PMID: 4293006Sep 1, 1967Paper

Ultrathin frozen sections. II. Demonstration of enzymic activity

The Journal of Cell Biology
E H LeducJ P Tranzer

Abstract

Endogenous enzyme activity can be readily and routinely demonstrated in ultrathin, frozen sections for electron microscopy. The procedure employed to obtain the best structural preservation as well as enzyme activity in thin sections involved fixation in glutaraldehyde, embedding in thiolated gelatin or pure gelatin, partial dehydration in glycerol, and sectioning in a cryostat at -35 degrees C with a slightly modified Porter-Blum microtome on which the tissue is maintained at -70 degrees C and the knife at -23 degrees C. Kidney cortex was used as test tissue, but a few other organs were occasionally used. Thin sections were floated on the surface of several incubation media routinely employed for enzyme cytochemistry. Positive, specific reactions were obtained for alkaline phosphatase in kidney brush border, for adenosine triphosphatase in brush border and in basal membranes of distal tubules, for acid phosphatase and esterase in lysosomes, and for NADH diaphorase in mitochondria. Mitochondrial ATPase was sporadically evident only in the distal tubule of the kidney. Localizations of enzyme activity reported by other technical approaches were confirmed and in some cases somewhat improved.

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Citations

Jan 1, 1967·Histochemie. Histochemistry. Histochimie·E Pihl
May 1, 1976·The Histochemical Journal·O V Deimling, A Böcking
Jul 1, 1980·The Histochemical Journal·K T Tokuyasu
May 1, 1976·Journal of Ultrastructure Research·K T Tokuyasu
Sep 1, 1967·The Journal of Cell Biology·W Bernhard, E H Leduc
Jun 1, 1971·The Journal of Cell Biology·W Bernhard, A Viron
Jun 1, 1972·The Journal of Cell Biology·R BernierR Simard
Oct 1, 1975·The Journal of Cell Biology·E Puvion, W Bernhard
Mar 1, 1971·American Journal of Ophthalmology·K Lapis, M Radnót
Jan 1, 1978·Journal of Microscopy·W GeymayerY Hödl
Mar 7, 2009·Annals of Anatomy = Anatomischer Anzeiger : Official Organ of the Anatomische Gesellschaft·Wiebke Möbius
Jul 1, 1972·Journal of Ultrastructure Research·H Bauer, E Sigarlakie
Jan 13, 2006·Journal of Neuroscience Methods·Takumi AkagiKoujiro Tohyama
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Jan 1, 1969·Histochemie. Histochemistry. Histochimie·R Ahrens, N Weissenfels
Jan 1, 1972·Histochemie. Histochemistry. Histochimie·H Madreiter, O von Deimling

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