Re: Nuclear TdT? Which method is better?

Ugo Consoli (uconsoli@mbox.vol.it)
Wed, 17 Sep 1997 20:52:32 +0200

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
>---------------------------------