Modified Staining Technique Research Articles

Cytochemical study of macrophage lysosomal inorganic trimetaphosphatase and acid phosphatase

Cytochemical investigations have associated acid inorganic trimetaphosphatase (TMPase) activity with the lysosomes of certain cell types. We have used the modified staining technique of Berg to show that this enzyme activity is present in normal mononuclear phagocytes and macrophage cell lines. We have found this enzyme activity to be present in murine RAW264 macrophages, in human U937 macrophages, in normal human blood monocytes, and in guinea pig peritoneal macrophages. All of the RAW264 and U937 macrophages showed intense TMPase activity. Many of the human monocytes and most of the guinea pig macrophages were labeled by this method. The reaction product was associated with the lysosomes of these cell types. The lysosomal staining pattern was similar to that of acid phosphatase. Differences with regard to Golgi staining were noted. This indicates that TMPase is a lysosomal enzyme of mammalian macrophages. The distinction between TMPase and acid phosphatase activity has been demonstrated by measuring the pH optimum of each enzyme. Using substrates identical to those of the ultrastructural cytochemistry, we show that the pH optimum of TMPase is 4.0 and that of acid phosphatase is 5.0. The enzymatic activities are therefore ultrastructurally and biochemically distinct. Following phagocytosis of latex, yeast ( Saccharomyces cerevisiae), or Corynebacterium parvum), TMPase has been found to be associated with phagosomes. This enzyme may take part in the degradation of phagocytosed materials, particularly microorganisms which contain inorganic polyphosphates and metaphosphates.

Haematology and blood chemistry of the red kangaroo Megaleia rufa in captivity.

Haematology and biochemistry was performed to establish base line values on not less than 30 samples of blood and serum from 10 red kangaroos Megaleia rufa held in captivity. The kangaroos were apparently healthy. Ranges for results were: Haemoglobin 16-18 gm/100 ml, PCV 46-51%, erythrocytes 4.9 - 5.7 times 10(6)/mm3. MCV 86-98 mum3, MCH 30-34 pg, MCHC 34-36%, leucocytes 3.8 - 7.1 times 10(3)/mm3, neutrophils 2.0 - 4.7 times 10(3)/mm3, lymphocytes 1.2 - 2.4 times 10(3)/mm3, monocytes 0.1 - 0.3 times 10(3)/mm3, eosinophils 0.06 - 0.013 times 10(3)/mm3, SGOT 36-43 u/l, SGPT 30-41 u/l, LDH 30-85 u/l, protein 6.0-6.8 g/100 ml, albumin 2.8 - 3.4 g/100 ml, alpha globulin 0.3 - 0.7 g/100 ml, beta globulin 0.7 - 0.9 g/100 ml, gamma globulin 1.7 - 2.3 g/100 ml, albumin/globulin 0.8 - 1.1, Na+ 146-152 mEq/l, K+ 4.6 - 5.6 mEq/l. Rectal temperature was 36.8 +/- 0.3. High haemoglobin may be characteristic of wild animals and of animals which have a high level of muscular activity and large erythrocytes may be representative of marsupials. Haematology as a tool of ecological investigation and its role in future research is briefly discussed. A modified staining technique for performing differential blood cell counts is described.

A Modified Staining Technique for Carbonates in Thin Section

POTASSIUM ferricyanide and alizarin red S are frequently used as stains for distinguishing between carbonate minerals. Potassium ferricyanide in acid solution produces Turnbull's blue with ferrous iron and it used to be thought that dolomite could be distinguished from calcite on the basis that dolomite alone contained ferrous iron. Recently this has been shown to be incorrect1, and it is now known that any carbonate mineral containing small quantities of ferrous iron will stain with potassium ferricyanide providing the carbonate can be made to react with an acid medium. Recent work2,3 shows that alizarin red S in solution is probably the best method of distinguishing between the common carbonates.

Regeneration of gastric mucosa in rats

Since Bensley's ( 1898) classical description of the specialized cell types in the gastric mucosal epithelium much uncertainty has existed concerning their life history. Grant ('45), Stevens and Leblond ('53), Hunt ('57) and Messier ('60) observed that both surface and mucous neck cells arise from the proliferation of cells in the neck region of the fundic gland and are sloughed off, the mucous neck cells at their site of origin and the surface cells after moving to the surface of the mucosa. Hunt ( '57) and Grant ('45) concluded that other cell types in the glands are also lost. Harvey ( '07), Ferguson ('28), and Hunt ('58) implied that parietal and chief cells are replaced by transformation from mucous neck cells but Stevens and Leblond ('53) and Messier ('60) denied the loss and replacement of these cell types. In order to acquire more precise information concerning the origin of the various cell types in fundic glands, numerous investigators have studied their re-appearance in gastric mucosa following surgical or thermal injury to it (Harvey, '07; Ferguson, '28; Gunter, '50; Williams, '53; Hunt, '58; Finckh and Milton, '60). These investigations revealed that (a) the first epithelial cells growing back into an injured area contain mucus and, at least in the newly-formed glands, come to resemble mucous neck cells except for their shape and nuclear structure; and (b) the newlyformed chief and parietal cells seem to arise from these mucus-containing cells. However, conclusive evidence for direct transformation of mucous cells to chief and parietal cells is lacking as is also information concerning the origin of argentophilic cells. Until such evidence is available the sources of these cells cannot be considered proven. Much of the difficulty in visualizing these transformations has arisen from the lack of techniques adequate for the demonstration of transitional forms. The objective of this study was to secure cytological evidence concerning the cellular origin of the chief, parietal and argentophilic cells as they arise in regenerating fundic stomach, utilizing newly modified staining techniques.