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Table 5 Postfixation procedures reported by the included studies

From: Perfusion fixation in brain banking: a systematic review

Study Pre-processing Fixative Buffer Temp Length of postfixation
Adickes 1996 [2] NR 10% buffered formalin Phosphate NR 1 day (if postfixed)a
Adickes 1997 [1] Cut into 1–1.5 cm-thick sections 10% buffered formalin Phosphate NR 5–6 h
von Keyserlingk 1984 [93] Brain left in skull for 2 h, then removed and dissected 1% osmium tetroxide 0.1 M sodium cacodylate NR 2 h
Istomin 1994 [43] NR 10–12% formalin Neutral-buffered NR NR
Beach 1987 [7] NR 4% paraformaldehyde 0.1 M phosphate buffer (pH 7.4) 4 °C NR
Benet 2014 [9] NR 1:10 dilution of 10% formaldehyde NR 5 °C > = 2 days
Benet 2014 [9] NR 1:10 dilution of 10% custom solution (ethanol 62.4%, glycerol 17%, phenol 10.2%, formaldehyde 2.3%, and water 8.1%) NR 5 °C > = 2 days
Böhm 1983 [12] Cut into 1 cm-thick coronal sections Paraformaldehyde or formalin 0.1 M phosphate buffer NR NR
Coveñas 2003 [20] NR 4% paraformaldehyde 0.15 M PBS (pH 7.2) 4 °C 30 days
de Oliveira 2012 [23] NR 20% formalin NR NR 3 weeks
Donckaster 1963 [24] Brain removed Cajal fixative: formalin and ammonium bromide NR NR 4 days
Grinberg 2008 [34] NR Same fixative as was used for fixation NR NR NR
Huang 1993 [40] Dissection of brainstem NR NR NR <= 24 h
Insausti 1995 [42] Dissected into slabs approximately 1 cm thick 4% paraformaldehyde NR NR 48–72 h
Kalimo 1974 [45] (electron microscopy) Brain left in the skull for 1 to 2 h after perfusion fixation, then removed, then samples dissected for EM 1.0% paraformaldehyde, 2.0% glutaraldehyde 0.1 M cacodylate (pH 7.4) NR Overnight
Kalimo 1974 [45] (histology) Same as above 10% formaldehyde NR NR 10 days
Latini 2015 [53] Brain extracted from the skull 48 h after perfusion 10% formalin NR NR 24 h
Lin 2000 [57] NR 4% paraformaldehyde 0.1 M phosphate buffer (pH 7.4) 4 °C Overnight
Lyck 2008 [58] Brain removed from skull 4% paraformaldehyde 0.15 M Sørensens phosphate buffer (pH 7.4) 4 °C 2 weeks
Masawa 1993 [59] NR 4% formalin 0.1 M phosphate buffer NR > = 3 days
Masawa 1993 [59] (electron microscopy) From postfixed tissue, tissue blocks were cut and buffer washed 1% osmium tetroxide solution NR 4 °C 90 min
McGeer 1988 [62] NR 4% paraformaldehyde NR NR 2–3 days or until the pink color of unfixed erythrocytes was gone
McKenzie 1994 [64] Waited 1 h after perfusion fixation, then the skull was opened, and the brain was removed Formalin Neutral-buffered 4 °C NR
Pakkenberg 1966 [70] Brain removed from skull Alcohol 80% 9 parts, formalin 4% 1 part NR NR 3 weeks
Sharma 2006 [79] Brain suspended in a bucket 20% formalin Neutral-buffered NR 1–4 days
Shinkai 1976 [81] Cut into 2 mm-thick tissue blocks 2.5% glutaraldehyde containing 0.2 M sucrose NR NR 4–8 h
Sutoo 1994 [85] Brain halved sagittally and sliced into 10 mm coronal blocks 4% paraformaldehyde PBS 4 °C 2 days
Suzuki 1979 [86] Dissected bifurcations of the first temporal branches of the middle cerebral arteries 2.5% glutaraldehyde NR NR 4 h
Tanaka 1975 [87] (electron microscopy) Samples taken from various regions of the brain 1.0% osmium tetroxide NR NR NR
Tanaka 1975 [87] (histology) Rest of the brain 8.0% formaldehyde NR NR NR
Torack 1990 [90] Hippocampus and entorhinal cortex was isolated and sectioned into 0.5 cm thick slices 4% paraformaldehyde +/− 1% Bouin’s solution (picric acid, acetic acid, and formaldehyde) NR NR 48 h
Turkoglu 2014 [92] Brain removed from skull 10% formaldehyde NR NR 2 weeks
Waldvogel 2006 [96] NR 15% formalin 0.1 M phosphate buffer (pH 7.4) NR 6–12 h
Welikovitch 2018 [99] Dissected out the medial temporal lobe 4% paraformaldehyde and 0.2% picric acid 0.1 M phosphate buffer NR Overnight
  1. a: Note that in Adickes et al. (1996), the brain is either cut immediately or postfixed in formalin for one day. NR: Not reported, PBS: Phosphate-buffered saline