Hi,
What do you want to study?
1. Because if you want just characterized by specific lymphocytes populations, flow cytometry is a metodología.
2. But if you want to isolate specific lymphocytes subpopulatios both metodologies can help you.
1. With MACS you shlould to buy the magnetic beads conjugated to specific antibody with recognized an exclusive molecule in your interest lymphocytes population (positive selection) or thing in a negative selection.
2. Respect flow cytometry, you can label your lymphocytes subpopulation and then to separe by cell sorting.
Bye
thanks a lots , I want to know more about MACS
I didn't know what is the difference between positive selection or negative selection of lymphocyte population ?
Hi dear,
MACS methodology is based in specific antibodies conjugaste to magnetice bead, for example if you want to purified CD8 T cells you should to buy antibody anti-CD8 (conjugaste to magnetice beads).If you decide to use a positive selection (+) it means that your CD8 T cells labeled with specific antibodies are going to be retained in the column holder by the magento. You should to wash and at the end you elute CD8 T cells.
In you have to purified your CD8 cells by negative seleccion it means that your cells are not labeled and retained in the column. MACS have a antibodies mix for to remover mononuclear cells but not CD8 T cells.
You can go into MACS web and to read and undersand this protocol
Greetings
thank you for your help and rapid response.
usually when I prepared peripheral blood mononuclear cell and then I add antibody till analysis in flow cytometer I didn't understand the result so I try to understand the figures . in my lab all experiments on CD4 ,CD8 and MHC class 2 by using different types of antibodies such as PE,FITC,APC.
if you can help me in this point.
Hi Amany,
In flow citometry (FACS) the interesting cell are marked with specific Abs against specific cell populations markers as CD3, CD4, CD8, etc.
This Abs should be conjugated with a fluorocrome (PE, FITC, APC, PerCp,etc). This fluorocromes are excite by láser and the emition is in different spectrum (nm). During the analysis you observe dots and each one corresponde to a cell. In your Dot graphic in your cells are labeled with Ab-FITC these will be in an sito corresponde to FITC.
At the end, my suggestion is to review flowcytometry in a immunology techniques journal or book.
BYE
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