Differential analysis of animal bone marrow by flow cytometry

Martin, Ronald A.; Brott, David; Zandee, J.C.; McKeel, Mildred J.

doi:10.1002/cyto.990130612

Cited by 23 publications

(17 citation statements)

References 8 publications

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“…By sorting analysis, the populations of E, G, M and L&T were composed of erythrocytes, granulocytes, monocytes, and a mixture of lymphocytes plus thrombocytes, respectively as described previously [4]. Figure 2A show the analysis of the same blood sample after staining with DiOC 5 (3) instead of DiOC 6 (3). In this case, FL-1 was linearly obtained.…”

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Counting Absolute Number of Lymphocytes in Quail Whole Blood by Flow Cytometry

Uchiyama

Moritomo

Kai

et al. 2005

The Journal of Veterinary Medical Science

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ABSTRACT. In a previous study, we reported a new method for counting quail blood cells. After quail blood cells were stained with fluorescent lipophilic dye (DiOC 6 (3)), absolute counts of erythrocytes, granulocytes, and monocytes were obtained by means of flow cytometry (FC). The FC method has the potential for application to avian blood cells count; however, the method was unable to distinguish between lymphocytes and thrombocytes. In the present study, we improved the FC method to obtain separate counts of lymphocytes using DiOC 5 (3). After quail blood cells were stained with DiOC 5 (3), the cells were measured with FC. Each blood cell type was distinguished by means of their typical FL-1 (green fluorescence) and SSC (side scatter). Absolute numbers of erythrocytes, granulocytes, monocytes and lymphocytes in whole blood were obtained. The improved FC analysis worked equally well with chicken (Gallus gallus) and goose (Anser cygnoides) blood. KEY WORDS: avian, flow cytometry, lymphocyte count.

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“…5A and B). DiOC has been used in several applications, such as differentiation of bone marrow cells of dogs [3], counting of reticulocytes in humans [7], and counting of blood cells of fish [1]. This agent was used here to facilitate the resolution of different cell lineages in avian leukocytes.…”

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Counting Absolute Number of Lymphocytes in Quail Whole Blood by Flow Cytometry

Uchiyama

Moritomo

Kai

et al. 2005

The Journal of Veterinary Medical Science

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“…DiOC 6 (3) is known to be cationic and lipophilic dye that selectively stain lipid bilayers such as the plasma membrane and cell organelle membranes [6,7]. The DiOC 6 (3) was good at staining leukocytes and thrombocytes, but not as effective at staining erythrocytes, since erythrocytes had poor cell organelle [3,8]. In the present study, we applied DiOC 6 (3) staining to quail (Cotumix cotumix japonica) blood cells to establish a new method for counting avian blood cells.…”

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A New Method for Counting of Quail Leukocytes by Flow Cytometry.

et al. 2002

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ABSTRACT. An automatic counting method was developed for fish blood cells using a fluorescent dye, 3, 3-dihexyloxacarbocyanine (DiOC 6 (3)), that selectively stain lipid bilayers in living cells. In the present study, the DiOC 6 (3) method was applied to quail (Cotumix cotumix japonica) blood cells. After quail blood cells were stained with DiOC 6 (3), absolute counts and relative proportions of erythrocytes, granulocytes, monocytes, and lymphocytes plus thrombocytes in whole blood were obtained by means of flow cytometry (FC). The number of each cell types by the FC was in good agreement with those counted microscopically. This method will offer new possibilities for routine blood cell counting for avian medicine. KEY WORDS: avian, flow cytometry, leukocyte count.

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“…The lipophilic cationic dye 3,3Ј-dihexyloxacarbocyanine iodide has been used in studies of human red cells (17), murine bone marrow (13), and to differentiate bone marrow in multiple species (11). Although this method for performing differentials proved useful, categories of hematopoietic cells were frequently close to one another or overlapped, making it difficult to train technologists to divide the populations readily and reproducibly.…”

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Comparison of flow cytometric and manual bone marrow differentials in Wistar rats

Criswell¹,

Bleavins²,

Zielinski³

et al. 1998

Cytometry

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Preclinical drug trials frequently require the evaluation of animal bone marrow, a time‐consuming process requiring the skills of a highly trained hematologist. In the present study, a flow cytometric technique was developed that could effectively replace the need for manual bone marrow differentials in rats. Peroxidase activity, measured indirectly with 2′7′‐dichlorofluorescin, was coupled with the use of species‐specific T‐ and B‐lymphocyte antibodies and cell size to produce a flow cytometric analysis of rat bone marrow. Accurate identification of lymphocyte, proliferating and maturing erythroid and myeloid, and megakaryocyte populations was confirmed by cell sorting. Flow cytometry yielded differentials that were indistinguishable from manual differentials and published reference ranges. Enumeration of lymphocyte numbers with monoclonal markers is a key advantage of flow cytometric differentials because misidentification of lymphocytes in poorly prepared or stained bone marrow smears is a common problem. The most apparent advantage is increased throughput and reproducibility. Operator training for analysis using flow cytometry can be readily accomplished within a few days as opposed to the extensive training required for individuals performing manual bone marrow differentials. This methodology provides a high‐volume, rapid, and relatively low‐cost tool for the reliable evaluation of rat bone marrow differentials that has been heretofore unavailable. Cytometry 32:9–17, 1998. © 1998 Wiley‐Liss, Inc.

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Differential analysis of animal bone marrow by flow cytometry

Cited by 23 publications

References 8 publications

Counting Absolute Number of Lymphocytes in Quail Whole Blood by Flow Cytometry

Counting Absolute Number of Lymphocytes in Quail Whole Blood by Flow Cytometry

A New Method for Counting of Quail Leukocytes by Flow Cytometry.

Comparison of flow cytometric and manual bone marrow differentials in Wistar rats

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