On the active E rosette test in the optimization of experimental conditions.docVIP

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On the active E rosette test in the optimization of experimental conditions.doc

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On the active E rosette test in the optimization of experimental conditions

On the active E rosette test in the optimization of experimental conditions Abstract: Active E rosette test on how to optimize the experimental materials, reagents, operating and other experimental conditions, combined with practical work experience are described, while the main effect of experimental factors were discussed to improve the success rate of the experiment . Keywords: E rosette test SRBC lymphocytes, E rosette test is the number of in vitro detection of T lymphocytes and immune function in a classical way, different reaction conditions to form a different knot. T lymphocytes and a lower proportion of sheep red blood cells (SRBC by at 37 ℃, After low-speed centrifugation can develop without the formation of cold knot called the active E rosette (Ea-RFC, E rosette activity represents a high affinity with the SRBC activity of T cell subsets [1], able to respond cellular level, often with the same clinical manifestations. Active E rosette test although no special equipment, but the impact of many factors, reproducibility and stability difficult to control, control of experimental conditions to test the key to success, and thus the author of the Ea-RFC pilot The main experimental conditions were summarized and optimized to improve the success rate of the experiment. 1 Isolation of peripheral blood mononuclear cells 1.1 The collection of blood samples to be collected blood samples seized the traditional method is to use disposable syringe after the injection of test tubes, and vacuum blood is the blood of the new generation product to replace syringes, blood sampling may be the use of disposable lancet blood collected in heparin anticoagulant built agent or lymphocyte separation medium of vacuum blood, so to avoid the use of traditional syringe suction damage on the role of blood cells, and can shorten the time of blood collection. picked the blood For placement, should be set to 4 ~ 8 ℃ low temperature and not more than 6h, and the s