Professional Documents
Culture Documents
INTRODUCTION-
All the glass wares which are used for tissue culture should be clean and reserved
for that purpose alone. When glass wares are used either for storage or for
culture. The problem of contaminants leaking out into media or regents remain
and absolute cleanliness is therefore essential.
Material
Method-
2. clean the dust of new glass wares with the aid of a piece of cloth.
3.till the bottles, beakers with dichromate solution (potassium dichromate and
sulphuric acid in the mixture of 63gm of potassium dichromate and 35ml distilled
water cool it then add conc. sulphuric acid to mater in 1 liter.
4.Also immersed the other glass ware like pipette, burette, test-tube, beaker, etc.
in the dichromate solution an kept it immersed over night
5.remove the dichromate solution on the next day(this solution can be reused still
it changes it’s color
6.rinise the glass ware with clean water for several time and keep these glass
ware immersed in clean water over night and then follow the further steps for
further cleaning and washing
3.sort out the glass ware boil in water containing detergent for 10 min
4. allow the pan to cool till the water is Luke warm and then clean different atoms
as follows:
b)pipette-rinse for half and hour in the pipette rinse or pipette washer
c)needles check the point and sharpen it if necessary clean the needle with style
d)syringe is – scrub well with brush the pistol and barrel working smoothly
5.rinse the glassware in hot water or boil it in clean tap /distilled water
2.allow to cool and follow the steps as described for uninfected glass ware
2.remove the plate after ½ and hour and immerse in 3+hypoclorate solution For
at least 2 hours if necessary over night also.
9.after drying sterilize under uv light for 10 min at a distance for 12 inch of the
source.
Filter assembler:
2.clean the intermediate part of filter assemblies by scrubbing with brush and
water.
3.rise it with clean water for 10 times or with distilled water for 5 times.
6.assemble the filter paper and make it ready packing & sterilize.
Experiment no -3
Packing of glassware
4.if there is any deposit, patch or marks on any container send it back for on
second wash.
6.the plastic tips are loaded into tip holders and holders are wrapped in paper .
7.plug the upper end of the pipettes tightly with cotton wool(non-absorbent )
8.the conical flasks ,measuring cylinder ,tubes etc should be plugged with cotton
wool& envelope with cotton gauge.
9.cover the plugs of individual tubes , flasks etc with aluminum foil
12.sort out the pipettes & pack them in their respective conc.
13.bottem of the can should have cotton beds which should be changed at each
packing
14.use aluminum foil for covering sharp points or they may tear the brown paper
Sterilization of glassware’s
Types of sterilization
2.do not overload, leave sufficient gap between material for free movement of
hot air
3.sterilise at 160c for 1½an hour. precaution should be taken that the
temperature should not exceed 160c
4.check the steri . indicator tape color to ascertain that each container is
sterilized
5.first of all attain the dry air temperature above 155c for more than 30min
6.resterilise the material if there is any doubt but fix fresh steri. Tape to the
material.
1.solutions
2.rubberware
3.stopperes
4.washeres
5.screw caps, syringes
6.filtration assembly
1.all solutions except heat labile solution can be steri. By steam under pressure
4.rubber stopper, corn wall syringes filters otter instrument with rubber fittings
should be sterilized by autoclaving at 15 or10lbs at 120c or115c
5.place the material in autoclave care should be taken to losen the screw and
mark the mesious level
10.check the temp record the time & sterilize it for 15 min as per requirement
13.check the color of stir. Indicator tap & meniscus the sol.
Sterilization by UV light-
Principle:-
Medium is the most important in cell culture ideally the medium is an accurately
defined mixture of chemical substances .animal cell require the chemically
defined media to be supported with some mutual product e.g. serum. Tissue
extracts etc different all types different media.
The media required for animal culture in vitro must be prepared to contains
Certain factor necessary for survival of cell, cell growth and multiplication. These
are carbohydrate source, in organic irons ,gases, vitamins, hydrogen conc.etc
All these factors should be in aseptic condition through the cell culture media.
Media contain gin heal sensitive component are filter separately and then added
prior peruse this filtration must be under positive pressure of an inner gas like
nitrogen, it accerlate the filtration process in the
Requirement:-
The standard powder media ,distilled wear ,sodium bicarbonate co2 source
phenol sterinement filter assembly attach with succession assembly or pressure
assembly, sterile bottle, laminar flow cabinet etc.
Procedure:-