Outterridge P M, Jones W O, Edgar J A
CSIRO, McMaster Laboratory, Glebe, N.S.W., Australia.
Vet Immunol Immunopathol. 1988 Sep;19(2):141-51. doi: 10.1016/0165-2427(88)90005-0.
Sheep blood lymphocytes were labelled with fluorescent probes and examined under the fluorescence microscope and by the fluorescence-activated cell sorter. A novel probe using fluorescamine, coupled to hexylamine, detected 22.9% of cells, apparently of the B-cell series, counted by fluorescence microscopy. Substitution of fluorescein isothiocyanate (FITC) for the fluorescamine did not label the same subpopulation of cells although the lymphocytes could then be examined in the cell sorter. A larger number of cells (38.8%) formed the brighter cluster but did not behave as B cell when separated on nylon-wool columns. Improvement in discrimination of the cell populations was obtained with FITC-hexadecylamine (C16). This probe detected 38% of cells in the smallest cluster, 44% of cells in the intermediate cluster and 19% of cells in the brightest cluster. The proportion of cells in each cluster appeared to parallel closely the "null", erythrocyte (E) rosette-forming T cells and the B cells detected by conventional markers for blood lymphocytes. Other fluorescent probes, formed from FITC and other amines and amino acids, labelled lymphocyte membranes. Probes with a terminal charge labelled the small cluster particularly well, whereas those that were terminally non-polar labelled the larger cells brigthly, but not to the same intensity as the charged probes in the small cells.
绵羊血淋巴细胞用荧光探针标记，然后在荧光显微镜下和通过荧光激活细胞分选仪进行检测。一种由荧光胺与己胺偶联而成的新型探针，通过荧光显微镜计数，检测到22.9%的细胞，这些细胞显然属于B细胞系。用异硫氰酸荧光素（FITC）替代荧光胺后，虽然淋巴细胞随后可在细胞分选仪中进行检测，但并未标记相同的细胞亚群。大量细胞（38.8%）形成更亮的簇，但在尼龙毛柱上分离时其行为不像B细胞。使用FITC-十六胺（C16）可提高对细胞群体的区分度。该探针在最小的簇中检测到38%的细胞，在中间簇中检测到44%的细胞，在最亮的簇中检测到19%的细胞。每个簇中的细胞比例似乎与通过常规血液淋巴细胞标志物检测到的“裸”细胞、红细胞（E）花环形成性T细胞和B细胞密切平行。由FITC与其他胺类和氨基酸形成的其他荧光探针标记淋巴细胞膜。带有末端电荷的探针能很好地标记小簇细胞，而末端为非极性的探针能明亮地标记较大细胞，但亮度不如小细胞中带电荷的探针。
