Title of Invention

AN APPARATUS FOR DISINFECTING BIOLOGICAL MATERIALS

Abstract

A novel apparatus for disinfecting objects using chemical steriliant solutions, made of material that can be steam sterilized and consisting of cylindrical or conical vessel with a wide mouth, closed with a watertight, hinged snap-fit lid, provided with a side drainage port with a similar closure, a perforated diaphragm to prevent floating away of the objects when draining solutions and modifications of the apparatus such as a double walled vessel with the lower part of inner chamber being perforated and a further modification that enables the lower perforated cup shaped part to be removed and replaced with a component of different perforation size, an additional access port on the upper lid to enable connection of flexible tubing, perforated basket shaped receptacle that fits into the inside of the apparatus and provided with vertical partitions so as to permit separation of objects placed therein and prevent adhesion between such objects, and method of using the said apparatus consisting of placing the objects in the inner chamber or in the perforated receptacle placed in the inner chamber, pouring the desired chemical solution into the chamber until they are submerged and swirling or shaking the apparatus to improve the contact of the objects with the chemical solution and after for a predetermined duration of the treatment, draining out the solutions through the side port by tilting the apparatus:

Full Text

Field of invention This invention in general relates to the field of biology/biotechnology. Specifically this invention relates to a process of disinfection/surface sterilization of biological materials using chemical steriliant solutions. More specifically the present invention relates to an apparatus that facilitates the surface sterilization of objects used in biological procedures with chemical sterilant solutions and rinsing with sterile water Background of the Invention and Prior Art The presence of microorganisms in the air and on any exposed surface is a well-known fact. Removal of the microbes from the surfaces of objects is done when there is a risk of their proliferation to a level where they cause harm or damage. Common examples are the spoilage of organic matter like leather, foodstuff and infection of wounds in human or animals from microbes present in the surroundings. Several methods are known which rid surfaces of microbes. Application of high temperatures, boiling, autoclaving and treatment with chemical substances are common enough methods in day-to-day life, which result in a good level of sterilization. Some procedures however require very high levels of sterility like surgery and use of prosthetic devices. Others involve use of living tissues and therefore preclude use of high temperature for killing of microbes. In vitro culture or tissue culture of plant or animal parts, isolation of pure cultures of microbes etc. are some of the applications of this nature. The need for ensuring that excised parts (explants) taken from plants are free of microbial organisms has been adequately described in literature ( George, 1993; Leifert and Waites, 1990) . The common methods used to achieve this are also described in almost every paper describing a tissue culture procedure. Literature dealing with microbiological techniques also descrit>es such methods. Bhefly the typical method of sterilizing plant tissues involves placing the material in a pre-sterilized container and pouring sufficient sterilant solution to cover it completely. After the required duration of treatment during which the container is gently agitated, the solution is drained out followed by rinsing of the explants with 2-5 changes of sterile water. The inherent difficulty when using plant parts or small bits of tissue is that because of the buoyancy of certain tissues they float away with the solution when it is drained out. Very small objects like seeds of some species and fern spores cannot be treated without placing them first in a doth or paper receptacle and then treating them as in the method described above. Yet another problem encountered with plant material like leaves is the tendency to stick to each other and prevent proper contact of all surfaces with the sterilant The normal method involves the use of flasks and bottles and the removal of the closures several times for introduction and removal of objects and solutions increases the chance of contaminating organisms gaining entry into the container during the different stages. A certain amount of skill on the part of the operator is therefore needed to avoid loss of explants and improper sterilization in all the situations described here. This contributes to the drudgery and time needed to carry out the procedures. This invention seeks to improve the procedure and eliminate some of the risks through the use of a novel apparatus. Object of the invention It is the primary object of this invention to invent an apparatus for use in the field of biology and biotechnology. It is another object of this invention to invent a process of disinfection/surface sterilization of biological materials using chemical steriliant solutions. it is another object of this invention to invent an apparatus that facilitates the surface sterilization of objects used in biological procedures with chemical sterilant solutions and rinsing with sterile water. It is another object of this invention to invent an apparatus that rectifies the disadvantages and simplifies the disinfecting procedures known to the art. It is another object of this invention to invent an apparatus that is made of autoclavable material and permits the treatment of objects with solutions of predetermined concentrations for a predetermined duration followed by draining away of the solutions without the risk of the loss of the objects and without compromising sterility. Further objects of the invention will be apparent from the ensuing description. STATEIWENT OF THE INVENTION A novel Apparatus for facilitating the surface sterilization of objects required for biological procedures, by treatment with chemical sterilant solutions, followed by rinsing with sterile water or fluids consisting of a cylindrical or conical vessel opening to the outside by a wide mouth and provided with a watertight, hinged, snap-fit lid on the top and a narrow side drainage port incorporating a perforated diaphragm and aiso provided with a watertight hinged snap- fit closure, wherein the lid is provided with a small upper access port with a removable closure and a sen^ated tube end that enables connection of flexible tubing as and when the application demands. Detailed description Revealed here is an Apparatus made of autoclavable material for facilitating the surface sterilization of objects and consisting of a chamber in which the objects placed therein to be sterilized are brought in contact with steriiant solutions and is provided with a hinged snap-fit lid on the top and a means of draining away the solutions through a side drainage port, fitted with a perforated diaphragm to retain the said objects within the chamber and with a hinged snap fit closure to facilitate opening and closing . A narrow access port with a removable closure and a serrated end is also provided on the lid to enable connection vwth flexible tubing. In a variation of the apparatus, the vessel is modified to form an inner chamber by insertion of an inner vessel of same shape but of smaller size, with the lower half of the said inner vessel (chamber) being perforated. In a further modification of the apparatus, the lower perforated part of the inner vessel is made as a removable cup shaped component so as to enable the replacement with a similar component of a different perforation size, chosen depending on the size of the objects to t>e sterilized. Yet another modification of the apparatus embodies the use of a perforated bolder with vertical compartments that fits inside the inner chamber and can be used to segregate or prevent adhesion of the objects to each other. Use of the said apparatus for surface disinfection is by treatment of the objects for predetermined duration with suitable chemical sterilant solutions follovwed by rinsing with sterile water. Depending on the type of object or objects to be sterilized, the perforated holder or the replaceable cup is chosen for the sterilizing procedure. The objects are placed in the inner chamber along with the sterilant solution through the lid at the top under sterile conditions. After the desired treatment duration, the solution is drained through the side drainage port by opening the snap^t closure with the fingers and tilting the apparatus. Sterile water used to rinse the objects free of the steriliant solution is added either by opening the lid itself or through the side drainage port or the port on the top of the lid and drained after the treatment as mentioned above. If the use of vacuum is desired within the vessel during the treatment, the narrow port on the lid is connected to the vacuum line using a tubing having an on-line sterile air filter. The nature of the apparatus and the method for its use will be apparent in the following description and explanatory diagrams. Figure 1, 2 and 3 shows the details of the arrangement of the different components of the invention. Figure 1 shows the scheme of the simpler form of the apparatus (Basic design) consisting of a cylindrical or conical container with a wide mouth provided with a watertight hinged snap-fit lid. The side drainage port is provided with a similar snap-fit hinged lid and also incorporates a perforated diaphragm that permits draining away of solutions while retaining the objects within. The lid is provided with a small upper access port with a removable closure and a serrated tube end that enables connection of flexible tubing as and when the application demands. A variation of the basic apparatus (the double-walled apparatus) is shown in Figure 2 wherein the vessel is modified by insertion of an inner vessel of the same shape but smaller size and a rim that forms the lid of the outer vessel, thereby forming an inner chamber circumscribed by a narrow outer chamber all around except for the upper extremity where it projects out to form a neck and a mouth covered with a watertight hinged snap- fit lid. The inner chamber is further provided with perforations on the lower extremity through which it is connected to the outer chamber. A further modification of the inner vessel is in the perforated part that is made so as to form a removable cup that can be replaced with a similar component of a different perforation size chosen depending on the size of the objects used for sterilization. The inner vessel rests on supports provided at the bottom of the cup so that it remains in place while the apparatus is in use. References 1. George, E.F. (1993) Controlling Persistent Contaminants and Plant Diseases, In: Plant Propagation by Tissue Culture, Part I: The Technology, Exegetics Limited, England, p. 130-162. 2. George, E.F. (1993) Equipment and procedures, In: Plant Propagation by Tissue Culture, Part I: The Technology, Exegetics Limited, England, p. 95-129 3. Leifert, C. and Waites, W.M. (1990) Contaminants of plant tissue cultures. Newsletter of International Assoc, of Plant Tiss. Cult. No 60:2-13. We claim: 1. A novel Apparatus for facilitating the surface sterilization of objects required for biological procedures, by treatment with chemical sterilant solutions, followed by rinsing with sterile water or fluids consisting of a cylindrical or conical vessel opening to the outside by a wide mouth and provided with a watertight, hinged, snap-fit lid on the top and a narrow side drainage port incorporating a perforated diaphragm and also provided with a watertight hinged snap- fit closure, wherein the lid is provided with a small upper access port with a removable closure and a serrated tube end that enables connection of flexible tubing as and when the application demands. 2. A novel Apparatus as in Claim 1 herein the vessel is modified to include a inner vessel of the same shape but of a smaller size, with a rim that forms the lid of the outer vessel and permitting insertion into the said outer vessel and leaving a narrow gap between the two walls all around the inner vessel and below, the bottom and lower half of the said inner vessel being provided with a plurality of apertures of a predetermined size preferably in the range 0.01 mm to 6 mm and chosen depending on the size of the objects to be used with the apparatus. 3. Apparatus as in Claim 1 in which the perforated lower part of the inner chamber is modified to enable a perforated cup shaped component that can be removed and replaced as desired with a similar component of appropriate perforation size depending on the size of the object to be sterilized. 4. A device to be used in conjunction with the apparatus as claimed in claim 1, consisting of a perforated basket shaped holder of a size permitting insertion into the inner chamber of the said apparatus and provided with multiple vertical partitions, also perforated, enabling the separation or preventing adhesion of objects placed in the compartments formed therein. Apparatus in Claim 1 in which the material used for manufacture is selected from a group consisting of, but not restricted to autodavable plastics like polypropylene, polycarbonate, polymethylpentane and polysulfone, so that the apparatus permits viewing of the objects placed in the inner chamber and also permits the apparatus to be rendered sterile before use by autoclaving. A modification of the lid of the apparatus as claimed in 1 and 2 wherein an additional narrow access port is provided having a serrated tube end to enable connection of flexible tubing and which is kept sealed by a elastic cap when not in use. A method of utilizing the apparatus in Claim 1. for disinfecting objects required to be substantially free of undesirable organisms for use in biological procedures requiring sterility, with a chemical solution having microbicidal properties, selected from a group consisting of, but not limited to, mercuric chloride, sodium hypochlorite, calcium hypochlorite, hydrogen peroxide and chlorine v^ter, at a concentration determined empirically based on the nature of the objects so that substantial damage to the said objects does not occur and substantial disinfection is achieved as ascertained by the absence of microbial contamination or damage to the objects after a reasonable period of time subsequent to the disinfection procedure and the application to which the object is put to. Method in claim 1 . where the object to be disinfected is placed, under substantially sterile conditions, in the chamber of the apparatus in Claim 1 . or in the perforated holder as claimed in Claim 4.., after the said apparatus has been autoclaved for 15-20 minutes at 121°C, is treated with a sterilizing chemical solution \Atiich Is poured into the inner chamber, retained for a predetermined duration, after which the said solution is drained out through the side port by opening the lid on the side drainage port and the said objects are rinsed several times by washing with sterile water poured into the inner chamber through the side port or the upper port and drained through the side port, and the disinfected objects are removed for further processing or application through the upper port under substantially sterile conditions. Method in Claim 2. where the treatment duration is determined by empirical tests on the object to be disinfected so that substantial damage to the said objects does not occur and substantial disinfection is achieved as ascertained by the absence of microbial contamination or damage to the objects after a reasonable period of time subsequent to the disinfection procedure and in the course of the application to which the object is put to. Method in Claim 1 & 2 further modified when trapped air bubbles are required to be removed from the objects to be disinfected, wherein the nan'ow upper port is connected, through a sterile gaseous filter, to a vacuum source for the duration of the treatment or part of it. A novel Apparatus and method for facilitating the surface sterilization of objects required for biological procedures as substantially described herein.

Documents:

1033-CHE-2003 AMANDED CLAIMS 10-03-2010.pdf

1033-CHE-2003 AMANDED PAGES OF SPECIFICATION 10-03-2010.pdf

1033-CHE-2003 CORRESPONDENCE OTHERS 10-03-2010.pdf

1033-che-2003-abstract.pdf

1033-che-2003-claims.pdf

1033-che-2003-correspondnece-others.pdf

1033-che-2003-description(complete).pdf

1033-che-2003-drawings.pdf

1033-che-2003-form 1.pdf

1033-che-2003-form 19.pdf

1033-che-2003-form 26.pdf