May-Grünwald-Giemsa Stain Research Articles

North Carolina Turkey Enteric Virus: Further Characterization

SUMMARY The North Carolina turkey enteric virus was further characterized on the basis of certain physical and chemical properties. Intracytoplasmic staining of infected turkey kidney cells was demonstrated by fluorescein-labeled antiserum. May-GrunwaldGiemsa staining revealed very large syncytia. An intense greenish color was observed in syncytia stained with acridine orange. The virus was relatively heat-stable and pH-stable, and plaques of slightly increased size were produced in the presence of magnesium chloride. The virus did not hemagglutinate when a number of avian and mammalian red blood cells were employed. The growth curve was typical of reoviruses. Pathogenicity for 13-day-old embryonated turkey eggs was demonstrated.

Cells in bovine milk: differential staining in suspension: collecting, counting and examining on millipore membrane.

Differential staining of somatic cells in bovine milk preserved with 12 mg of HgCl2 and 52 mg of K2Cr2O7 in 50 ml was performed as follows. (1) To a silicone-coated serological test tube are added 2 ml of 0.1% Triton X-100 solution, pH 7.0 (prepared with distilled water and filtered through a membrane filter), and 0.16 ml of benzidine-H2O2 solution (benzidine 0.2 gr/200 ml distilled water, filtered, to which are added 0.2 ml of 3% H2O2). (2) A 0.2 ml sample of milk is added, mixed well and the tube allowed to stand for 3 min at room-temperature. (3) 0.025 ml of May-Grunwald stain (British Drug Houses standard stain solution, No. 16934/3097) and 0.08 ml of Giemsa stain (improved Giemsa stain G. T. Gurr R66, in solution; BDH Cat. No. 17401) are added, mixed well and allowed to stand for 20 min in a water bath at 48-50 C. The contents and two rinsings of the tube with Triton solution at 48-50 C are poured into a siliconized Pyrex microanalysis filter holder (Millipore XX10 02500) with a Millipore (25 mm dia....

The Use of Pyrogallol to Demonstrate Peroxidase in Mammalian Blood Eosinophils

Canine blood films were fixed in a mixture of formaline (40% HCHO) and 95% ethyl alcohol, 1:9, for 30 sec, washed in distilled water and air dried. A mixture of 10% aqueous pyrogallol 6 ml and H2O2 (6% or 20 vols.) 0.1 ml were applied to the film, allowed to react for 6 min and then washed off with distilled water. The film was counterstained with May-Grunwald Giemsa, Leishman, or Giemsa stain. This method stains canine eosinophils specifically for the presence of peroxidase, but has variable effects on eosinophils of other mammalian species, depending on the type of fixative used. Modified techniques are described for 4 other mammalian species and the possible causes of the staining variations are discussed.

The ingestion of bacteria and of altered bacterial structures by Entamoeba histolytica in Shaffer-Frye and in modified Shaffer-Frye media.

Summary Three cultural variants of the Gram-negative anaerobic bacillus used in the Shaffer-Frye (S-F) medium for propagation of Entamoeba histolytica were studied in the S-F medium and in the modified S-F medium of Reeves, et al. Culture S1, which is now used routinely in the S-F medium was found to produce round bodies, probably protoplasts, under the influence of the penicillin G in both media. These bodies were ingested by the amebae during their propagation. At no time were ingested bacterial cells seen with culture S1. Culture LSU, which is used routinely in the modified S-F medium, developed considerable cellular granulation under the influence of the penicillin G, but did not form round bodies in either medium. With this culture, bacterial cells were ingested by the amebae during their propagation. Culture S6, which does not support propagation of E. histolytica in either medium, also did not form round bodies in either medium. Nevertheless, the amebae ingested the bacterial cells of this culture. Darkfield microscopy was used for all direct observations in these studies. Permanent preparations were made by fixation of the cultures with 10% formalin followed by staining with the May-Grunwald stain. Within the limits of these experiments, it appears that the observed changes induced by penicillin G were inherent characteristics of the bacterial cultures and not the result of variation between the S-F and the modified S-F media. The fact that the ameba ingested bacterial cells (LSU and S6) or round bodies (S1) regardless of whether propagation was supported or not, was interpreted as an indication that ingestion of a particle does not necessarily imply that the particle is useful to the ameba.

Leukemia in the F Strain of Mice: Observations on Cytology, General Morphology, and Transmission

While many strains of mice with a high incidence of mammary tumors have been studied, only one strain with a comparable incidence of leukemia has been described (1). Stocks3 with a frequent appearance of leukemia have, however, been reported (2). The present communication concerns the incidence morphology and transplantation of leukemia in a second highly inbred pedigreed strain, the F strain, in which this disease occurs frequently. Materials and Methods The F strain of mice has been inbred (3), brother to sister, for a period of twelve years, for thirty generations. In the F4 and subsequent generations enlarged spleens, lymph nodes, and thymus glands have been observed in over 200 mice. Of the last 22 animals which were sacrificed or died at over six months of age, 10 had either leukemia or mediastinal lymphosarcoma. The present study is based on the last 17 spontaneous cases of leukemia and lymphosarcoma which appeared in this strain, plus 295 cases of leukemia developing in F mice following leukemic cell inoculations (Table I). Old, non-breeding mice of the F strain were examined twice a week to determine whether spleens and lymph nodes were enlarged. If these organs were found upon digital palpation to be larger than normal, total white blood cell and differential counts were made, the blood being obtained by cutting off the end of the tail and “milking.” The cover-slip method of making blood smears was used throughout. Animals were killed by severing the brain from the spinal cord. Dry imprints of spleen, lymph nodes, liver, bone marrow, and thymus were made. The freshly cut surface of the tissue was touched lightly to a clean slide, the imprinted material being waved dry immediately. On this type of preparation Pappenheim's May-Grunwald Giemsa staining combination was used. The May-Grunwald stain was allowed to remain on the slide for two minutes. An equal number of drops of distilled water buffered to a pH of 6.4 were then added to the undiluted stain and the diluted stain was allowed to remain on the slide until four minutes had elapsed from the beginning of the staining process. The stain was then poured off (no rinsing) and Giemsa solution (2 drops of stock solution to 1 c.c. of buffered distilled water) was poured on the slide. After fifteen minutes the slide was de-stained by pipetting several washings of buffered water over it. Blood smears were stained by the Giemsa solution for only ten minutes. For the peroxidase reaction Richter's method (4) was used. Following the staining process smears and imprints were air-dried, dipped in xylol, and mounted in damar.