Trichrome Staining Techniques Research Articles

Depth of tissue injury during culdotomy using the ultrasonically activated scalpel

The ultrasonically activated scalpel is an energy source which relies on mechanical, vibrational energy rather than thermal energy to incise tissue and achieve hemostasis. To determine the zone of tissue necrosis, microscopic sections were taken from the midpoint of culdotomy incisions performed during laparoscopic hysterectomy. Photomicrographs using trichrome staining technique indicate an approximately 1 mm zone of denaturation of collagen. Minimal coagulation necrosis or thermal artifact occurred. We conclude the ultrasonically activated scalpel may be used with safety in proximity to pelvic structures at risk for injury.

Evaluation of a commercially available enzyme-linked immunosorbent assay for Giardia lamblia antigen in stool

The lack of a quick, simple, and inexpensive diagnostic test has limited the ability of public health officials to rapidly assess and control outbreaks of Giardia lamblia in child day-care centers. We evaluated the performance of a commercially available enzyme-linked immunosorbent assay (ELISA) for the detection of a G. lamblia-associated antigen in stool. Stool specimens were collected from the diapers of 426 children attending 20 day-care centers, fixed in 10% Formalin and polyvinyl alcohol, and examined by microscopy by Formalin concentration and trichrome staining techniques. Specimens were also tested visually and spectrophotometrically by ELISA. Of 99 tests positive by microscopy, 93 were visually positive by ELISA (sensitivity, 93.9%). Of 534 tests negative for G. lamblia by microscopy, 32 (6.0%) were ELISA positive. However, on the basis of examination of multiple specimens from the same child, none of these could be considered false-positive ELISAs; the specificity of the ELISA was therefore 100%. The sensitivity of both microscopy and ELISA improved as the number of specimens per child increased. An optical density value of greater than 0.040 was 98.0% sensitive and 100% specific for G. lamblia. This ELISA, which appeared to be more sensitive for G. lamblia than did microscopic examination of stool, should be useful as an epidemiologic tool, particularly in day-care settings, and may also have a role in confirming clinical diagnoses of giardiasis.

Use of terpene-based products, with and without anhydrous alcohol, as clearing agents in trichrome staining technique for intestinal protozoa

BDH xylene substitute, a terpene-based product, and its mixture with anhydrous alcohol were found to be excellent replacements for xylene and carbol-xylene, respectively, in the trichrome staining technique applied to sodium acetate-acetic acid-Formalin-fixed fecal smears for detection of intestinal protozoa.

Differential Staining of the Cell Cycle of Plant Cells Using Safranin and Indigo-Picrocarmine

A trichrome staining technique using safranin-indigo-picrocarmine (SIPC) can be used to distinguish the various stages of the cell cycle in onion root tip. When the tissue was fixed first in formalin followed by picric acid and stained in SIPC, a clear differentiation of interphase nuclei into four color classes, viz., green, orange, red and brown can be recorded. Replacing crystal violet for safranin produces a similar pattern of differentiation of interphase nuclei into green, light blue, blue and deep blue. Autoradiographic study using 3H-thymidine as a DNA precursor demonstrates the reliability of the SIPC staining technique. All the orange and red nuclei are found to be labelled and therefore are in S phase of the cell cycle. Almost all the green nuclei are unlabelled and may be assigned to G1. The larger brown nuclei which are mostly unlabelled can be considered in G2 phase.

Modified trichrome staining technique with a xylene substitute.

Hemo-De was evaluated and found to be a suitable replacement for xylene in the Wheatly trichrome staining of polyvinyl alcohol-fixed fecal smears used in the microscopic identification of intestinal protozoans.

Detection of helminth ova and larvae in trichrome-stained stool smears.

The detection and identification of intestinal helminths were studied retrospectively by comparing the Formalin concentration technique with the trichrome-stained smear technique. A total of 3,997 stool samples from 1,570 patients were examined by both methods. Of the 3,997 samples, 31% (1,239 of 3,997) contained helminths or protozoans or both. A total of 11% (434 of 3,997) of the samples representing 14% (221 of 1,570) of the patients were positive for one or more helminth species. A total of 570 separate identifications of helminth ova/larvae were made. Among the helminth ova/larvae identified. 14.6% (83 of 570) were detected only in the trichrome-stained smear, representing 6.3% (14 of 221) of the patients. From these data, it can be concluded that unless a diligent search of the stained smear for helminths is made, a significant number of helminth infections may be missed. It is, therefore, recommended that stained stool smears be used to aid the detection of helminth ova/larvae in conjunction with Formalin concentration. The appearance of the most common helminth ova/larvae in trichrome-stained smears is described, along with specific characteristics that may be used for their identification. In addition, the advantages and disadvantages of the trichrome staining technique for helminth identification are discussed.

Detection of different functional states of the cellular nucleus with a new trichrome staining technique

The new trichrome stain ASTRIN, prepared from alcian blue, safranine, thionine, and resorcin, stains the cell nuclei red or blue, depending on their functional state. Activation of salivary gland cells with pilocarpin, or of liver cell with prednisolone, equally resulted in a quantitative increase of cells with active (blue) nuclei. Changes in nuclear staining after digestion with trypsin, pronase, or ribonuclease, and after extraction of cellular DNA and RNA with perchloric acid, unequivocally suggest that alteration of the protein-DNA ratio, i.e. the hetero- or euchromatic state of nuclear chromatin, is responsible for the disparity of the nuclear staining reaction with ASTRIN. The new trichrome stain seems to be suitable for routine cytodiagnostic use.

An amphoteric character in mast cell granules of Amphisbaenia fuliginosa, a snakelike lizard.

By the use of haematoxylin-eosin and Masson's trichrome staining techniques, and of histochemical tests with Alcian blue and metachromasia with toluidine blue, the mast cell granules of the snakelike lizard Amphisbaenia fuliginosa are shown to present an amphoteric character: they are stained by the acid as well by the basic dyes.This property is here discussed in function of the literature, with the main concern directed toward the taxonomic position of this animal. The Amphisbaenidae seem to represent an intermediate stage between lizards and ophidians.The mast cell granules of lizards are basophilic and metachromatic; in the ophidian, typical mast cells have not been evidenced yet, but a granular acidophilic cell has been demonstrated in the connective tissue. The basophilic and acidophilic characters of these two cells seem to be intermingled in the mast cell of the Amphisbaenidae.