Counting thymocyte population - (Jun/21/2008 )
Since my lab has no flow cytometer and other sophisticated equipments to instantly count cells, I have no idea how to count thymocyte population in valid way. I never did this before. Then I think this way: I use hemacytometer with this simple protocol: Thymus capsule is torn to release its cells --> then pour them into PBS (pH 7.2) at certain volume we decide (depends on the mass of the thymus) --> I centrifuge the solution to separate thymocyte and other tissues --> the precipitate then is rinsed with PBS once more --> follow the protocol to count RBC/WBC.
What do you think? I really need suggestion/advice before I do the real research next month.
Thank you in advance 
U mean mouse thymocytes, do U?
Make thymocyte cell suspension and proceed like U count WBC with appropriate dilution with Trypan blue. They are merely lymphocytes - round and bright.
If mouse thymocytes, make single cell suspension with the wrong end of a syringe by crushing them in a culture plate/petri dish with little PBS (or ACK if U have too much red cells), filter them through fine mesh and wash them with PBS. I make cell suspension of 4-5ml for one thymus and dilute 40X to count them in hemocytometer like U count WBC.
excuse me for I have no idea.because I did not use this methot.
Then celebrate
No need to reply 
Then celebrate
No need to reply Cheers! Meet U in lunch time then
Make thymocyte cell suspension and proceed like U count WBC with appropriate dilution with Trypan blue. They are merely lymphocytes - round and bright.
If mouse thymocytes, make single cell suspension with the wrong end of a syringe by crushing them in a culture plate/petri dish with little PBS (or ACK if U have too much red cells), filter them through fine mesh and wash them with PBS. I make cell suspension of 4-5ml for one thymus and dilute 40X to count them in hemocytometer like U count WBC.
Thanks for the reply, B'Boy. I really need that
I'm happy because my method is not wrong at all 
Yes, I meant thymocytes from mouse and rats.More questions,
1. About trypan blue, do we have to apply it in certain amount or not? If I am not mistaken, it is for background colour to make thymocytes is clearly seen (due to the contrast), right? Or else?
2. Can I adopt this method to perform lymphocytes count from mouse/rat spleen too? Or I need special method, which is different, since spleen contains many kinds of cells, more complicated that thymus.
Thanks for the help
@celebrate & cellcounter: Meet u in lunch time too
O, we missed this posting. 
Sorry. We are late.
Yes, lymphocytes from anywhere I guess.
Spleen contains many kinds of cells??? What kinds of cells are U seeing?