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????? ??????? ????????????? ????????? ??? ?? ??????????? ?? ?????? ?????????? ?? ????? ?????????? ?????????? ??? ??? ???????? ???????? ????. ???????????? ??????????? ???? ????? ??? ???????????, ???? ??????, ???? ????????,???? ?????????? ????????, ??????????? ? ?? ??????. ?? ?????????? ????? ?????????????? ????? ???????? ?????. ??? ????????? ???? ???????? ????????????????????????? ??? ?????????. ? ?????? ????? ????? ??? ???? ?????????? ????????????? ??????? ??? ???? ?? ???? ????????????? ???????? ??? ????? ?????? ??? ???????????, ?? ?- ???????????? ? ?? ?????????? ??? ??????????? ??? ?? ?-????????????.???????????, ? ?????? ??? ? ????? ?????? ???????????? ????????????? ?????? ??????’ ???? ????????????? ?? ???????????? ??? ?????????? ???? ?????? ????? ????????. ?? ????????? ???????????? ??? ????????? ??? ????? ?????? ??????????????????? ??? ??????? ???? ??? ????????????? ??? ??? ??????????????????. ? ?????? ???????? ????? ???? ????????????? ????????? ?? ??????????????? ????????????? ?? ????? ???????? ??????????.

?? ???? ????????? ????? ?????? ????????????? ??? ????????, ????????? ????????????? ??? ???????????. ?????? ???????? ??? ????????, ????? ??? ??????????? ???? ???????? ??? ?-????????????? ??? ??????? ?? ???? ??? ????? ??????????? ??? ??????????? ?? ?????????. ? ????????????? ????(Flow Cytometry, FC) ????? ??? ??????? ????????????????? ?????????? ??????????? ????????? ?? ??????? ??????????? ?????????? (????????, ???????,????????????). ?? ????????????? ??? FC ????????? ?????? ??? ?????????? ????????? ?? ?????? ????????, ????? ??? ?? ????? ????????, ?????????? ?????????????? ? ??? ?????? ?????????????? ??? ????????.

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?? ??????????????? ???? ????????????? ?????? ????? ??? ????? laser  ? ??????????????? ????? ??? ??????? ????? ????? ???????????? ?????????? ?? ??? ???? ??????. ? ??????????? ??? ???? ??????? ??????????? ?????????? ??? ????????????????????? ??? ???????????? ???? ??????????, ???????????? ??????????? ??? ????????? ???????, ??? ????????, ??? ????????????? ?????, ??? ???????????????????? ? ????????????? ?????????, ?? ??????????? DNA ??? ??? ??????????????? ???????????.??????????, ?? ???????????? ? ?? ??????? ??? ??????? ??? ????????, ?????????????? ?????? ??? ??????? ???? ??? ?????? ??? ?? ??????, ???????????? ???????? ?? ??? ????? ?????? ??? ??? ??????? ??? ??? ???? ?????. ?? ??????????????? ??? ?? ????? ??????? ??????????? ??????? ?? ?? ??????? ???? ??? ???????????? ?????????????. ?????????, ?? ??????? ??? ????? ??????????? ???????????? ???????o? ????????? ??? ?? ??????????? ???? ?????? ????? ???????. ????? ??? ?????????? ??? ?????????? ????????? ?? ????????????? ??????????(filters) ??? ?????????? ???? ??? ????????????.????????? ????????????? ??????? ????? ?? ???????????? ??? ???? ????????? ????????? ????? ????? ??????? ?????? ????????????? ?? ?????????? ?? ????????? ? ??? ??????????????. ??? ?? ????? ???? ?? ?????????????? «???????» ??? ?????????????? ????? ?????????? ??? ?? ???????? ??????????? ??? ??????? ? ??? ? ??????????? ?? ?????????? ??? ????????? ?????????.

?? ??????? ??????????? ?????????????? ?????????? ???????? ????? ??????? ??? ?????????? ??? ? ??? ????????: CD4 (??? ?????????? ?????? ?? ? ????????? ???????), CD8 (????????????? ?????? ?? ????????????? ? ???????) CD3 (??? ?????????? ?? ??? ??????? ??????? ? ?? ????? ??? ????????????? TCR) ??? CD19 ? CD45R (????????????? ???? ????????? ??? ?????? ? ????????). ??? ????????, ???? ?????????? ????????????? ??? ????????????, ?? ????????? ?? ?????????????? ?? ??????? ??????? ?????????? (flowjo).?????????? ?????????????? ?????? ?? ?? ??????? ??? ?????  ??????????? ?????? ??? ?? ???????????? ?? ???? ??????? ????????. ???????,????????? ???????????? ??? ?????????.

??????????????? ?? ??????? ??? ?? ??????? ???? ?????????? ?? PBS ???????????? ?? 5% FCS ??? 0,01% ?????? ??? ???????.?????????, ?? ?????????? ??????? ??? ??????????? ??? ?? ?????? ?? ??????? ???????? ?????????????? ?????????? ??? 5 ?????. ??? ????????, ???????????? 2 ml FACSbuffer ??? ???????????????.  ??????,?????????? ?? ??????? ??????? ???????????  ?? ?? ????? ?????? neubauer ???????????? ?rypan blue. ?????????????? ??? ??????? ??????????? ??? ???????? ???????????? anti-CD3, antiCD4, anti-CD8 ??? anti-CD19.

??????????? ?? ?????????????????? ???? ??????? ??? ?????????, ?? ???????? ???????????????  ??? ??????????  ??? 30 ????? ??? ???? ? ??? ??????.??????????? ?? ????????  ?? FACS buffer??? ?????.  ??????????????? ?? ??????? ????????? PFA 1% ??? 15 ?????.

  ?? ????????????????? ?? FACS buffer ??? ?? ??????? ??????????????? ?? FACS buffer.  ?? ‘??????? ???????’ ???????????? ?? ??????tubes ??? ???????????? ??? ????? ?? ????????????. ?? ???????  ????? ?????? ??? ??????? ??????????? ??????????????.