Evaluation of a Cytocentrifuge Technique Effects on Quantitative Determination of Feulgen-Deoxyribonucleic Acid and Total Dry Mass of Human Leukemic Cells

Bault, Lawrence E. De

doi:10.1177/20.9.703

J Histochem Cytochem.

1972

DOI: 10.1177/20.9.703

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Evaluation of a Cytocentrifuge Technique Effects on Quantitative Determination of Feulgen-Deoxyribonucleic Acid and Total Dry Mass of Human Leukemic Cells

Lawrence E. De Bault

Abstract: The cytocentrifuge has been used as a tool to obtain preparations from suspension cultures suitable for quantitative cytochemical analysis.

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1976

1981

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A new type of cytocentrifuge, the valve-centrifuge

1977

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A new type of cytocentrifuge has been developed in which the sedimentation process of the cells onto the slides is separated from the draining of the sedimentation fluid. This is realised by electrically controlled valves which can be closed and opened while the centrifuge is running. Sedimentation is carried out with closed valves, draining of adhering medium with open valves. The preparations, freed of adhering medium by the centrifugal force can be taken out and the cells can be fixed. Alternatively the valves can be closed again and fixative can be introduced through a central well, the cells still being under the influence of the centrifugal force. With subsequent draining of the fixative and introduction of washing and staining solutions through the central well, the whole process from sedimentation to staining can be carried out in the running centrifuge. The process seems well suited for complete automation. Using dilution series from a suspension of human buffy coat cells counted in a Buerker chamber, the cell counts in the centrifuge preparations showed virtually total recovery of cells, with no apparent selection or specific distribution of cell types. Draining of the sedimentation and fixative fluids at a slow rate was found to be vital for optimal recovery of cells. The morphology of different cell types sedimented on the slide was excellent. The flattening of nuclei thorugh gravity was studied by cytophotometry of Feulgen-stained leucocytes. The nuclear area of these cells was found to be approximately double that from cells in identically stained classical smears. With this type of valve-centrifuge a quantitative and unbiased recovery of uniformly spread and flattened cells on coverslips or slides may be obtained, thus making the procedure well suited to automated analysis based on cytophotometric principles and morphometric pattern recognition.

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A new type of cytocentrifuge, the valve-centrifuge

1977

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Quantification of nuclear non-histone proteins by Feulgen-Naphthol Yellow S cytophotometry

Gaub

1981

Histochem J

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Owing to the accumulation of nuclear non-histone protein (NHP) (a) in cells entering the cell cycle from the quiescent state and (b) in continuously cycling cells during G1 phase, a simultaneous determination of DNA and nuclear NHP is of high potential utility in cell kinetic studies. This paper provides guidelines for a Feulgen--Naphthol Yellow S staining technique for this purpose. It discusses details of the preparation and quantification procedures, and reviews the evidence for a quantitative relationship between nuclear Naphthol Yellow S binding and nuclear NHP.

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The detection of cytoplasmic immunoglobulins by immunofluorescence: Improvements in techniques and standardization procedures

Vossen

Bergh

1976

Journal of Immunological Methods

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Evaluation of a Cytocentrifuge Technique Effects on Quantitative Determination of Feulgen-Deoxyribonucleic Acid and Total Dry Mass of Human Leukemic Cells

Abstract: The cytocentrifuge has been used as a tool to obtain preparations from suspension cultures suitable for quantitative cytochemical analysis.

Cited by 3 publications

References 9 publications

A new type of cytocentrifuge, the valve-centrifuge

A new type of cytocentrifuge, the valve-centrifuge

Quantification of nuclear non-histone proteins by Feulgen-Naphthol Yellow S cytophotometry

The detection of cytoplasmic immunoglobulins by immunofluorescence: Improvements in techniques and standardization procedures

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