Journal of Histochemistry & Cytochemistry

Colvin, J R; Sowden, L C; Male, R S

doi: 10.1177/23.5.1133455pmid: 1133455

The relative iron, copper and mercury contents of individual, isolated erythrocytes from eight people were determined by analytical electron microscopy. The variation in iron content between erythrocytes of the same sample is more than six times, for copper content more than tem times and for mercury more than five times. Similar variations were observed for 1-day-old chick defintive erythrocytes and for 4-day-old chick embryo primitive erythrocytes. The range of variation does not depend greatly, it at all, on the age of the erythroyctes or the tissue of origin. There is little or no correlation between the variation of iron contnet and that of copper. The cause of the wide variation of metallic ion contnt among erythroyctes is not yet known.

Linde, A; Magnusson, B C

doi: 10.1177/23.5.165233pmid: 165233

The effects of the alkaline phosphatase inhibitors levamisole and R 8231 on p-nitro-phenylphosphatase, inorganic pyrophosphatase and adenosine triphosphatase (ATPase) activities in dentingenically active odontoblasts were studied. The p-nitrophenylphosphatase and inorganic pyrophosphatase activities were inhibited, while 40% of the ATP-splitting enzyme activity remained under the assay condition used. This finding, togeather with earlier studies, indicates that at least two different phosphatase are active at alkaline pH in hard tissue-forming cells; on nonspecific alkaline phosphatase and one specific ATPase. The ATPase activity is uninfluenced by ouabain and ruthenium red and is activated by Ca-2+ ions.

Mehard, C W; Volcani, B E

doi: 10.1177/23.5.1127221pmid: 1127221

Chemical, radiochemical and x-ray microanalysis assays were used to define parameters of silicon (Si) retention during preparation og biologic samples (rat liver, spleen, kidney, lung, diatoms and cell organelles) for x-ray microanalysis, Due to its longer half-life 68-Fe was used in some cases to trace SI. Leaching of Si from cells and organelles by the aqueous preparation media was overcome by use of the freeze-substitution process. Cells were treated with 30% glycerol hypertonic sucrose medium to reduce ice damage. Embedment in Spurr's low viscosity epoxy resin medium caused no apparent Si loss. A semiquantitative evaluation showed 0.5 x 10-8 to 0.3 x 10-17 g detectable Si in isolated rat liver mitochondria in thin sections, which is within the instrument's range of detection. This study indicateds that the presence of Si in the mitochondria is not the rsult of contamination.

Gulys, B J; Yuan, L C

doi: 10.1177/23.5.805170pmid: 805170

Microperoxisomes were identified in the franulosa lutein cells from the corpora lutea of rhesus monkeys (Macaca mulatta). These organelles were histochemically visualized in aldehyde-fixed tissues icubated in alkaline 3,3'-diaminobenzidine (DAB). The DAB staining of the microperoxisomes was abolished when the tissues were preincubated in specific inhibitors for catalse or when the H2O2 was omitted from the DAB medium. Microperoxisomes were differentiated from primary lysosmes by the Gomori acid phosphatase staining. Tortuous undulating agranular endoplasmic reticulum (ER) was usually closely associated with microperoxisomes. Those regions of the granular ER which were closely associated with microperoxosomes lacked ribsomes. Micropersoxisomes were often contiguous with lipid droplets, and in some instances the limiting membrane of the moroperosisomes appeared discontinous at the point of contiguity, and the DAB staining substance diffused onto the surface of the lipid droplet. In these instances, the adjacent area of the lipid droplet showed electron-lucent staining.

Dunlap, L A; Warters, R L; Leif, R C

doi: 10.1177/23.5.1127222pmid: 1127222

This paper describes the modification of Centrifugal Cytology for the preparation of permanent, fixed, stained dispensions for both light and scanning electron microscopy of cells which have been isolated on bovine serum albumin (BSA) boyant density gradients. The principal problem with BSA gradient fractions is that the albumin which is present even after dilution is precipitated by the glutaraldehyde fixative. This problem has been solved by the layering of an intermediate D2O solution under the BSA and subsequent removal of the BSA solution and the underlaying with D2O containing glutaraldehyde. A special layering machine facilitates and expedites these operations. This technique has also been applied to BSA-seperated guinea pig and chicken bone marrow cells, as well as Ehrlich ascites tumor cells, hen and human blood cells. The number of celll present in each area of the slide is maintained at a constant value by utlizing a table of dilution factors. This table was generated by a computer program which calculates the concentration of cells present in the rractions and divides it by the number of celll desired.

Leif, R C; Smith, S B; Dunlap, L A; Leif, S B

doi: 10.1177/23.5.1127223pmid: 1127223

Guinea pig bone marrow cells were separated by buoyant density utilizing linear gradients of bovine serum albumin (BSA). It has finally become possible to characterize the cells present in the density fractions in terms of classical morphology. The development of the Cell Type computer program which calculates the percentages of the individual types of cells present in the fractions and their buoyant density distributions and plots the data has greatly facilitated and improved the accuracy of these studies. Approximately 40 cell types were observed in guinea pig bone marrow. Cells with definitive morphologies such as erythrocytes, the neutrophilic series, the binucleate blast megakaryocyte precursor and cells in mitosis band as virtually single peaks. Cells which are parts of continua or can easily be wrongly classified are found in multiple peaks. The small lymphocytes which are known to be polydisperse are found as five peaks. Because of the very strong benzidine staining by the glutaraldehyde-fixed hemoglobin, some of the erythroblasts were wrongly staged, resulting in a multimodal distribution. The presence of macrocytes further complicated these distributions. The rule that the younger cells are always less dense than the mature cells was adhered to in those cases where the cells could be definitively characterized, such as the neutrophilic series and the blasts. These results indicate that morphology is a good first approximation of reality.

Swift, A J

doi: 10.1177/23.5.47874pmid: 47874