The Localization of Acid Phosphatase in Rat Liver Cells as Revealed by Combined Cytochemical Staining and Electron Microscopy

Abstract

Discrete localization of stain in pericanalicular granules was found in 10 µ frozen sections of formol-phosphate-sucrose-fixed liver stained by the Gomori acid phosphatase technique and examined in the light microscope. The staining patterns, before and after treatment with Triton X-100 and lecithinase, were identical with those previously reported for formol-calcium-fixed material treated in the same way, and it can be assumed that the stained granules are identical with "lysosomes." Examination in the light microscope of the staining patterns and lead penetration in fixed blocks and slices of various dimensions showed nuclear staining and other artefacts to be present, produced by the different rates of penetration of the various components of the staining medium into the tissue. A uniform pericanalicular staining pattern could be obtained, however, with slices not more than 50 µ thick, into which the staining medium could penetrate rapidly from both faces. The staining pattern produced in 50 µ slices was the same both at pH 5.0 and pH 6.2, and was not altered by subsequent embedding of the stained material in butyl methacrylate. Electron microscopy showed the fine structure of fixed 50 µ frozen slices to be well preserved, but it deteriorated badly when they were incubated in the normal Gomori medium at pH 5.0 before postfixing in osmium tetroxide. After incubation in the Gomori medium at pH 6.2, the detailed morphology was substantially maintained. In both cases lead phosphate, the reaction product, was found in the pericanalicular regions of the cell, but only in the vacuolated dense bodies and never in the microbodies. Not every vacuolated dense body contained lead, and stained and unstained bodies were sometimes seen adjacent to each other. This heterogeneous distribution of stain within a morphologically homogeneous group of particles is consistent with de Duve's suggestion (9) that there is a heterogeneous distribution of enzymes within the lysosome population. It is concluded from these investigations that the vacuolated dense bodies seen in the electron microscope are the morphological counterparts of the "lysosomes" defined biochemically by de Duve.

Changes in Fine Structure and Acid Phosphatase Localization in Rat Thyroid Cells Following Thyrotropin Administration

“…For fine structural localization of acid phosphatase, tissues were fixed at 4°C for 18 hours in either 10 per cent formalin containing 2 per cent calcium acetate, or sucrose phosphate formalin (Holt and Hicks, 1961), or for 2 hours in the glutaraldehyde solution prior to processing according to methods described by Holt andHicks (1961) andMiller (1962). After fixation, tissues were rinsed 3 times for 5 minutes each in 7.5 per cent sucrose at 4°C, frozen quickly in this solution on a microtome hammer with dry ice, and sectioned at 40 tz in the cryostat.…”

Localisation ultra-structurale de l’activité oxydasique chez Toxoplasma gondii

“…Il a d'ailleurs été constaté depuis longtemps (Barka et Anderson, 1962 ;Holt et Hicks, 1961) que la réaction des phosphatases acides par la méthode de Gomori devait être interprêtée avec prudence étant donnée la fréquence des fausses réactions positives (précipités ou simple ombrage au plomb des structures cellulaires). Par contre une très faible quantité d'enzymes peut être présente dans le parasite sans être révélée par cette réaction relativement peu sensible aux faibles concentrations de phosphatase acide (Barka et Anderson, 1962 ;Holt et Hicks, 1961).…”

“…Par contre une très faible quantité d'enzymes peut être présente dans le parasite sans être révélée par cette réaction relativement peu sensible aux faibles concentrations de phosphatase acide (Barka et Anderson, 1962 ;Holt et Hicks, 1961).…”

Immunocytochemical localization of acid phosphatase in rat liver.

“…In addition, we found ACPase in endosomelike vesicles, which are present in the cytoplasm around the bile canaliculus and just below the plasma membrane of the sinusoidal surface. A positive reaction for ACPase in the endosomelike vesicles has not been clearly shown by the enzyme cytochemistry (4,13,20). The present study showed that ACPase is associated with the plasma membrane and with the lysosomal membrane, as reported by Parenti and coworkers (19).…”

“…After the pioneering work of Sheldon and coworkers (24), the "lead capture" method of Gomori (10) was adapted to electron microscopy, and the ultrastructural localization of ACPase in various tissues and cells under normal and pathological conditions has been since described by many investigators (2,4,7,13,18). ACPase seems to be one of the most extensively studied enzymes that are cytochemically demonstrable.…”