SOP- Karnovsky Fixative
Last Updated: 9/25/06
Author: Kelli Church
References:
Electron Microscopy Sciences (www.emsdiasum.com)
Technical Data Sheet: Karnovsky’s Fixative
CAT #15720
Required Equipment:
Hood
Lab Coat
Gloves
Goggles
Four (4) Glass Pipettes
One (1) 9mm Pipette
Automatic Pipette Dispenser
One (1) 100 mL Beaker with Lid
Karnovsky’s Fixative Kit (CAT #15720)
Distilled Water, 32mL
Safety Information:
All procedures must be performed under Fume Hood
Keep track of all glass and disposed in glass waste container
Surfaces must be wiped down with 70% ethanol after process is completed
Store Karnovsky’s solution in refrigerator.
Procedure:
1) Place equipment underneath hood.
2) Carefully break glass bottles, by placing pressure on the small bulb at the top of the bottle with
your thumb, holding bottle steady index finger and other hand.
3) Break Glass away from face.
4) Use 9 mm pipette to remove solutions from glass bottles.
5) Mix the following in 100 mL Beaker:
16% Paraformaldehyde Solution: 20 mL
50% Glutaraldehyde EM Grade:
8 mL
0.2M Sodium Phosphate Buffer: 25 mL
Distilled Water
25 mL
6) Place Cap on solution.
7) Place solution into refrigerator with label (including CAT#, name, date) or Add Solution to larger
quantity already prepared
8) Dispose of glass waste in appropriate container.
9) Wipe down surfaces with 70% Ethanol