Determining the abundance of leukocyte subtypes, including lymphocyte subpopulations, not only in blood but also in lymphatic tissues, is inevitable to assess the immune status of an organism for research purposes. However, nucleated thrombocytes and erythrocytes exacerbate many hematological techniques in avian species. In order to enable a rapid discrimination of leukocyte subsets from lymphatic tissues of chicken, we adapted existing flow cytometric methods for counting leukocytes in chicken blood. We established staining and gating strategies allowing the flow cytometric characterization and enumeration of total leukocytes, thrombocytes, monocytes/macrophages, CD8α⁺ lymphocytes, CD4⁺ T cells, γδ T cells, and B cells in chicken spleen and CD8α⁺ lymphocytes, CD4⁺ T cells, γδ T cells, and B cells among intraepithelial lymphocytes in chicken cecal tonsils. For this, we prepared single‐cell suspensions of spleen and isolated intraepithelial lymphocytes from cecal tonsils without density centrifugation, and performed antibody staining of cells without subsequent washing steps to prevent cell loss and falsification of obtained cell counts. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.

中文翻译：

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通过流式细胞术表征来自淋巴组织的鸡白细胞亚群

确定白细胞亚型的丰度，包括淋巴细胞亚群，不仅在血液中，而且在淋巴组织中，对于出于研究目的评估生物体的免疫状态是不可避免的。然而，有核血小板和红细胞加剧了禽类中的许多血液学技术。为了能够快速区分鸡淋巴组织中的白细胞亚群，我们采用了现有的流式细胞术方法来计算鸡血中的白细胞。我们建立了染色和门控策略，允许对鸡脾和 CD8α ^{+ 中}的总白细胞、血小板、单核细胞/巨噬细胞、CD8α ⁺淋巴细胞、CD4 ⁺ T 细胞、γδ T 细胞和 B 细胞进行流式细胞术表征和计数鸡盲肠扁桃体上皮内淋巴细胞中的淋巴细胞、CD4 ⁺ T 细胞、γδ T 细胞和 B 细胞。为此，我们在没有密度离心的情况下制备了脾脏和从盲肠扁桃体中分离的上皮内淋巴细胞的单细胞悬液，并在没有后续洗涤步骤的情况下对细胞进行了抗体染色，以防止细胞丢失和获得的细胞计数造假。© 2020 作者。细胞学 A 部分由 Wiley Periodicals LLC 代表国际细胞学促进会出版。