Hello, Nadeem: Several years ago, while finishing my Ph.D dissertation at Harlow's Primate Laboratory (UW-Madison), I was involved in a similar project to look at lymphocytes and monocytes in CSF from disease-free Rhesus. We collected CSF drop by drop using cervical tapping without suction. It was about 50 ul per drop. We tried to collect 1.5 ml. We then use this sample for microscopic counting stained with acridine orange, mainly to estimate the total number of RBC and WBC, and for a 3-color flow cytometric analysis with a vital DNA stain for the nucleated cells. The inclusion of this vital DNA dye was quite helpful for us to discriminating nucleated cells from non-cellular debris, especially analyzing samples with limited number of cells. Besides, we created a tube insert by putting a 600-ul microfuge tube into a regular FACS tube to hold the sample for maximum acquistion. Please review the literature below and let me know if you have other questions. Leukocyte Trafficking in Free-Flowing Cerebrospinal Fluid of Normal Rhesus Macaques, Journal of Medical Primatology 25(1), 8-13, 1996, FY Hou, CL Coe, C Erickson Psychological Disturbance Differentially Alters CD4+ and CD8+ Leukocytes in the Blood and Intrathecal Compartments, Journal of Neuroimmunology 68, 13-18, 1996, FY Hou, CL Coe, C Erickson Fang-Yao Stephen Hou, Ph.D. Flow Cytometry Specialist UC-Irvine
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