A Microwave Oven Method for the Combined Alcian Blue-Periodic Acid-Schiff Stain

Abstract

A rapid microwave modification of alcian-blue-periodic acid-Schiff stain is described. The method gives consistent and reliable staining of acid and neutral mucins within 15 minutes. (The J Histotechnol 12:295, 1989)     

A Warthin-Starry Method for Spirochetes and Bacteria Using a Microwave Oven

Abstract

A staining method for demonstrating spirochetes and bacteria, which stain poorly with the Gram stain, is described. The method is similar to the Warthin-Starry procedure except that the silver impregnation and reducing steps are done in a microwave oven and an acetic acid-glycine solution is used in place of citric acid. If spirochetes are to be stained, the sections are treated with uranyl nitrate, which is also heated in a microwave oven. The staining results are consistent and reliable. (The J Histotechnol 11: 149, 1988.)     

Volume 13, March 1990—December 1990

Abstract

A microwave Giemsa method for paraffin embedded tissue sections is reported. It is a modification of an earlier stain by Churukian and Listinsky, in which warm glycerol is used to dissolve the dyes and Triton X-100 is added to the final solution. The procedure can be completed within one hour and fifteen minutes and is consistent and reliable for demonstrating a variety of cells and microorganisms, like rickettsia and gram-negative bacteria. The stain can be easily prepared in the laboratory, does not require a microscope to control differentiation, and is as good, if not better, than commercially available Giemsa stains. Results are more uniform and consistent than those obtained with the widely used Wolbach method. (The J Histotechnol 18:319, 1995).     

A Toluidine Blue Method for Demonstrating Mast Cells

Abstract

A modified Giemsa azure-eosin method for staining polymethyl methacrylate (PMMA) plastic embedded bone sections is described. The method is similar to those used to stain glycol methacrylate sections but differs in timing and addition of air drying steps. The stain yields effective differentiation of marrow cells, growth plate, calcified cartilage, osteoid, bone matrix and bone cells in undecalcified sections of bone. The stain does not interfere with the resolution of fluorochrome labeled specimens. Hence, the same slide can beused to evaluate static and dynamic parameters of bone metabolism. (The J Histotechnol 14:85, 1991)     

The Denn-Golgi stain — A Modified Defano's Silver Impregnation Stain for Dendritic Spines and Neurofibrils

Abstract

A modification of Defano's method for Golgi apparatus is described with emphasis on reduced overall stain time and better stain definition. This method utilizes a variation of widely used silver nitrate impregnation solution modified for a specific research need. The addition of a counterstain provides improved contrast and definition of dendritic spines in the Golgi apparatus of brain tissue. This modified method provides consistent, reproducible results for large numbers of brain specimens, and as such is believed to be superior to the Defano method.     

Commercially available fluorescein-conjugated monoclonal antibody for determining the in vitro activity of antimicrobial agents againstChlamydia trachomatis

A simple, reproducible method for determining the antibiotic susceptibility of chlamydial isolates is described. Minimum inhibitory and lethal concentrations (MICs and MLCs) were determined for tetracycline and erythromycin titrated against a recent clinical isolate of Chlamydia trachomatis in McCoy cell cultures. A fluorescent antibody stain was found to be more sensitive than giemsa staining, generally giving two-fold higher values for both MICs and MLCs.     

Modified Movat's Pentachrome Stain

Abstract

We modified Movat's pentachrome stain for elastic fibers by substituting an iron-hematoxylin stain for resorcin fuchsin. The method yields consistent results, provides better elastic fiber staining, and reduces the time required from overnight to approximately two and one-half hours.     

A Tinctorially Distinctive Hematoxylin and Eosin Stain with Relative Simplicity and Consistency

Abstract

Paraffin embedded sections fixed in 10% formalin were stained in Shandon Instant Hematoxylin, differentiated in acid alcohol, blued in dilute sodium acetate, and counterstained with a concentrated eosin-multichrome stain. This simple procedure produced consistent results with deep blue nuclei and bright counterstaining. (J Histotechnol 12:210, 1989).     

Victoria Blue-Hematoxylin and Eosin Staining: A Useful Routine Stain for Demonstration of Venous Invasion by Cancer Cells

Abstract

Victoria blue (VB)-hematoxylin and eosin (H&E) staining, originally described by Yokokawa et al in 1983, has been used extensively in our surgical pathology lab as a routine stain for all surgical specimens with cancer. This simple staining sequence is useful for the demonstration of venous invasion by cancer cells as VB dye specifically demonstrates elastic fibers in the background of H&E-stained tissues. (The J Histotechnol 13:271, 1990)     

Modification of Brain Processing Techniques: Tissue Embedding in Glycol Methacrylate and Stain Modifications

Abstract

Increased interest in quantitation of the histopathological changes in a variety of neurological disorders (including neurodegenerative disease such as Alzheimer's disease) continues in an attempt to develop specific clinical-histopathological correlations. Most previous efforts at quantitation have used paraffin embedded sections of brain tissue, although plastic embedded sections have recently become a preferable alternative because they provide greatly reduced tissue shrinkage and distortion during processing, and greater clarity and improved resolution to the tissue sections. We have developed techniques for glycol methacrylate embedding and sectioning of brain tissue blocks on a standard histology laboratory microtome. In addition, we have modified routine diagnostic and investigational neurohistological stains for use in glycol methacrylate embedded brain sections, including hematoxylin and eosin, modified Bielschowsky stain, Jamarri silver technique, Einarson's Nissl stain, gallocyanin-Darrow red myelin stain, and the thioflavine-S-hematoxylin stain. The use of plastic embedded sections with appropriate stains will permit critical histopathological evaluation of nervous system tissue from patients with a variety of neurological disorders. (J Histotechnol 12:201, 1989)     

A Polychrome Stain for Mucopolysaccharides on Semithin Sections of Epoxy Resin Embedded Tissues

Abstract

The method describes a simple, quick polychromatic stain for Epon-Araldite and Araldite embedded tissue. It differentially stains mucins and mucopolysaccharides. The technique is simple to use and requires a hot plate to carry out the stain, which is done with droplets of Milipore filtered stain solution. It is useful as a screening stain or as a stain for some tissue components that generally require complex and prolonged procedures. A wide range of shades are produced, with the best effects achieved in tissues with abundant connective tissue, mucopolysaccharides, or granulocytic cells. Intraluminal bacteria of gastric biopsies are also demonstrated.     

Observations on Fixation and Staining of Bone Marrow Biopsies

Abstract

Ten bone marrow (BM) biopsies were taken from the iliac crest of six autopsy cases. The tissues were fured in Zenker's fixative for intervals between one and 18 hours with and without decalcification. Ultrasonic bombardment was used for one and two hours. The same procedures were also used following 18 hours fixation in 10% neutral buffered formalin. Coded slides were double blind evaluated for section quality and cytologic detail after staining with H & E, three Giemsa variants, and Gomori's iron stain.

Paraffin processed BM biopsies were optimally prepared by fixation in Zenker's fluid for 4-18 hours without decalcification. Prior formalin fixation did not influence the consistent brilliance of Giemsa staining with the preferred Sheehan Wright Giemsa stain. Overnight Zenker fixation with or without prior formalin exposure and the absence of decalcification is recommended for the routine preparation of BM biopsies for paraffin processing. (The J of Histotechnol 11:145, 1988.)     

A Combination Stain for Acid Fast Bacilli and Fungi

Abstract

A combination stain for acid fast bacilli and fungi is described. The method is simple and it provides a reliable method of recognizing both organisms in tissue sections simultaneously in one setting. Basically it is a combination of Beamer's modification of Fites for acid fast bacilli and Gridley's stain for fungi.     

Variation of the Holmes Method for Histologic Staining of Bone Canaliculi

Abstract

The Holmes method for soft tissue staining of nerve fibers was modified to produce excellent detail of the canalicular network in decalcified bone tissue. With simple changes in timing and the meticulous use of fresh reagents, we were able to stain bone lacunae and canaliculi in a variety of animal and human tissues. This technique produced more uniform and accurate results than the thionin/picric acid stains used previously for visualization of osteocytes. The morphologic results can be used for diagnostic purposes as well as stereological studies where the osteocyte network is of importance. (The J Histotechnol 16:355, 1993)     

Enzyme-linked immunofiltration assay for rapid serodiagnosis of syphilis

A new rapid technique for detection of serum treponemal antibodies is described which is based on an enzyme-linked immunoassay using nitrocellulose as solid phase. With this technique antigen-antibody binding is accelerated by the filtration of the antibody solution through the antigen-coated nitrocellulose filter instead of its remaining over the solid phase for incubation. Test results are available in less than 15 min. Serum specimens from 255 syphilitics and 829 non-infected subjects were investigated. The sensitivity and specificity of the Treponema pallidum enzyme-linked immunofiltration assay were comparable to those of the Treponema pallidum haemagglutination assay and the fluorescent treponemal antibody absorption test.     

Silver Labeling of Alzheimer Neurofibrillary Changes and Brain β-Amyloid

Abstract

Alzheimer neurofibrillary changes and brain β-amyloid can be stained selectively by silver impregnation on tissue sections and (in the case of neurofibrillary proteins) on sodium dodecyl sulfate-polyacrylamide gels. The stain can be applied to frozen sections of tissue from 10–150 μm thick, to similarly thick polyethylene glycol sections, or to 5–15 μm thick paraffin sections. The technique can also be applied to routinely fixed autopsy material. The technique takes advantage of the physical development of nucleation sites, thereby permitting tight control of the entire procedure. It is less expensive than immunocytochemical techniques, and facilitates processing of large numbers of sections through entire hemispheres of the human brain. (The J Histotechnol 16:335, 1993)     

Field's stain – a rapid staining method for Acanthamoeba spp.

Acanthamoeba sp. is a free-living amoeba known to cause chronic central nervous system infection or eye infection in humans. Many cases remain undetected for want of a good detection system. We report for the first time a rapid staining method to facilitate the identification of Acanthamoeba sp. using the modified Field's staining technique. A. castellanii, which was used in the present experiment, is maintained in our laboratory in mycological peptone medium (Gibco). The cultures were pooled together and smears were made on glass slides for staining purposes. Different types of stains such as Field's stain, modified Field's stain, Wright's stain, Giemsa stain, Ziehl-Neelsen stain, and trichrome stain were used to determine the best stain for the identification of this amoeba. The concentration of various stains and the duration of staining were varied to provide the best color and contrast for each stain. Acanthamoeba was also obtained from the brain of experimentally infected mice and was stained with various stains as mentioned above to determine the best stain for use in identifying the presence of this parasite in experimentally infected animals. The modified Field's stain gives a very good color contrast as compared with other stains. Furthermore, it takes only 20 s to be carried out using the least number of reagents, making it suitable for both laboratory and field use. Received: 24 March 1999 / Accepted: 16 April 1999     

Modified Specimen Holder for the Ames Cryostat

Abstract

An inexpensive device to adapt an Ames Cryostat is described. It utilizes a brass 25 mm specimen holder for frozen section technique. The adapter permits the use of specimen plates designed for other cryostats. It is inexpensive and simple to make from acrylic. (The J Histotechnol 13:297, 1990)     

Author and Title Index

Abstract

A rapid method of staining reticulin fibers in resin-embedded sections by ammoniacal silver nitrate is described. Microwaves are used to dry the tissue sections, to stimulate reaction with periodic acid, and to accelerate staining with silver nitrate. The procedure is rapid, simple and produces consistent results. (The J Histotechnol 12:289, 1989).     

Staining effect of Hibiscus sabdariffa extract on sperm cell morphology of Sprague–Dawley rats

Abstract

There is increasing awareness among people towards natural products. Due to their non-toxic properties, low pollution, and lower side effects, natural dyes are used in many day-to-day products. Although the African continent possesses plentiful plant resources, only a small amount has been exploited so far. This study evaluated the use of Hibiscus sabdariffa as a stain to evaluate sperm morphology. Following liquefaction, 10 μl of semen was spread onto glass slides and allowed to air-dry at room temperature. The smear was fixed for 15 minutes in methanol. The sperm morphology was analyzed by staining 10 slides of the smears with eosin (control) and the ethanolic extract of Hibiscus sabdariffa dye was used to stain the sperm cells. The smears were air-dried and viewed at magnification of ×400. Phytochemical and chromatographic analyses were carried out. The sperm cells were stained in shades of reddish brown. Preliminary phytochemical screening of Hibiscus sabdariffa revealed that it contains alkaloids, saponins, flavonoids, and tannins. Hibiscus sabdariffa has potential for use as a stain for study of sperm morphology.