Light microscopic studies on the distribution of glycogen in human liver biopsies with special reference to the limiting plate.
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Adult↗
Biomedical subjects
H Lyon
Publications and source records attributed to H Lyon.
Standardized methyl green-pyronin Y procedures using pure dyes.
Fully standardized Methyl Green-Pyronin methods are presented. Pure Pyronin Y and purified Methyl Green or Ethyl Green are used either simultaneously in one dye bath or are used as a sequence of Pyronin Y and Ethyl or Methyl Green. Both methods, as shown by enzymatic pretreatment, give a reliable and reproducible staining in DNA with Ethyl or Methyl Green and of RNA with Pyronin Y on Carnoy fixed material. On formaldehyde fixed material it was found advantageous to use the sequential method as chromatin was hereby stained green instead of blue as seen with the simultaneous method.
Animals↗
Quantitation of dye binding by cell monolayers in a microtiter system.
A semi-automated system has been developed for the quantitation of dye binding to cultured eukaryotic cells. It is based on staining precisely controlled numbers of cells seeded into microtiter trays. Cell-bound stain is then released using an appropriate solvent and quantitated in situ by measuring absorbance in a single beam ELISA reader with an interactive microcomputer link. In order to illustrate potential applications of this approach, the time course of dye-monolayer association and influence of cell number and stain concentration on staining has been examined for four dyes, Crystal Violet, Naphthol Yellow S, Ethyl Green and Pyronin Y. In addition, the effect of sequential and simultaneous staining was examined for Ethyl Green and Pyronin Y. The results provide evidence for the overall reliability of this approach as well as revealing several interesting features in the individual procedures examined. The combination of microtiter technology and computer link make the system particularly well suited to the efficient investigation of the permutations involved in optimizing conditions for a given staining procedure, as well as analysis of the thermodynamics of dye substrate interaction. Overall, the approach is viewed as an intermediate between artificial gel systems and microdensitometry.
Animals↗
An investigation of new commercial samples of methyl green and pyronin Y.
New commercial samples of Methyl Green (Gurr Certistain), Pyronine G (Gurr Certistain) and Pyronin Y (Polysciences) have been investigated using spectrophotometry, thin layer chromatography and nuclear magnetic resonance, in addition to standardized simultaneous and sequential staining methods using purified Ethyl Green and pure Pyronin Y as reference dyes. The Methyl Green was found to be Ethyl Green contaminated with Crystal Violet. It did not have any advantages compared with Ethyl Green supplied by American dye companies. The Pyronine G sample was Pyronin Y with a high dye content that gave good staining results when used with purified Ethyl Green. Pyronin Y from Polysciences was found to be essentially pure Pyronin Y.
Chromatography, Thin Layer↗
Induced fluorescence of serotonin in paraffin sections from carcinoid tumors.
The detection of argentaffin cells and carcinoid tumor cells would be greatly facilitated if it were possible to apply methods of induced fluorescence directly on paraffin sections. We have subjected 5 microgram thick sections from paraffin blocks containing carcinoids to different procedures for induced fluorescence. These include formaldehyde vapor at 80 degrees C for 1 hour, 5% paraformaldehyde solution at room temperature followed by heating to 80 degrees C for 15 min, and sucrose-phosphate buffer-glyoxylic acid solution either for 3 seconds at room temperature followed by heating to 80 degrees C for 15 min or 15 min at 60 degrees C in the solution. The different procedures were applied to sections which were non-deparaffinized, deparaffinized or deparaffinized and rehydrated to various degrees. With all procedures used we were able to induce histofluorescence of argentaffin cells and carcinoid tumor cells, and it was confirmed by microspectrofluorometry that the fluorophore originated from serotonin. Best results, when morphology was taken into consideration, were obtained with sections deparaffinized, transferred to 96% ethanol and then exposed to formaldehyde vapor.
Carcinoid Tumor↗