The Acridine Orange Viability Test Applied to Bone Marrow Cells I. Correlation with Trypan Blue and Eosin Dye Exclusion and Tissue Culture Transformation

Hathaway, William E.; Newby, L.A.; Githens, John H.

doi:10.1182/blood.v23.4.517.517

Cited by 49 publications

(18 citation statements)

References 13 publications

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“…for 468.3603). The 1 H-and 13 methylenes, two methines, and four quaternary alkane carbon atoms. Comparison of these data with those of related, known lanostane triterpenoids revealed that 1 is a 24-methyl-5α-lanostane containing a Δ 25 double bond [9].…”

mentioning

confidence: 99%

Antibacterial Compounds from Mushrooms II: Lanostane Triterpenoids and an Ergostane Steroid with Activity AgainstBacillus cereusIsolated fromFomitopsis pinicola

et al. 2009

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Anti- Bacillus cereus bioassay-guided fractionation of a crude extract of the American mushroom, Fomitopsis pinicola, was performed using thin-layer chromatography, Sephadex LH-20 column chromatography, and preparative-scale HPLC. Five lanostane triterpenoids (1-5) and one ergostane steroid (6) were isolated and identified. Compound 1 is a new lanostane triterpenoid, and its structure was determined using 1D and 2D NMR experiments, HR-MS, and physical data. Each of the purified compounds (1-6) was tested for antibacterial activity against B. cereus using standard MIC assays. Compounds 1-6 had MIC values of 32, 16, 32, 32, 128, and 64 microg/mL, respectively.

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confidence: 99%

Antibacterial Compounds from Mushrooms II: Lanostane Triterpenoids and an Ergostane Steroid with Activity AgainstBacillus cereusIsolated fromFomitopsis pinicola

et al. 2009

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“…o cover glass which is then ringed with melted paraffin. Viability counts are made with a Zeiss fluorescence microscope according to the method of Hathaway, Newby and Githens (1964).…”

Section: Methodsmentioning

confidence: 99%

Separation of Lymphocytes of Rat Bone Marrow by Combined Glass‐Wool Filtration and. Dextran‐Gradient Centrifugation

Morrison¹

1967

Br J Haematol

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SUMMARY The two‐step separation of rat bone marrow involving glass‐wool filtration combined with dextran‐gradient centrifugation yields viable cell suspensions composed predominantly of lymphocytes with a minimum of contamination by granulocytes (less than 10 per cent). Because filtration of rat bone marrow through glass wool removes the larger marrow cells and most of the immature forms of the red cell series, it is useful as a first step in the separation of lymphocytes from bone marrow. Density‐gradient centrifugation of the glass‐wool filtrate serves to differentially settle out lymphocytes and granulocytes when dextran solutions of appropriate densities are used. The middle or layer 2 (density 1.0600) of the dextran gradient yields the greatest absolute numbers of lymphocytes. Small numbers of transitional and blast cells are also present in this layer. The separation procedure results in excellent preservation of cell morphology as evidenced by Jenner‐Giemsa staining. A high level of viability of the separated cells is indicated by dye exclusion, characteristic fluorescence in the presence of acridine orange, and capacity for oxygen consumption. In vivo viability of the separated cells is also indicated by their capacity to confer protection following their infusion into lethally irradiated rats.

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“…TC 199, pH 7.2) had been added. The numbers and differential counts of cells showing yellow-green nuclear fluorescence as a sign of cell vitality (Hathaway et al 1964) were determined by counting the cells in one or several transverse sectors in the chambers, using a Leitz Laborlux microscope with a x 40 objective and an Osram mercury Vapor lamps as the light source. A BG 12 blue exciter filter was used with a yellow-orange absorption filter.…”

Section: Microscope Slide Paraffin Sheet Coverslipmentioning

confidence: 99%

A Micro‐Incubation Chamber Technique for Testing the Immunological Activity of Cultured Lymphocytes

1968

Scandinavian Journal of Haematology

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The micro‐incubation chamber technique of Cunningham (1965) has been used for testing transformed cells in sheep red cell stimulated lymphocyte cultures for the ability to give pericellular lysis (lytic plaques) in sheep red cell monolayers. The variations around the means for total cell numbers and plaque‐forming cell numbers in series of chambers plated in parallel have been estimated. The plaque‐forming ability was found to be dependent on complement and temperature indicating an immunological lytic reaction. Evidence is presented for active protein synthesis in and liberation from the cells showing the ability to form lytic plaques. Cells stored at low temperature were able to form lytic plaques after several days' storage although a gradual reduction in the plaque‐forming ability was noticed.

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The Acridine Orange Viability Test Applied to Bone Marrow Cells I. Correlation with Trypan Blue and Eosin Dye Exclusion and Tissue Culture Transformation

Cited by 49 publications

References 13 publications

Antibacterial Compounds from Mushrooms II: Lanostane Triterpenoids and an Ergostane Steroid with Activity AgainstBacillus cereusIsolated fromFomitopsis pinicola

Antibacterial Compounds from Mushrooms II: Lanostane Triterpenoids and an Ergostane Steroid with Activity AgainstBacillus cereusIsolated fromFomitopsis pinicola

Separation of Lymphocytes of Rat Bone Marrow by Combined Glass‐Wool Filtration and. Dextran‐Gradient Centrifugation

A Micro‐Incubation Chamber Technique for Testing the Immunological Activity of Cultured Lymphocytes

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