1. Measure 100ml Phosphate Buffered Saline (PBS) into a measuring cylinder. Pour into the conical flask containing 4g of paraformaldehyde. Cover with parafilm and transfer to the fume hood: thoroughly shake - take care not to splash paraformaldehyde about - it is a rapid fixer and is TOXIC.
2. Place flask on top of the hotplate/stirrer inside the fume cupboard and set the heat control to 7 with moderate stirring. Allow the solution to warm up-it will turn from being cloudy to clear when ready. Inspect regularly to avoid over- heating and consequent spilling.
3. When the paraformaldehyde has dissolved, switch off the heat but leave to stir: do not handle for safety reasons. Allow to cool.
4. When cooled, transfer the fixative to a 4C refrigerator. Label appropriately and date.
0.4% PARAFORMALDEHYDE/PBS (FOR POST-DIGESTION FIXATION)
This fixative solution should be made up fresh.
Dissolve 0.4g paraformaldehyde/100ml DEPC-treated PBS in a conical flask and seal with Sealon film. On a heated stirrer (set to 6-7), inside the fume hood, mix until the milky solution turns colourless. Any cloudiness may be removed by adding a drop of 5M sodium hydroxide. Allow to cool before handling.
Paraformaldehyde is a rapid fixative and will cause harm if it comes into contact with the body.
WEAR GLOVES AND GOGGLES
PLEASE REFER TO THE APPROPRIATE SECTION IN THE CONTROL OF SUBSTANCES HAZARDOUS TO HEALTH (COSHH) REGULATIONS WHEN PREPARING/HANDLING THESE REAGENTS
NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.
© Copyright 1997 University of Nottingham Medical School Division of Histopathology. This page was last built on Mon, May 5, 1997 with Frontier. Thanks for looking in. Comments to James.Lowe@nottingham.ac.uk