added to the suspensions. The degree of the injury to the
cells by this treatment at the same 8-MOP concentration is dependent on the UVA energy
i
dosage and can be measured by the cell viability.
The irradiation periods were calculated by equations 2.0 and 2.4 to deliver
approximately 1.4 Joule/cm2 of UVA energy to the lymphocytes in the fluids. Since the
phosphate buffered saline is transparent to UVA light, the irradiation period was calculated
based on the incident irradiance (equation 2.0). The irradiation period for the lymphocytes in
the buffy coat suspension was calculated by equations 2.0 and 2.4. The post-irradiation cell
viability of both samples was measured to compare the injury to the cells. The cell viability
of both samples were around 19% or less seven days after the irradiation while that of the
untreated control sample was around 85% or higher. This result shows that the lymphocytes
in the phosphate buffered saline and the buffy coat suspension received the same amount of
injury and resultant cell death. Indeed, the lymphocytes in both samples received the same
amount of UVA energy as calculated by each equation.
Equation 2.0 may preferably be used with any partially transparent solutions or
suspensions. It requires an accurate transmittance (T) measurement of a known thickness (D)
of the fluid, preferably under conditions where the materials in the fluid are homogeneous.
Equation 2.4, may be particularly applicable with fluids comprising red blood cells.
Referring to the associated Figures, in a specific embodiment of the invention, Figure
I depicts a extracorporeal photopheresis system 100 as an application of phototherapy
according to the present invention as applied to the treatment of leukocytes. See PCT
Application WO 97/36581. The phototherapy system 100 includes a photoactivatable drug,
8-MOP 110, a patient 120, a biological fluid extraction device 130 for extracting blood, a
centrifuge device 140 to separate out the buffy coat from the blood, a photoactivation device
150, a fluid (i.e., buffy coat) insertion device 160, and a blood insertion device 170. One
skilled in the art will appreciate that system 100 may contain additional or different devices
and can support a variety of phototherapy applications, as mentioned above. See U.S. Patent
Nos. 4,921,473, 4,838,852,5,147,289, 5,150,705, 5,383,847, 5,433,738, and 5,459,322, each
of which are expressly incorporated by reference herein and relate to various applications to
which the systems and apparatus of the present invention can be utilized.
Figures 2A and 2B depict a flow diagram 200 of the blood in the photopheresis
system in Figure 1. The first step is to mix the patient's 120 blood with 8-MOP 110 (step
202). In the present embodiment, the patient 120 is orally administered the 8-MOP 110 and,
over the course of a few hours, the drug mixes with the patient's 120 blood. Next, after the
drug 110 sufficiently interacts with the blood (step 204) an amount of blood-drug mixture is
extracted 130 (step 206) and transferred to a separator, such as a centrifuge device 140 (step
208).
After the blood-drug mixture is transferred to the centrifuge device 140, the centrifuge
device 140 separates the mixture (step 210). A particular centrifuge device uses an optical
sensor to determine when to separate (or skim) the fluid. First, the centrifuge skims off the
plasma, then the buffy coat, which contains the target material (i.e., 8-MOP in the
leukocytes), and then the red blood cells. The centrifuge device uses an optic sensor located
inside the centrifuge chamber that measures deflected light. This optic sensor, by measuring
the deflected light in the centrifuge determines when to skim off the separated fluids or
material. After separation, the buffy coat and a percentage of plasma are re-combined. The
plasma is the medium in which the leukocytes and 8-MOP reside. Even after separation,
however, the separated buffy coat and plasma mixture may comprise some red blood cells
and platelets, since the separation process may not be able to achieve complete separation.
These remaining red blood cells and platelets, contained in the buffy coat, are the non-target
attenuators of light. In the present embodiment, the red blood cells are the dominant non-
targets since they are the major attenuators of light, when compared to other attenuating
material in the target fluid.
Once the target fluid (i.e., the buffy coat mixture) is separated, a second optic sensor
determines whether the target fluid contains a desired hematocrit (percentage of red blood
cells) (step 212). In a particular embodiment, a desired hematocrit is about one (1) to two (2)
percent. This second optic sensor, which measures transmittance, determines whether a
desired hematocrit is reached (i.e., 1 % in the present embodiment). If the hematocrit
percentage is not at the desired percentage, then additional blood-drug mixture is processed
by the centrifuge (step 210).
If the non-target fluid contains the desired hematocrit percentage, then the centrifuge
determines what separated fluid it is processing (step 214). If the centrifuge is processing the
non-target fluid, then the centrifuge combines the remaining separated plasma with the
separated red blood cells and transfers the mixture to the separated blood insertion device 170
(step 216). Then, the blood insertion device returns the red blood cell/plasma mixture to the
patient (step 218) and processing stops.
If the centrifuge is processing the target fluid, the centrifuge then transfers the target
fluid to the photoactivation device (step 220). Step 220 and step 216 may happen
concurrently. The photoactivation chamber 150 then irradiates the fluid for a period of time
(step 222). Computer 300 controls the photoactivation chamber 150 as illustrated in Figure 3
and described in the corresponding discussion. The target fluid, now treated, is then
transferred to a fluid insertion device 160 (step 224). Then, the target insertion device returns
the red blood cell/plasma mixture to the patient (step 226) and processing stops.
Figure 3 is a diagram of a computer 300 for controlling the photoactivation device
150 according to the implementation of the present invention. The computer 300 includes a
memory 310, a central processing unit (CPU) 320, a photoactivation interface 330, an
operator interface 340, an input device 350, and a video display 360. One skilled in the art
will appreciate that computer 300 may contain additional or different components. The
memory 310 further includes an operating system 312, a photoactivation program 314, and
look-up table 315. The look-up table 315 may comprise a storage location in the memory
310 and may contain tables that correspond to data needed by the photoactivation program
314. The individual tables and the corresponding data are described in further detail in the
descriptions that correspond to Figures 4 through 9. The photoactivation program 312
acquires the FLEV. The FLEV could be obtained by accessing the look-up table 315, via the
input device 350, or by calculation as further described in the descriptions that correspond to
Figures 4 through 9.
Although aspects of the present invention are described as being stored in memory
310, one skilled in the art will appreciate that one or more of these aspects may also be stored
in other computer-readable media, such as secondary storage devices, like hard disks, floppy
disks, or CD-ROMs; a carrier wave from the Internet; or other forms of RAM or ROM.
Indeed, each of the methods, or particular steps contained therein, may be performed by or
stored in a computer or computer readable media.
Figure 4 depicts a flowchart 400 of the steps performed by the photoactivation
program 314 when requested to determine and then deliver an amount of light energy to a
fluid containing targets whereby the targets in the fluid will receive an effective amount of
light energy. The first step performed by the photoactivation program 314 is to obtain the
TELEV (step 402). The desired result is previously defined and is based on the phototherapy
application. For instance, when photopheresis is used to treat CTCL, the TELEV applied to
the leukocytes preferably causes at least fifty (50) percent of the leukocytes to gradually die
within six (6) days after exposure to the light energy.
The TELEV may be obtained by accessing, for example, a look-up table 315 that
contains TELEV data. In an alternative embodiment of the present invention, the
photoactivation program 314 may obtain the TELEV via the input device 350. Figure 5
illustrates how the TELEV may be clinically identified once the desired result is known.
Once the TELEV is obtained, the next step is to obtain the average light energy factor
for the fluid (step 404). The ALE factor is the percent of incident light energy that will be
delivered to an average unit area of fluid. The ALE factor may be obtained by accessing the
portion of the look-up table 315 that pertains to ALE factor data. In an alternative
embodiment of the present invention, the ALE factor may be obtained via the input device
350.
In an alternative embodiment of the present invention, the ALE factor may be
obtained for any target in a biological fluid from knowing the average light energy value
(Joules/cm2) at the unit surface area of the targets in the fluid and knowing the light energy
value (Joules/cm )-at the incident surface of the biological fluid. The description that
accompanies Figure 6 illustrates such a procedure for obtaining the ALE factor.
In an alternative embodiment of the present invention, the ALE factor may be
obtained from knowing the fluid's thickness ratio and the light transmittance value of a
known fluid thickness. The thickness ratio is the ratio of the uniform thickness of the fluid
and the average thickness of the non-target in the fluid. The non-target is material in the fluid
that attenuates light energy. The description that accompanies Figure 7 illustrates such a
procedure for obtaining the ALE factor.
In an alternative embodiment of the present invention, when fluid comprises red
blood cells as non-targets that attenuate light energy, the ALE factor may be obtained from
knowing the thickness ratio and knowing the percentage of hematocrit or red blood cells in
the fluid. The description that accompanies Figure 8 illustrates such a procedure for
obtaining the ALE factor.
In an alternative embodiment of the present invention, when fluid comprises red
blood cells as non-targets that attenuate light energy, the ALE factor may be obtained from
knowing the uniform thickness of the fluid and knowing the percentage of hematocrit or red
blood cells in the fluid. The description that accompanies Figure 9 illustrates such a
procedure for obtaining the ALE factor.
After obtaining the ALE factor, the next step is to obtain the FLEV or the amount of
light energy needed to be delivered to the fluid so that the targets in the fluid will receive the
TELEV (step 406). In a preferred embodiment, the FLEV can be calculated by knowing the
TELEV and the ALE factor and using equation 1.0, as described previously.
After obtaining the FLEV, one may then obtain the irradiation time period (step 408).
The irradiation time period is the amount of time needed for the lamp or light energy source
to deliver the FLEV to the fluid. The irradiation time period is obtained by accessing the
portion of the look-up table 315 that pertains to irradiation time period data.
In an alternative embodiment of the present invention, the irradiation time period can
be calculated. Factors that might be considered in irradiation time period calculation are
lamp decay or power, the shape of the lamp, or the volume of fluid to be irradiated. In an
alternative embodiment of the present invention, when the fluid comprises non-target red
blood cells, the irradiation time period can be calculated knowing the fluid's volume, the
percent of red-blood cells in the fluid, and the decay life of the light source using, for
example, an equation such as equation 1.5, as described previously.
After obtaining the irradiation time period, one may then instruct the photoactivation
device 150 to engage the light energy lamp for the irradiation time period.
Figure 5 depicts a flowchart 500 of the steps performed when clinically obtaining the
TELEV. The first step in clinically obtaining the TELEV is to obtain the desired result of the
phototherapy (step 502). The next step is to place sample targets in a non-attenuating fluid,
which is often a biological or chemical fluid (step 504). One skilled in the art will recognize
that there are numerous non-fluid mediums and other fluid types that can support targets such
as saline, and filtered plasma. In an alternative embodiment, when targets initially reside in a
fluid, samples of the fluid can be used for the clinical tests, provided any or most of the non-
attenuation materials are filtered out.
Next, samples of the fluid containing the targets are irradiated with varying amounts
of light energy (step 506). After irradiating the sample fluids, a TELEV is identified that
corresponds to the sample that produced the desired the result (step 508). One skilled in the
art will appreciate that any TELEV is specific to the particular application of the methods and
systems of the present invention.
Figure 6 depicts a flowchart 600 of the steps performed by the photoactivation
program 314 when obtaining the ALE factor. This procedure for obtaining the ALE factor
may be used for any fluid containing targets.. The first step to obtain the ALE factor is to
obtain the average light energy value at the unit surface area of the targets in the fluid (step
602). The average light energy value at the unit surface area can be obtained by accessing the
portion of the look-up table 315 that pertains to average light energy value at the unit surface
area data. In an alternative embodiment of the present invention, the photoactivation
program 314 may obtain the average light energy value at the unit surface area via the input
device 350.
The next step is to obtain the light energy value at the incident surface of the
biological fluid (step 604). The light energy value at the incident surface can be obtained by
accessing the portion of the look-up table 315 that pertains to light energy value at the
incident surface data. In an alternative embodiment of the present invention, the
photoactivation program 314 may obtain the light energy value at the incident surface via the
input device 350. The ALE factor may then be calculated using equation 1.0 (step 606).
Figure 7 depicts a flowchart 700 of the steps performed by the photoactivation
program 314 when obtaining the ALE factor. This procedure for obtaining the ALE factor
may be used for any biological fluid containing targets. However, the accuracy of this
equation is maximized when a homogeneous mixture of targets and non-targets in the fluid is
provided. In a particular embodiment of the present invention, a homogeneous biological
fluid mixture may be obtained by stirring the biological fluid containing the targets and non-
targets.
To obtain the ALE factor, one first obtains the thickness ratio of the fluid (step 702).
The thickness ratio is the ratio of the uniform thickness of the fluid and the average thickness
of the non-target in the fluid. The thickness ratio, the uniform fluid thickness, and the non-
target's thickness can be obtained by obtaining these values by, for example, accessing a
look-up table 315 that contains data relating to these parameters. In an alternative
embodiment of the present invention, the photoactivation program 314 may obtain the
thickness ratio, the uniform fluid thickness, and the non-target thickness via the input device
350. Once the uniform fluid thickness and the non-target thickness data are obtained, the
thickness ratio can be calculated by dividing the uniform fluid thickness by the non-target
thickness.
After obtaining the thickness ratio, one then may obtain a light transmittance value of
a known fluid thickness (step 704). The irradiation period can be obtained by accessing the
portion of a look-up table 315 that pertains to light transmittance value of a known fluid
thickness data. In an alternative embodiment of the present invention, the photoactivation
program 314 may obtain a light transmittance value of a known fluid thickness. The ALE
factor may then be calculated using equation 1.1 (step 706).
Figure 8 depicts a flowchart 800 of the steps performed by the photoactivation
program 314 when obtaining the ALE factor. This procedure for obtaining the ALE factor
may be used for biological fluid that comprises red blood cells as non-targets that attenuate
light energy. The accuracy of this equation may depend on how well the fluid is stirred. The
first step to obtain the ALE factor is to obtain the thickness ratio (step 802). The thickness
ratio is the ratio of the uniform thickness of the fluid and the average thickness of the non-
target in the fluid. The non-target is the material in the fluid that attenuates light energy. The
thickness ratio, the uniform fluid thickness, and the non-target's thickness can be obtained by
accessing the portion of the look-up table 315 that pertains to thickness ratio, the uniform
fluid thickness, and the non-target thickness data, respectively. In an alternative embodiment
jf the present invention, the photoactivation program 314 may obtain the thickness ratio, the
uniform fluid thickness, and the non-target's thickness via the input device 350. Once
obtaining the uniform fluid thickness and the non-target thickness data, the thickness ratio
can be calculated by dividing the uniform fluid thickness by the non-target thickness.
After obtaining the thickness ratio, the next step is to obtain percentage of red blood
cells or hematocrit per unit of biological fluid (step 804). The red-blood cell percentage can
be obtained by reading, for example, the optical or electromagnetic profile of the fluid by
known means or by accessing the portion of the look-up table 315 that pertains to red-blood
cell percentage data. In an alternative embodiment of the present invention, the
photoactivation program 314 may obtain the red-blood cell percentage via the input device
350. The ALE factor may then be calculated using equation 1.2 (step 806).
Figure 9 depicts a flowchart 900 of the steps performed by the photoactivation
program 314 when obtaining the ALE factor. This procedure for obtaining the ALE factor
may be used for biological fluid that comprises red blood cells as non-targets that attenuate
light energy and have a stacking factor of between 1 and 2. The accuracy of the results of
this equation may depend on how well the fluid is stirred. The first step to obtain the ALE
factor is to obtain the uniform fluid thickness (step 802). The uniform fluid thickness can be
obtained by accessing the portion of the look-up table 315 that pertains to uniform fluid
thickness data. In an alternative embodiment of the present invention, the photoactivation
program 314 may obtain the uniform fluid thickness via the input device 350.
After obtaining the uniform fluid thickness, the next step is to obtain the percentage of
red blood cells or hematocrit per unit of biological fluid (step 904). The red blood cell
percentage can be obtained by reading, for example, the optical or electromagnetic profile of
the fluid by known means or by accessing the portion of the look-up table 315 that pertains to
red blood cell percentage data. In an alternative embodiment of the present invention, the
photoactivation program 314 may obtain the red blood cell percentage via the input device
350. The ALE factor may then be calculated using equation 1.3 (step 906).
Figure 10 depicts a graph of ALE factors calculated for a fluid comprising red blood
cells as non-targets for three different fluid thicknesses (1 mm, 2 mm, and 3 mm). These
ALE factors were calculated using equations 1.1 (Analytical Model) and 1.3 (Stacking
Model). The ratio of the average light energy delivered to the targets in the fluid and the light
energy delivered to the incident point is plotted as a fenction: of percent hematocrit at
different fluid thickness.
The present invention is not to be limited in scope by the specific embodiments
described which are intended as single illustrations of individual aspects of the invention and
functionally equivalent methods and components are within the scope of the invention, in
addition to those shown and described herein will become apparent to those skilled in the art
from the foregoing description and accompanying drawings. Such modifications are intended
to fall within the scope of the appended claims.
We Claim:
1. A method of determining a fluid light energy value for delivery to
a biological fluid comprising targets and non-target material,
wherein an effective amount of light energy is desired to be
delivered to said targets and wherein said non-target material
attenuates said light energy destined for said targets, comprising
the steps of:
obtaining said target's effective light energy value;
obtaining said fluid's average light energy factor; and
calculating said fluid light energy value for delivery to said
biological fluid, wherein determining the fluid light energy value is
performed by one or more computer processors.
2. The method as claimed in claim 1 comprising the step of:
delivering the fluid light energy value to said biological fluid.
3. The method as claimed in claim 1, wherein determining the fluid's
light energy value is performed by one or more computer
processors.
4. The method as claimed in claim 1 comprising the step of:
providing a homogenous biological fluid mixture.
5. The method as claimed in claim 1, wherein said non-target
material comprises red blood cells.
6. The method as claimed in claim 5, wherein obtaining said average
light energy factor comprises the steps of:
obtaining a thickness ratio;
obtaining a red blood cell percentage for said biological fluid; and

calculating the average light energy factor for said targets in said
biological fluid.
7. The method as claimed in claim 6, wherein obtaining said average
light energy factor is performed by one or more computer
processors.
8. The method as claimed in claim 6, wherein obtaining said
thickness ratio comprises the step of:
accessing a thickness ratio table.
9. The method as claimed in claim 6, wherein obtaining said red
blood cell percentage comprises the step of:
accessing a red blood cell percentage table.
10. The method as claimed in claim 6 comprising the step of:
Providing a homogenous biological fluid mixture.
11. The method as claimed in claim 6, wherein said biological fluid
comprises leukocyte-rich buffy coat.
12. The method as claimed in claim 11, wherein said biological fluid is
treated with a light energy activatable drug.
13. The method as claimed in claim 12, wherein said drug comprises
8-methoxypsoralen.
14. The method as claimed in claim 13, wherein said light energy
comprises ultraviolet light energy.
15. The method as claimed in claim 14, wherein said ultraviolet light
energy comprises ultraviolet A light energy.
16. The method as claimed in claim 15, wherein determining the fluid
light energy value is performed by one or more computer
processors.

17. The method as claimed in claim 13, wherein obtaining said red
blood cell percentage comprises the step of:
accessing a red blood cell percentage table.
18. The method as claimed in claim 6, wherein obtaining said
thickness ratio comprises the steps of:
obtaining a uniform thickness for said biological fluid;
obtaining a thickness for said non-target; and
calculating the thickness ratio.
19. The method as claimed in claim 18, wherein obtaining said
thickness ratio is performed by one or more computer processors.
20. The method as claimed in claim 18, wherein obtaining said
uniform film thickness comprises the step of:
accessing an uniform thickness ratio table.
21. The method as claimed in claim 18, wherein obtainmg said
thickness for said non-target comprises the step of:
accessing a non-target thickness table.
22. The method as claimed in claim 5, wherein obtainmg said average
light energy factor comprises the steps of:
obtaining a uniform thickness for said biological fluid;
obtaining a red blood ceil percentage for said biological fluid; and
calculating the average light energy factor for said targets in said
biological fluid.
23. The method as claimed in claim 22, wherein theoretical stacking of
said red-blood cells does not occur.
24. The method as claimed in claim 22, wherein said biological fluid
comprises up to about twenty percent said red blood cells.


25. The method as claimed in claim 22 comprising the step of:
obtaining a stacking factor.
26. The method as claimed in claim 22, wherein said stacking factor is
between 1 and 2.
27. The method as claimed in claim 26, wherein said stacking factor is
1.5.
28. The method as claimed in claim 22, wherein obtaining said
average light energy factor is performed by one or more computer
processors.
29. The method as claimed in claim 22, wherein obtaining said
uniform thickness comprises the step of:
accessing a biological fluid thickness table.
30. The method as claimed in claim 22, comprising the step of:
providing a homogenous biological fluid mixture.
31. The method as claimed in claim 22, wherein said biological fluid
comprises leukocyte-rich buffy coat.
32. The method as claimed in claim 31, wherein said biological fluid is
treated with a light energy activatable drug.
33. The method as claimed in claim 32, wherein said drug comprises
8-methoxypsoralen.
34. The method as claimed in claim 33, wherein said light energy
comprises ultraviolet light energy.
35. The method as claimed in claim 34, when said ultraviolet light
energy comprises ultraviolet A light energy.
36. The method as claimed in claim 35, wherein determining said fluid
light energy value is performed by one or more computer
processors.

37. The method as claimed in claim 1, wherein said biological fluid
comprises leukocyte-rich buffy coat.
38. The method as claimed in claim 37, wherein said biological fluid is
treated with a light energy activatable drug.
39. The method as claimed in claim 38, wherein said drug comprises
8-methoxypsoraien.
40. The method as claimed in claim 39, wherein said light energy
comprises ultraviolet light energy.
41. The method as claimed in claim 40, wherein said ultraviolet light
energy comprises ultraviolet A light energy.
42. The method as claimed in claim 41, wherein determining said fluid
light energy value is performed by one or more computer
processors.
43. The method as claimed in claim 1, wherein said calculating step
comprises:
calculating an irradiation time period required by a light energy
source to deliver said fluid's light energy value.
44. The method as claimed in claim 43 comprising the step of:
delivering the fluid's light energy value to said biological fluid.
45. The method as claimed in claim 43, wherein calculating the
irradiation time period is performed by one or more computer
processors.
46. The method as claimed in claim 43, wherein calculating the
irradiation time period comprises:
obtaining a decay life value for said light energy source.


47. The method as claimed in claim 43, wherein said non-
material comprises red blood cells, and wherein calculating the
irradiation time period comprises:
obtaining a volume of bio logical fluid value;
obtaining a percent of red blood cells value; and
obtaining a decay life value for said light energy source.
48. The method as claimed in claim 1, wherein obtaining said targets'
effective light energy value comprises the step of:
accessing a targets' effective light energy value table.
49. The method as claimed in claim 48, wherein obtaining said target's
effective light energy value is performed by one or more computer
processors.
50. The method as claimed in claim 1, wherein obtaining said target's
effective light energy value comprises the steps of:
placing said targets in fluid, wherein said targets in said fluid do
not receive attenuated light energy; and
irradiating said fluid with sample light energy values.
51. The method as claimed in claim 50, wherein said fluid comprises
saline.
52. The method as claimed in claim 50, wherein said fluid comprises
plasma.
53. The method as claimed in claim 1, wherein obtaining said target's
effective light energy value comprises the steps of:
placing said leukocytes in fluid comprises saline; and
identifying a light energy value whereby a desired percentage of
said leukocytes will die over the course of a specified time after
exposure to said light energy.

54. The method as claimed in claim 1, wherein obtaining said target's
effective light energy value comprises the steps of:
obtaining sample biological fluids from donors;
irradiating said targets in said biological fluids with sample light
energy values; and
determining the target's effective light energy value.
55. The method as claimed in claim 1, wherein obtaining said average
light energy factor comprises the step of:
accessing a light energy factor table.
56. The method as claimed in claim 55, wherein obtaining said
average light energy factor is performed by one or more computer
processors.
57. The method as claimed in claim 1, wherein the obtaining said
average light energy factor comprises the steps of:
obtaining an average light energy value at a unit surface area of the
targets in the biological fluid;
obtaining a light energy value at an incident surface of the
biological fluid film; and
calculating the average light energy factor for said targets in said
biological fluid.
58. The method as claimed in claim 57, wherein obtaining said
average light energy factor is performed by one or more computer
processors.
59. The method as claimed in claim 57, wherein obtaining said
average light energy value at a unit surface area comprises the step
of:
accessing an average light energy value at a unit surface area table.

60. The method as claimed in claim 57, wherein obtaining said light
energy value at an incident surface comprises the step of:
accessing a light energy value at an incident surface table.
61. The method as claimed in claim 57, comprising the step of:
providing a homogenous biological fluid mixture.
62. The method as claimed in claim 28, wherein said non-target
material comprises red blood cells.
63. The method as claimed in claim 57, wherein said biological fluid
comprises leukocyte-rich buffy coat.
64. The method as claimed in claim 63, wherein said biological fluid is
treated with a light energy activatable drug.
65. The method as claimed in claim 64, wherein said drug comprises
8-methoxypsoralen.
66. The method as claimed in claim 65, wherein said light energy
comprises ultraviolet light energy.
67. The method as claimed in claim 66, wherein said ultraviolet light
energy comprises ultraviolet A light energy.
68. The method as claimed in claim 67, wherein determining said fluid
light energy value is performed by one or more computer
processors.
69. The method as claimed in claim 1, wherein obtaining said average
light energy factor comprises the steps of:
obtaining a thickness ratio;
obtaining a light transmittance value of a known fluid film
thickness; and
calculating the average light energy factor for said targets in said
biological fluid.

70. The method as claimed in claim 69, wherein obtaining said
average light energy factor is performed by one or more computer
processors.
71. The method as claimed in claim 69, wherein obtaining said
thickness ratio comprises the step of:
accessing a thickness ratio table.
72. The method as claimed in claim 69, wherein obtaining said
irradiation period comprises the step of:
accessing a irradiation period table.
73. The method as claimed in claim 69, comprising the step of:
providing a homogenous biological fluid mixture.
74. The method as claimed in claim 69, wherein said non-target
material comprises red blood cells.
75. The method as claimed in claim 42, wherein obtaining said
thickness ratio comprises the steps of:
obtaining a uniform thickness for said biological fluid;
obtaining a thickness for said non-target; and
calculating the thickness ratio.
76. The method as claimed in 75, wherem obtaining said uniform film
thickness comprises the step of:
accessing a uniform thickness table.
77. The method as claimed in claim 75, wherein obtaining said non-
target thickness comprises the step of:
accessing a non-target thickness table.
78. The method as claimed in claim 75, wherein said non-targets
comprise red blood cells.

79. The method as claimed in claim 78, wherein obtaining said
thickness ratio is performed by one or more computer processors.
80. The method as claimed in claim 69, wherein said biological fluid
comprises the leukocyte-rich buffy coat.
81. The method as claimed in claim 80, wherein said biological fluid is
treated with a light energy activatable drug.
82. The method as claimed in claim 81, wherein said drug comprises
8-methoxypsoralen.
83. The method as claimed in claim 82, wherein said light energy
comprises ultraviolet light energy.
84. The method as claimed in claim 83, wherein said ultraviolet light
energy comprises ultraviolet A light energy.
85. The method as claimed in claim 84, wherein determining the
desired amount of said ultraviolet light energy is performed by one
or more computer processors.
86. A computer system for determining a fluid light energy value for
delivery to a biological fluid comprising targets and non-target
material, wherein an effective amount of light energy is desired to
be delivered to said targets and wherein said non-target material
attenuates said light energy destined for said targets, comprising:
a computer processor;
a memory which is operatively coupled to the computer processor;
and
a computer process stored in said memory which executes in the
computer processor and which includes:
an obtainer configured to obtain said target's effective light energy
value;

an obtainer configured to obtain said fluid's average light energy
factor; and
a calculator configured to calculate said fluid's light energy value
for delivery to said biological fluid.
87. The computer system as claimed in claim 86, wherein said non-
target material comprises red blood cells.
88. The computer system as claimed in claim 87, wherein said obtainer
configured to obtain the average light energy factor comprises the
steps of:
an obtainer configured to obtain a uniform thickness for said
biological fluid;
an obtainer configured to obtain a red blood cell percentage for
said biological fluid; and
a calculator configured to calculate the average light energy factor
for said targets in said biological fluid.
89. The computer system as claimed in claim 88, wherein theoretical
stacking of said red-blood cells does not occur.
90. The computer system as claimed in claim 88, wherein said
biological fluid comprises up to about twenty percent said red
blood ceils.
91. The computer system as claimed in claim 88, includes:
an obtainer configured to obtain said stacking factor.
92. The computer system as claimed in claim 88, wherein said
stacking factor is between 1 and 2.
93. The computer system as claimed in claim 92, wherein said
stacking factor is 1.5.

94. The computer system as claimed in claim 88, wherein said obtainer
configured to obtain said uniform thickness includes:
an accessor configured to access a biological fluid thickness table.
95. The computer system as claimed in claim 88, wherein said obtainer
configured to obtain said red blood cell percentage includes:
an accessor configured to access a red blood cell percentage table.
96. The computer system as claimed in claim 88, wherein said
biological fluid comprises leukocyte-rich buffy coat.
97. The computer system as claimed in claim 96, wherein said
biological fluid is treated with a light energy activatable drug.
98. The computer system as claimed in claim 97, wherein said drug
comprises 8-methoxypsoralen.
99. The computer system as claimed in claim 98, wherein said light
energy comprises ultraviolet light energy.
100. The computer system as claimed in claim 99, wherein said
ultraviolet light energy comprises ultraviolet A light energy.
101. The computer system as claimed in claim 86, wherein said
biological fluid comprises leukocyte-rich buffy coat.
102. The computer system as claimed in claim 101, wherein said
biological fluid is treated with a light energy activatable drug.
103. The computer system as claimed in claim 102, wherein said drug
comprises 8-methoxypsoraien.
104. The computer system as claimed in claim 103, wherein said light
energy comprises ultraviolet light energy.
105. The computer system as claimed in claim 104, wherein said
ultraviolet light energy comprises ultraviolet A light energy.


106. The computer system as claimed in claim 86, wherein said
calculator configured to calculate said fluid's light energy value for
delivery to said biological fluid includes:
A calculator configured to calculate an irradiation period required
by a light energy source to deliver said fluid's tight energy value.
107. The computer system as claimed in claim 106, wherein said
calculator configured to calculate said fluid's light energy value for
delivery to said biological fluid includes:
an obtainer to obtain a decay life value for said light energy source.
108. The computer system as claimed in claim 106, wherein said
biological fluid comprises non-target material, wherein said non-
target material comprises red blood cells, and wherein said
calculator configured to calculate said fluid's light energy value for
delivery to said biological fluid includes:
an obtainer to obtain a volume of bio logical fluid value;
an obtainer to obtain a percent of red blood cells value; and
an obtainer to obtain a decay life value for said light energy source.
109. The computer system as claimed in claim 86, wherein said obtainer
configured to obtain said target's effective light energy value
includes:
an accessor configured to access a targets effective light energy
value table.
110. The computer system as claimed in claim 86, wherein said obtainer
configured to obtain an average light energy factor includes:
an accessor configured to access a light energy factor table.


111. The computer system as claimed in claim 110, wherein said obtainer
configured to obtain said average light energy value at a unit surface
area includes:
an accessor configured to access an average light energy value at a
unit surface area table.
112. The computer system as claimed in claim 111, wherein said obtainer
configured to obtain said light energy value at an incident surface
includes:
an accessor configured to access a light energy value at an incident
surface table.
113. The computer system as claimed in claim 112, wherein said non-
target material comprises red blood cells.
114. The computer system as claimed in claim 86, wherein said obtainer
configured to obtain an average light energy factor includes:
an obtainer configured to obtain an average light energy value at a
unit surface area of the targets in the biological fluid;
an obtainer configured to obtain a light energy value at an incident
surface of the biological fluid film; and
a calculator configured to calculate the average light energy factor for
said biological fluid.
115. The computer system as claimed in claim 114, wherein said biological
fluid comprises leukocyte-rich buffy coat
116. The computer system as claimed in claim 115, wherein said biological
fluid is treated with a fight energy activatable drug.
117. The computer system as claimed in claim 116, wherein said drug
comprises 8-methoxypsoralen.

118. The computer system as claimed in claim 117, wherem said light
energy comprises ultraviolet light energy.
119. The computer system as claimed in claim 118, wherein said
ultraviolet light energy comprises ultraviolet a light energy.
120. The computer system as claimed in claim 86, wherem said obtainer
configured to obtain an average light energy factor includes:
An obtainer configured to obtain a thickness ratio;
An obtainer configured to obtain a light transmittance value of a
known fluid film thickness; and
A calculator configured to calculate the average light energy factor for
said biological fluid
121. The computer system as claimed in claim 120, wherein said obtainer
configured to obtain the thickness ratio further includes:
An accessor configured to access a thickness ratio table.
122. The computer system as claimed in claim 120, wherein said obtainer
configured to obtain an irradiation period includes:
an accessor configured to access a irradiation period table.
123. The computer system as claimed in claim 120, wherein said non-
target material comprises red blood cells.
124. The computer system as claimed in claim 120, wherem said biological
fluid comprises leukocyte-rich buffy coat
125. The computer system as claimed in claim 124, wherein said biological
fluid is treated with a light energy activatable drug.
126. The computer system as claimed in claim 125, wherein said drug
comprises 8-methoxypsoralen.

127. The computer system as claimed in claim 126, wherein said light
energy comprises ultraviolet light energy.
128. The computer system as claimed in claim 127, wherein said
ultraviolet light energy comprises ultraviolet a light energy.
129. The computer system as claimed in claim 120, wherein said obtainer
configured to obtain the thickness ratio includes:
an obtainer configured to obtain a uniform thickness for said
biological fluid;
an obtainer configured to obtain a thickness for said non-target; and
a calculator configured to calculate the thickness ratio.
130. The computer system as claimed in claim 129, wherein said obtainer
configured to obtain uniform film thickness includes:
an accessor configured to access a uniform thickness table.
131. The computer system as claimed in claim 129, wherein said obtainer
configured to obtain said non-target thickness includes:
an accessor configured to access a non-target thickness table.
132. The computer system as claimed in claim 129, wherein said non-
targets comprise red blood cells.
133. The computer system as claimed in claim 87, wherein said obtainer
configured to obtain the average light energy factor includes:
an obtainer configured to obtain a thickness ratio;
an obtainer configured to obtain a red blood cell percentage for said
biological fluid; and
a calculator configured to calculate the average light energy factor for
said targets in said biological fluid,

134. The computer system as claimed in claim 133, wherein said obtainer
configured to obtain the thickness ratio includes:
an accessor configured to access a thickness ratio table.
135. The computer system as claimed in claim 133, wherein the obtainer
configured to obtain the red blood cell percentage includes:
an accessor configured to access a red blood cell percentage table.
136. The computer system as claimed in claim 133, wherem said biological
fluid comprises leukocyte-rich buffy coat
137. The computer system as claimed in claim 136, wherein said biological
fluid is treated with a light energy activatable drug.
138. The computer system as claimed in claim 137, wherein said drug
comprises 8-methoxypsoralen.
139. The computer system as claimed in claim 138, wherein said light
energy comprises ultraviolet light energy.
140. The computer system as claimed in claim 139, wherein said
ultraviolet light energy comprises ultraviolet a light energy.
141. The computer system as claimed in claim 133, wherein said obtainer
configured to obtain the thickness ratio includes:
an obtainer configured to obtain a uniform thickness for said
biological fluid;
an obtainer configured to obtain a thickness for said non-target; and
a calculator configured to calculate the thickness ratio.
142. The computer system as claimed in claim 141, wherein said obtainer
configured to obtain me uniform film thickness includes:
an accessor configured to access an uniform thickness ratio table.


143. The computer system as claimed in claim 141, wherein said obtainer
configured to obtain the thickness for said non-target comprises the
step of;
an accessor configured to access a non-target thickness table,
144. A system for determining a fluid light energy value for delivery to a
biological fluid comprising targets and non-target material, wherein
an effective amount of light energy is desired to be delivered to said
targets and wherem said non-target material attenuates said light
energy passing through said biological fluid, comprising :
means for obtaining said target's effective light energy value;
means for obtaining an average light energy factor for said biological
fluid; and
means for calculating said fluid tight energy value for delivery to said
biological fluid, wherein determining the fluid light energy value is
performed by one or more computer processors.
145. The system as claimed in claim 144, wherein obtaining said targets'
effective light energy value comprises :
means for placing said targets in fluid, wherein said targets in said
fluid do not receive attenuated light energy; and
means for irradiating said fluid with sample light energy values.
146. The system as claimed in claim 144, wherein the obtaining said
average light energy factor comprises :
means for obtaining the average light energy value at a unit surface
area of the targets in the biological fluid;
means for obtaining the light energy value at an incident surface of the
biological fluid film; and

means for calculating the average light energy factor said targets in
said biological fluid.
147. The system as claimed in claim 144, wherein obtaining said average
light energy factor comprises;
means for obtaining a thickness ratio;
means for obtaining a light transmittance value of a known fluid film
thickness; and
means for calculating the average light energy factor for said targets
in said biological fluid.
148. The system as claimed in claim 147, wherein obtaining said thickness
ratio comprises:
means for obtaining a uniform thickness for said biological fluid;
means for obtaining a thickness for said non-target; and
means for calculating the thickness ratio.
149. The system as claimed in claim 144, wherein said non-target material
comprises red blood cells and wherein obtaining the average light
energy factor comprises;
means for obtaining a thickness ratio;
means for obtaining a red blood cell percentage for said biological
fluid; and
means for calculating the average light energy factor for said targets
in said biological fluid
150. The system as claimed in claim 149, wherein obtaining said thickness
ratio comprises:
means for obtaining a uniform thickness for said biological fluid;

means for obtaining a thickness for said non-target; and
means for calculating the thickness ratio.
151. The system as claimed 144, wherein said non-target material
comprises red blood cells and wherein obtaining the average light
energy factor comprises:
means for obtaining an uniform thickness for said biological fluid;
means for obtaining a red blood cell percentage for said biological
fluid; and
means for calculating the average light energy factor for said targets
insaid biological fluid.

Documents:

383-CAL-2000 -(22-12-2011)-FORM-27.pdf

383-cal-2000-assignment.pdf

383-cal-2000-correspondence.pdf

383-cal-2000-examination report.pdf

383-cal-2000-form 18.pdf

383-cal-2000-form 26.pdf

383-cal-2000-form 3.pdf

383-cal-2000-form 5.pdf

383-CAL-2000-FORM-27.pdf

383-cal-2000-granted-abstract.pdf

383-cal-2000-granted-claims.pdf

383-cal-2000-granted-description (complete).pdf

383-cal-2000-granted-drawings.pdf

383-cal-2000-granted-form 1.pdf

383-cal-2000-granted-form 2.pdf

383-cal-2000-granted-specification.pdf

383-cal-2000-others.pdf

383-cal-2000-reply to examination report.pdf

383-cal-2000-translated copy of priority document.pdf


Patent Number 246493
Indian Patent Application Number 383/CAL/2000
PG Journal Number 09/2011
Publication Date 04-Mar-2011
Grant Date 01-Mar-2011
Date of Filing 06-Jul-2000
Name of Patentee THERAKOS, INC.
Applicant Address 437 CREAMARY WAY, EXTON, PENNSYLVANIA
Inventors:
# Inventor's Name Inventor's Address
1 LEE KYU HO 605 CORNERSTONE LANE, BRYN MAWR, PA 19010
PCT International Classification Number A61M 1/36
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 09/350 134 1999-07-09 U.S.A.