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This article gives a brief history of the evolution of the modern blood stain. Current theories of molecular mechanisms involved in the staining process are also reviewed.
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Explore the source record for details and available documents.
Explore the source record for details and available documents.
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Corneal histopathology is essential to understanding the mechanisms of corneal disease and wound healing. We have developed a light microscopic trichrome stain for 1-micron, aldehyde-fixed, osmicated, epon/resin-embedded corneal sections. The stain consists of azure II, methylene blue, and basic fuchsin. This combination of stains provides a fast and effective method to clearly differentiate corneal cells from their surrounding extracellular matrix, and chromatically separates various ECM materials within the cornea. Correlative examination by light and transmission electron microscopy (TEM) provides multilevel understanding of corneal morphology and ultrastructure. Tissues examined using this staining procedure may be subsequently examined by TEM for ultrastructural morphology. This stain has proved beneficial in our research of both cellular and extracellular constituents of the cornea.
Mast cells and histamine concentrations have been studied in uteri removed by hysterectomy from women in their post-menopausal years. Mast cell numbers were expressed as mean numbers/mm2 following fixation in 10% formalin and staining with Azure B. The majority of mast cells, in both the endometrium and myometrium, were very densely stained. Mast cells in the myometrium showed a significant negative correlation with years post-menopausal (rs = -0.52, P less than 0.05). Extracted histamine from uterine tissue was condensed with o-phthaldialdehyde to form a fluorophore and its fluorescence was measured at 450 microns using a spectrofluorometer. No significant correlation was found between histamine concentrations in the uterine wall and years post-menopausal.
A rapid sensitive method for the determination of Plasmodium falciparum in in vitro culture is presented. The technique employs a fluorescent flow cytometer equipped with a 15-mwatt argon laser that emits light at 488 nm and a membrane-permeable fluorochrome thiazole orange (TO) that stains RNA. Parasitized red cells are stained by suspending them in 1 ml of phosphate-buffered saline (PBS) containing 10(-5) M of TO and incubating this mixture for 15 min in the dark at room temperature. The stained cells may be analyzed fresh or after fixation with 1% paraformaldehyde/PBS or 0.25% glutaraldehyde/PBS. Alternatively the cells may be fixed first and then stained. There is excellent correspondence between the number of fluorescent-labeled parasitized red cells and Giemsa-stained cells.
Three staining techniques (Giemsa, Q-banding and R-banding) are used consecutively to localize the breakage points in chromosomes of human lymphocytes, irradiated during G2-phase with gamma-rays, at doses ranging from 50 to 200 rad. The large majority, about 85% of the breaks, occurs at the interbands, between R- and Q-bands. The discrepancy of this result, with regard to previously reported ones, is attributed to the strong bias of analysis when only one staining technique is used.
We performed staining experiments on cells using natural dyes and different mordants using techniques that are used for wool and silk dyeing. The natural dye sources were madder root, daisy, corn cockle and yellow weed. Ferrous sulfate, copper sulfate, potassium tartrate, urea, potassium aluminum sulfate and potassium dichromate were used as mordants. Distilled water, distilled water plus ethanol, heptane, and distilled water plus methanol were used as solvents. All dye-mordant-solvent combinations were studied at pH 2.4, 3.2 and 4.2. The generic staining procedure was to boil 5-10 onion roots or stimulated human lymphocyte (SHL) preparations in a dye bath on a hot plate. Cells were examined at every half hour. For multicolor staining, madder-dyed lymphocytes were decolorized, then stained with Giemsa. The AgNOR technique was performed following the decolorization of Giemsa stained lymphocytes. Good results were obtained for both onion root cells and lymphocytes that were boiled for 3 h in a dye bath that included 4 g madder root, 4 g ferrous sulfate as mordant in 50 ml of 1:1 (v/v) methanol:distilled water. The pH was adjusted to 4.2 with 6 ml acetic acid. We conclude that madder root has potential as an alternative dye for staining biological materials.
Loeffler's methylene blue, commonly used as a counter-stain for acid fast bacilli, was used to detect Helicobacter pylori in paraffin sections and touch smears of gastric mucosal biopsies from 15 patients with duodenal ulcer and 35 with non-ulcer dyspepsia, and the results were compared with the modified Giemsa stain. The time taken to stain smears by Loeffler's methylene blue was approximately 10 min and the results correlated well with those stained by the modified Giemsa stain. However, the Loeffler's methylene blue method was found to be simpler, quicker and cheaper than the modified Giemsa stain.
The sex chromosomes of the opossum, Didelphys virginiana, are the only elements that exhibit C-banding. In contrast, the sex chromosomes as well as the autosomes bear specific G-Bands. However, unlike other mammalian species different types of G-banding are observed if the chromosomes are pretreated with trypsin and SSC solution The SSC-pretreated chromosomes show discrete bands only when stained with Giemsa at certain pH values. An asynchronous pattern of terminal DNA replication is observed among the three C-banding regions of the X-chromosome. The inter- and intrapositive G-banding areas of the chromosomes are not always late in DNA replication in comparison to those negatively stained G-banding areas.
Additive reagents have been investigated to improve the stability of methanolic Wright's stain. The addition of ammonium halides, monoalkylamine hydrochlorides, dialkylamine hydrochlorides or trialkylamine hydrochlorides to methanolic Wright's stain was found to enhance the stability of stain components in methanol. No change in performance is observed with these additives present. Random precipitation in the stain solution was still observed with the addition of ammonium halides and monoalkylamine hydrochlorides. No precipitation was found in stain solutions containing hydrochlorides of most dialkylamines and trialkylamines. Of the compounds evaluated, 0.6% diethylamine hydrochloride added to methanolic stain solutions produced the most desirable overall results. Mechanisms of stabilization and precipitation in these stain solutions are proposed. Essentially, separation of the thiazine-eosinate ion pair through interaction with an appropriate additive increases stain stability. The solubilities of thiazine-eosinate or additive cation-eosinate ion pairs in methanol determine the formation of precipitate in such stain solutions.
Six different staining techniques were evaluated for their suitability to stain nuclei of Colletotrichum gloeosporioides f. sp. malvae (C.g.m.) spores. Of the three fluorescent stains, DAPI (4',6-diamidino-2-phenylindole) and bisbenzimide (Hoechst 33258) stained spore nuclei well; mithramycin did not. To achieve consistent results with the bisbenzimide staining protocol, the spores had to be fixed prior to staining and the stain had to be supplemented with Triton X-100. Both safranin O and Giemsa were suitable nonfluorescent staining techniques; lomofungin was not. Safranin O staining was simple and rapid. However, reproducibility was better if the spore suspension and KOH droplets were rapidly mixed prior to adding the stain. There was no significant difference in the percentages of uninucleate and binucleate spores observed in spore preparations stained with DAPI, bisbenzimide, safranin O or Giemsa. Bisbenzimide and safranin O were found to be simple, rapid and reliable fluorescent and nonfluorescent techniques, respectively, for staining nuclei of C.g.m. spores.
Cultures of bone marrow obtained from patients with various forms of leukemia have been stained by the Giemsa-11 technique. The results suggest that this method can be utilized for the constant monitoring of numerical and structural anomalies in chromosome 9. Furthermore, by varying the pH, this technique can be applied not only to freshly prepared slides but also to stained preparations stored for many months.