Acrolein Fixatives (from: MILLONIG G.(1976): Laboratory Manual of Biological Electron Microscopy (Publ.: Mario Saviolo – Editore – Vercelli – C.P. 182, Italia) (Millonig G. Ed., p -28-) CAVE: TOXIC – prepare under hood and use PSA Fixation time (@RT): 30-120 min
Method 1 [50]: 10 % Acrolein (in 0.1 M phosphate or cacodylate buffer) Buffer 0.2 M pH 7.6 A. bidest./H2O Acrolein
25.0 ml 20.0 ml 5.0 ml
Method 2 [14]: 1 % Acrolein + 2.5% Glutaraldehyde (in 0.1 M buffer ) Buffer 0.2 M pH 7.6 A. bidest./H2O Glutaraldehyde 25% Acrolein
25.0 ml 20.0 ml 5.0 ml 0.5 ml
Method 3 [19]: 2% PFA + 3 % Glutaraldehyde + 1 % Acrolein + 2.5% DMSO (in 0.1 M buffer) 2% PFA in (0.1 M) Cacodylate buffer Glutaraldehyde 25% Acrolein Dimethylsulfoxide (DMSO) CaCl2 (= final Concentration= 0.001 M) Procedure: Primary Fixation: Rinse: buffer 0.1M + 0.1M sucrose (= 3.4 %) Secondary Fixation: 2% OsO4 in 0.2M Sucrose (= 7.0 %) Rinse: buffer 0.1M Dehydration - Embedding
44.0 ml 5.0 ml 0.5 ml 1.25 ml 0.008 g up to 16 hrs 6 x 15 min 4h
Method 4 [15]: 1.5 % PFA + 3 % Glutaraldehyde + 1.5 % Acrolein (in ~0.08 M buffer) 6% PFA in A. dest. 10% Glutaraldehyde 10% Acrolein 0.2 M Phosphate or Cacodylate buffer A. dest. (H2O) References ./.
3.0 6.0 3.0 5.0 1.0
ml ml ml ml ml
References: [14]: Glauert AM (1975): Fixation, Dehydration and Embedding of Biological Specimens. North Holland/American Elsevier
[15]: Hayat MA (1970): Principles and Techniques of Electron Microscopy. Vol.1: Biological Applications. Van Nostrand Reinhold Publ. Co
[19]: Kalt MR, Tandler B (1971): A Study of Fixation of Early Amphibian Embryos for Electron Microscopy. J. Ultrastruct. Res. 36 (5, Sep): 633-645 [personal Note: acc. to PubMed: cf. http://www.ncbi.nlm.nih.gov/pubmed/4939156?dopt=Abstract ]
[50]: Sabatini DD, Miller F, Barrnet RJ (1964): Aldehyde Fixation for Morphological and Enzyme Histochemical Studies with the Electron Microscope. J. Histochem. Cytochem. 12, p.57 – 71 Compilation from the original booklet (12-10-01): Dr. Wolfgang MUSS, EM-Lab, Pathology SALK-LKH (Gen. Hospital) SALZBURG, Austria (
[email protected])
