Volume Distribution and Separation of Normal Human Leucocytes.

Dilla, M. A. Van; Fulwyler, Mack J.; Boone, Irene U.

doi:10.3181/00379727-125-32093

Cited by 53 publications

(17 citation statements)

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“…The high speed aind convenience of the systems allow statistically significant populations of cells to be analyzed, one cell at a time, on the basis of various parameters. These include measurements of cell volume to differentiate human leukocytes (1), the use of nuclear fluorochromes to determine differences in DNA content of chemically induced mouse tumor cells and normal cells (2,3), measurements of ultraviolet light absorption and light scatter to separate abnormal cells from uterine cervical carcinomas (4), and the measurement of nuclear to cytoplasmic ratios to detect malignant cells in gynecological specimens (5,6).…”

mentioning

confidence: 99%

Quantitative determination of transformed cells in a mixed population by stimultaneous fluorescence analysis of cell surface and DNA an individual cells.

Hawkes¹,

Bartholomew²

1977

Proc. Natl. Acad. Sci. U.S.A.

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ABSTACT Cell-surface labeling with fluorescamine indicates that the fluorescence of Balb 3T3 A31 cells is considerably decreased after both viral and chemical transformation. This phenomenon, coupled with the technique of flow microfluorometry, enabled nontransformed and transformed cells to be distinguished. A Second fluorescent probe, propidium iodide, which intercalatesinto DNA, was used in combination with fluorescamine in order to obtain a ratio of cell-surface labeling to DNA content: This manipulation allowed enhanced resolution of the two op tions'and the detection of small numbers of transfor abts n tr~dominantly normal population.

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mentioning

confidence: 99%

Quantitative determination of transformed cells in a mixed population by stimultaneous fluorescence analysis of cell surface and DNA an individual cells.

Hawkes¹,

Bartholomew²

1977

Proc. Natl. Acad. Sci. U.S.A.

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“…Flow cytometric differential blood cell counting is based on various principles, including cytochemical stains (9,15), fluorescent stains such as acridine orange (1,2,18), and differences in electronic cell volume (19) or in orthogonal light scattering (7,8). Differences in P12 uptake, demonstrated in the present report, provide a potential additional parameter for differential cell counting.…”

Section: Resultsmentioning

confidence: 69%

Flow cytofluorometric analysis of the uptake of the fluorescent fatty acid pyrene‐dodecanoic acid by human peripheral blood cells

et al. 1988

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The fluorescence activated cell sorter (FACS) was used for measuring the uptake of the fluorescent fatty acid derivative U-(l-pyrene) dodecanoic acid (Pa) by human peripheral blood cells. The results indicate that blood cells differ widely in their ability to take up Pl2, with polymorphonuclear cells showing the greatest uptake, followed by lymphocytes, platelets, and RBCs. These differences in Pl2 uptake provide a potential additional parameter for differential cell counting. Using the ability of the FACS to "gate out" nonrelevant cells, it was possible to measure the rate of Pl2 uptake by each respective cell type even when admixed with other cells. Thus elaborate physical separation procedures could be avoided, and contaminating cells did not influence the results. Differences in Pl2 uptake were also utilized to separate blood cells into pure subpopulations of specific cell types. Key terms: Fluorescence activated cell sorter, fatty acid derivativesA major obstacle in studying biological and biochemical properties of specific cells is the fact that, in vivo, cells are admixed with a variety of other cell types. For example, the study of hemopoietic cells is limited by the heterogeneity of a population consisting of cells belonging to several lineages that are at various stages of maturation. Analysis of such cells requires complete separation into discrete, homogenous subpopulations, since even a small contamination by cells that differ considerably in the property examined from the subpopulation studied may affect the results.We have demonstrated that the fluorescent, mediumchain fatty acid lZ-(l-pyrene) dodecanoic acid (P12) is effectively transported across the membranes of several cultured cell types and incorporated into their neutrallipids and phospholipids (10,ll). Using in vitro established leukemic cell lines, we have shown that cells of different hemopoietic lineages and stages of maturation differ in the rate and extent of PI2 uptake as well as in their metabolism of this acid (43). Recently, we have shown that the cellular uptake of P12 can be measured by the fluorescence activated cell sorter (FACS) (13,141, The present study describes the uptake of P12 by various cell types present in human peripheral blood and shows that blood cells differ widely in their ability to take up the compound. By taking advantage of the abilpossible to measure the rate of P12 uptake by each respective cell type, even when admixed with other cells. Thus elaborate separation procedures were avoided, and contaminating cells did not influence the results. Differences in P12-uptake were also utilized to separate blood cells into pure subpopulations of specific cell types. MATERIALS AND METHODS CellsPeripheral blood from normal individuals cells was collected in preservative-free heparin or citrate. Buffy coat cells were separated by mixing 2 volumes of blood with 1 volume of 1% dextran in saline (Pharmacia, Uppsala, Sweden) and allowing it to sediment at unit gravity for about 1 hour at room temperature. The cellcontai...

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“…The portion of the separation profile that erythrocytes and leukocytes did not separate very well was likely to be caused by the size overlap between erythrocytes and small lymphocytes. Although the volume of average small lymphocytes is at least twice that of erythrocytes [48], [49], the size is the determinant factor for the separation inside this device. The diameters of erythrocytes are reported between 7 and 9 µm, whereas those of small lymphocytes are between 6 and 9 µm [44], [46], [47].…”

Section: Resultsmentioning

confidence: 99%

Streamline-Based Microfluidic Devices for Erythrocytes and Leukocytes Separation

Zheng

Liu

Tai

2008

J. Microelectromech. Syst.

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In this paper, we report two devices for the continuous size-based separation of particles, such as blood cells, which is an important step for on-chip blood preparation. Unlike previously demonstrated passive fluidic devices for particle separation, the local geometry of the bifurcated side channels was used as a design parameter. The design of the devices was based on 2-D fluidic simulation of a T-shaped model. This novel approach was proved to be effective in predicting device performance. The critical particle size for separation was clearly defined in the bifurcated region by simulation under the established theoretical framework. We validated the operation principle of the devices by separating 5-and 10-µm polystyrene beads. Human leukocytes were also successfully separated from erythrocytes with 97% efficiency. The separation region of the device had a small footprint for the separation of particles in micrometer range, which makes this device a good candidate to be integrated into a lab-on-a-chip system. The particles were collected in different exit channels after they were separated, which facilitated further sensing and processing. Similar to cross-flow filters, particles were separated perpendicular to the flow direction. The filtering effect was achieved with the collection zones established by the fluidic field. Clogging was minimized by designing the minimal channel width of the devices larger than the largest particle diameter. Solvent exchange could be accomplished for particles.

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Volume Distribution and Separation of Normal Human Leucocytes.

Cited by 53 publications

References 0 publications

Quantitative determination of transformed cells in a mixed population by stimultaneous fluorescence analysis of cell surface and DNA an individual cells.

Quantitative determination of transformed cells in a mixed population by stimultaneous fluorescence analysis of cell surface and DNA an individual cells.

Flow cytofluorometric analysis of the uptake of the fluorescent fatty acid pyrene‐dodecanoic acid by human peripheral blood cells

Streamline-Based Microfluidic Devices for Erythrocytes and Leukocytes Separation

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