- SUBJECT: Recipe for buffered formalin
- DATE: 2/97
-
- Hi labbers- Could someone tell me how to make neutral buffered formalin?
- Thanks. Mary
-
- Mary H. Gray D.V.M.
- Department of Biological Sciences
- Lilly Hall of Life Science
- Purdue University
- West Lafayette, IN 47907
- (765)-494-8185
- FAX (765)-494-0876
-
-
- Hope this is useful
-
- Neutral Buffered Formalin (pH 7.0)
-
- Formalin, full strenghth (37-40% formaldehyde) 100.0 ml
- Sodium phosphate dibasic (anhydrous) 6.5 gr
- Sodium phosphate monobasic 4.0 gr
- Distilled water 900 ml
-
- from "A Manual of Histotechniques" 3rd ed. Ann Preece, 1972.
An old text but
- I doubt if the formula has changed any.
-
- Al
-
-
- % Al Williams AAWilliams@Manchester.edu
- % Biology Department Voice 219-982-5308
- % Manchester College FAX 219-982-5043
- % N. Manchester, IN 46962
-
-
-
- Mary,
-
- I use the following recipe to prepare formaldehyde from paraformaldehyde
for
- fixing mammalian cells for immunofluorescence studies:
-
- 1. Heat 15 mls of PBS (phosphate buffered saline, pH 7.2)(see below
for PBS
- recipe) to about 55 degrees C.
-
- 2. With stirring, add 0.75 grams paraformaldehyde to the PBS.
-
- 3. Add, dropwise, 0.1 N NaOH (the combination of heat and NaOH breaks
the
- paraformadehyde polymer into formaldehyde molecules) just until the
solution
- becomes completely clear.
-
- 4. Let cool and adjust the pH to desired value.
-
- 5. Add an additional 10 mls of PBS.
-
- PBS recipe (I need to use a Mg- and Ca-free variety):
- -----------------------------------------------------
- KCl 0.2 grams
- KH2PO4 0.2
- NaCl 8.0
- Na2HPO4*7H2O) 2.16
-
- Final volume to 1L with distilled water.
- -----------------------------------------------------
-
- Hope this helps. Good luck.
-
- George
-
- George Edick
- RPI - Dept. Biology
- Troy, NY 12180
- edickg@rpi.edu
-
-
-
- The recipe for buffered formalin from Humason is:
-
- 10% formalin (stock is 37%) 1000 ml
- NaH2PO4.H2O 4 g
- Na2HPO4 6.5g
-
- May remain in fixative indefinitely. Action is progressive
-
- From: greggk@winthrop.edu
-