After two years of Tdt staining I still believe in the original procedure suggested by Zbigniew Darzynkiewicz. In our hands a solution 4% paraformaldeide or any other fixative, followed by 70% cold ethanol works almost all the time with every sample. Ethanol gives you also the opportunity to store your sample for a long time. l used the same sample fixed and stored in ethanol (-20) for almost two years as control reaction with good reproducible results. We also used this procedure for two color analysis (MoAb PE + Tdt Fitc),( we hope the result will be soon published in Blood) good luck Ugo Consoli Istituto di Ematologia Universita' di Catania, Italy Fax: +39-95-7311510 Tel: +39-95-7435917 Dear collegues > >I wonder how many laboratories do the nuclear TdT stain >by flow cytometric method after membrane permeabilization >not by cytospin followed by fluorscence microscopic reading? > >In case of flow cytometric method, which protocol for membrane >permeabilization you use? >Is the permeabilization reagent Triton-X, saponin, FACS Lysing Solution, >FACS Permeabilization Solution, Fix&Perm, or something special? > >And, have you ever tried to compare the results between >flow cytometric method and cytospin method? > >Any comments will be appreciated. >Thanks in advance. > >--------------------------------- >Chang-Seok Ki, M.D. >Dept. of Clinical Pathology >Samsung Medical Center >Seoul, Korea >Tel. 82-2-3410-2708 >Fax. 82-2-3410-2719 >E-mail. kcdol@samsung.co.kr >---------------------------------
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