Schlegel R A, Stevens M, Lumley-Sapanski K, Williamson P
Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Immunol Lett. 1993 Jun;36(3):283-8. doi: 10.1016/0165-2478(93)90101-7.
To test whether apoptotic thymocytes can be identified by altered packing of lipids in their plasma membranes, thymocytes were isolated from mice injected with hydrocortisone and stained with merocyanine 540 (MC540), a fluorescent probe sensitive to lipid packing. At 9-10 h after injection, a subpopulation of H2-Klo cells with increased MC540 staining was clearly discernable. When DNA degradation was assessed by staining fixed cells with propidium iodide (PI), the fraction of cells with increased MC540 staining corresponded to the fraction with reduced PI staining. In addition, enriching for the former by fluorescence-activated cell sorting enriched for the latter. These results indicate that living apoptotic thymocytes can be identified and separated on the basis of altered lipid packing and increased staining with MC540.
为了检测凋亡胸腺细胞是否可通过其质膜中脂质堆积的改变来识别，从注射了氢化可的松的小鼠中分离出胸腺细胞，并用部花青540（MC540）染色，MC540是一种对脂质堆积敏感的荧光探针。注射后9 - 10小时，可明显辨别出MC540染色增加的H2-Klo细胞亚群。当用碘化丙啶（PI）对固定细胞染色来评估DNA降解时，MC540染色增加的细胞比例与PI染色减少的细胞比例相对应。此外，通过荧光激活细胞分选富集前者也会富集后者。这些结果表明，活的凋亡胸腺细胞可基于脂质堆积改变和MC540染色增加来识别和分离。
