Rosado M Manuela, Scarsella Marco, Cascioli Simona, Giorda Ezio, Carsetti Rita
Research Center Ospedale Pediatrico Bambino Gesù (IRCCS), Rome, Italy.
Methods Mol Biol. 2013;1032:45-57. doi: 10.1007/978-1-62703-496-8_4.
Lymphocyte characterization is primarily based on the differential expression of surface markers. In this context, flow-cytometry analysis (FACS) is an exceptional technique that not only allows the identification of B-cell subsets, but can also be used to evaluate cell function, activation, and division. Here, we will combine the use of FACS analysis and ELISA techniques to identify murine bone marrow and peripheral B-cell subsets. The main function of B cells, derived through a multistage differentiation process from precursor cells, is to produce antibodies. This task is performed by terminally differentiated B cells called antibody-secreting cells (ASC) present at mucosal sites, in the bone marrow and in the spleen. The number and specificity of ASC can be measured by Enzyme-linked immunosorbent spot (ELISPOT) assay, a variation of the enzyme-linked immunosorbent assay (ELISA) used to quantify serum immunoglobulins.
淋巴细胞的特征主要基于表面标志物的差异表达。在此背景下，流式细胞术分析（FACS）是一种卓越的技术，它不仅能够识别B细胞亚群，还可用于评估细胞功能、激活和分裂。在这里，我们将结合使用FACS分析和ELISA技术来鉴定小鼠骨髓和外周血B细胞亚群。B细胞通过从前体细胞进行多阶段分化过程产生，其主要功能是产生抗体。这项任务由终末分化的B细胞完成，这些细胞称为抗体分泌细胞（ASC），存在于黏膜部位、骨髓和脾脏中。ASC的数量和特异性可以通过酶联免疫斑点（ELISPOT）测定法来测量，这是一种用于定量血清免疫球蛋白的酶联免疫吸附测定（ELISA）的变体。
