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MINUCELLS AND MINUTISSUE VERTRIEBS GMBH

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PAGE 14

Instructions for use part 3:


 

Starting the perf(i)usion culture: The sterilized working line is placed under the laminar air flow for the following procedures (15). The screw cap of the left glass bottle (16) is opened to fill in the prepared culture medium. The bottle is closed by tightening the screw cap. The re-usable autoclaved filters (17) are fixed at the top of both screw caps to ventilate the bottles. The lid of the culture container is opened (18) and the tissue carriers are placed into the culture container with sterile forceps (19). With a pipette the container is filled with culture medium and the lid of the container is closed and clamped down. The silicone tube is closed with a plastic clip during transport to avoid uncontrolled exchange of the medium. Then the complete working line is taken out of the laminar air flow and placed on a table in the unsterile atmosphere of the laboratory.


Per(i)fusion culture: For a per(i)fusion culture experiment a peristaltic pump and a thermo plate are necessary. The pump (20) is known from column chromatography equipment. The thermo plate (21) before used to stretch paraffin sections is suitable to keep the culture container at 37°C. If desired, the medium bottles can be placed into a cooled circulating water bath or in a bottle cooler with horizontal lids (22). After installing the working line in the peristaltic pump and placing the culture container on the thermo plate the per(i)fusion of culture medium is started. A constant rate, for example 1ml/h of fresh medium renders excellent results for many weeks with no need to subculture the cells.

Assembly of a working line for per(i)fusion culture

Screw cap innovative (Nr. 0004): At the top of the screw cap two drillings are present. The bigger one is for the filter, the smaller for the silicone tube. Please cut the end of the silicone tube so thin, that it fits through the drilling and cut the end.

Nr. 0004

IMPORTANT HINTS:

Important hints for sterilization: Using an autoclave the optimum is 105°C and 0.3 bar for 20 minutes. The absolute maximum is 135°C and 2 bar for 10 minutes. Most important, to avoid damage do not close the clamps of the culture container during the sterilization procedure! The national safty and biosafty instructions have to be taken into consideration.

Important hints for rinsing: The procedure for rinsing may varying depending on the cultured cells or tissues. After run incubate the used tools in destilled water. If necessary use then an ultrasound rinsing bath containing destilled water and rinse finally with warm water and a soft brush. Clean again with destilled water. If absolutely necessary a mild soap detergent may be used. Do not use abrasive cleaners or solutions such as alcohols, acetone or thinners etc.

Important: The national safety and biosafety instructions have to be taken into consideration!


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