Mege J L, Capo C, Benoliel A M, Foa C, Bongrand P
J Immunol Methods. 1985 Sep 3;82(1):3-15. doi: 10.1016/0022-1759(85)90219-4.
Cell deformability plays an important role in many immunological processes, such as phagocyte chemotaxis and endocytosis. The most widely used method of assay consists in aspirating cells into glass micropipettes and measuring the length of the protrusion induced by a given pressure, or the minimum pressure required to drive cells into the micropipette. This procedure requires specialized equipment and delicate manipulation. The present report describes a simpler procedure: cells are centrifuged in petri dishes floating on a water cushion, then fixed and coated with 0.8 micron diameter latex beads, which allows rapid and accurate determination of their height. This method is compared with the micropipette technique by studying lymphocyte and macrophage-like cell lines in physiological medium and in the presence of a divalent cation chelator or a microfilament inhibitor. In addition to simplicity, the main advantages of this technique are that (i) many cells may be examined within a reasonable period of time, which allows testing of heterogeneous cell populations, and (ii) unexpectedly, centrifugation was quite harmless under our experimental conditions, since it did not impair cell proliferative ability nor phagocytic ability. It is concluded that the method may be used in clinical laboratories to explore phagocyte dysfunctions, as well as in experimental studies.
细胞变形性在许多免疫过程中发挥着重要作用,如吞噬细胞趋化性和内吞作用。最常用的检测方法是将细胞吸入玻璃微量移液器中,并测量给定压力诱导的突出长度,或驱动细胞进入微量移液器所需的最小压力。该过程需要专门的设备和精细的操作。本报告描述了一种更简单的方法:将细胞在漂浮于水垫上的培养皿中离心,然后固定并用直径0.8微米的乳胶珠包被,这使得能够快速准确地测定其高度。通过在生理培养基中以及在存在二价阳离子螯合剂或微丝抑制剂的情况下研究淋巴细胞和巨噬细胞样细胞系,将该方法与微量移液器技术进行了比较。除了简单之外,该技术的主要优点是:(i)可以在合理的时间内检查许多细胞,这允许对异质细胞群体进行测试;(ii)出乎意料的是,在我们的实验条件下离心相当无害,因为它不会损害细胞增殖能力和吞噬能力。结论是该方法可用于临床实验室探索吞噬细胞功能障碍,也可用于实验研究。