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1. CD45 1 K3 EDTA 2 12X 75mm BD Falcon 352052 352054 352058 TruCOUNT 3 TruCOUNT BD 340334 IgG1 IlgG1 CD45 BD 349202. CD4 CD8 CD3 CD3 CD4 CD45 CD3 CD8 CD45 CD3 CD16 CD56 CD45 CD3 CD19 CD45 21300AMY00266000 21300AMY00263000 21300AMY00265000 21300AMY00262000 21300AMY00264000 T K 06 20 9 1 CD4 CD8 CD3 BD Tritest CD4 CD8 CD3 T T CD3 CD4 CD45 BD Tritest CD3 CD4 CD45 T T CD3 CD8 CD45 BD Tritest CD3 CD8 CD45 T NK CD3 CD16 CD56 CD45 BD Tritest CD3 CD16 CD56 CD45 T B CD3 CD19 CD45 BD Tritest CD3 CD19 CD45 1 2
3. PerCP CD45 CD4 T B NK CD45 TruCOUNT 1 ACD 20 25C CD4 CD8 CD3 48
4. NK B 488nm FITC 530nm PE 585nm PerCP 670nm PerCP CD4 CD8 CD3 PerCP CD3 CD3 T A
5. S 728 V 1 121C 20 1 000ppm 1 2 0 1 3 2 87 CD4 CD8 CD3
6. 4 CD3 PerCP RE FL3 488nm 650 700nm vs SSC 1 CD3 FITC FL1 FITC 488nm 15 545nm vs PE FL2 PE 488nm 562 607nm TruCOUNT BD MultSET 2 3 CD4S PerCP FL3 488nm gt 65 0nm
7. lt 1 HIV HBV HCV 2 FACS Lysing Solution 10 3 1 2 8 20 25C 3 4
8. CD3 CD8 CD45 Q1 Q4 Q1 Q2 Q3 Q4 CD3 CD8 CD3 CD8 CD3 CD8 NK TT CD3 CD8 TAME CD3 CD16 CD56 CD45 Q1 Q4 Q1 Q2 Q4 CD3 CD16 CD56 CD3 CD16 CD56 CD3 CD16 CD56 NK TT TT CD3 CD19 CD45 Q1 Q4 Q1 Q3 Q4 CD3 CD19 CD3 CD19 B CD3 CD19 95 Range T 31 60 13 41 55 84 T T 96 B 6 25 S 27 410 1590 NK 96 T L 190 1140 690 2540 90 660 FIL y Y AOS T L T L 90 590 IT ML B L NK L 18 65
9. ZM K3 EDTA 20 25 lt gt 1 TruCOUNT 1 2 20 25C 20gL TruCOUNT 3 50 L TruCOUNT 4 Vortex 20 25C 15 NL 5 10 FACS Lysing Solution 4504L 4 MX Fo 10 FACS Lysing Solution 20 25C
10. 24 6 24 CD3 CD4 CD45 CD3 CD8 CD45 72 24 48 24 6 6 24 6 24 6 CD3 CD16 CD56 CD45 CD3 CD19 CD45 MIE AE 24 6 6 1 2 3
11. 3 4 S FITC CD4 CD4 CD8 CD3 PE CD8 FITC CD3 R247 4A CD3 CD4 CD45 PE CD4 FA FAT CD3 CD8 CD45 FITC CD3
12. CD3 CD16 CD56 CD45 NK 0 92 wh R WB T NK zL T gL 0 23 0 21 0 94 07 035 e 52 0 96 26 94 109 0 94 100 3790 TruCOUNT CV NK GAEL 4 1 N 10 1360 T L CD4 CD3 CD19 CD45 B T N Um R SE pH 167 0 94 1 6 0 94 0 44 3 7 0 96 24 94 B L 167 0 91 166 0 97 0 95 0 1370 24 T L 118 0 95 130 3710 TruCOUNT N 10 1197 B L T L CD4 WBC 2 5 x 10 31 0 10 L 2 0x 10 16 7 x 105808 LL
13. v 6 Vortex 20 25C 15 20 25C 1 2 CaliBRITE 3 CaliBRITE 3 CD4 CD8 CD3 3 FSC SSC
14. CD3 CD4 CD45 CD3 CD8 CD45 CD3 CD16 CD56 CD45 CD3 CD19 CD45 SOFA F 1 Schmidt RE Monoclonal antibodies for diagnosis of immunodeficiencies Blut 1989 59 200 206 2 Nicholson JKA Use of flow cytometry in the evaluation and diagnosis of primary and secondary immunodeficiency diseases Arch Pathol Lab Med 1989 113 598 605 3 Foucar K Goeken JA Clinical application of immunologic techniques to the diagnosis of lymphoproliferative and immunodeficiency disorders Lab Med 1982 13 403 413 4 Cohen SB Weetman AP Activated interstitial and intraepithelial thyroid lymphocytes in autoimmune thyroid disease Acta Endocrinol 1988 119 161 166 5 Smolen JS Chused TM Leiserson WM Reeves JP Alling D Steinberg AD Heterogeneity of immunoregulatory Tcell subsets in systemic lupus erythematosus correlation with clinical features Am J Med 1982 72 783 790 6 Giorgi JV Hultin LE Lymphocyte subset alterations and immunophenotyping by flow cytometry in HIV disease Clin Immunol Newslett 1990 10 55 61 7 Landay A Ohlsson Wilhelm B Giorgi JV Application of flow cytometry to the study of HIV infection AIDS 1990 4 479 497 8 Antel J Bania M Noronha A Neely S Defective suppressor cell function mediated by T8 cell lines from patients with progre
15. L T L CD4 CD3 CD4 CD45 ERS N T T 96 168 T gL T gL TruCOUNT Peas Tann ove o 2034 2716 T L 1897 CD4 CD3 CD8 CD45 ERS N T 168 0 98 T 96 168 L T 194 0 22 1 06 10 0 98 70 1980 T gL 11 0 99 120 2860 TruCOUNT N 10 gL TT CAE uL CD4
16. SSC Ca EL1 FITC 488nm 30 15nm vs FL2 PE 488nm 58 21nm FL1 FL2 8 4 CD45 PerCP FL3 vs SSC 3 5 ETITC FL1 vs PE FL BD MultiSET 4 7 TruCOUNT BD MultiSET MLIT CD8 P
17. 1 2 3 8 4 5 CD4 T 6 CD8 NK CD4 CD8 CD3 a Ton sm Ei T 143 1 00 BEARER TruCOUNT N 10 2048 1311
18. PE CD8 CD3 CD16 CD56 CD45 FITC CD3 PE CD16 PE CD56 FITC CD3 PE CD19 CD3 CD19 CD45 CD4 CD8 CD3 T T CD3 CD4 CD45 T TT CD3 CD8 CD45 CD3 CD16 CD56 CD45 CD3 CD19 CD45 TT
19. How to Define and Determine Reference Intervals in the Clinical Laboratory Approved Guideline Villanova PA National Committee for Clinical Laboratory Standards 1992 NCCLS document C28 A Lanier LL Le AM Phillips JH Warner NL Babcock GF Subpopulations of human natural killer cells defined by expression of the Leu 7 HNK 1 and Leu 11 NK 15 antigens J Immunol 1983 131 1789 1796 Free Dial 0120 8555 90 FAX 024 593 5761 T 960 2152 1 Free Dial 0120 8555 90 P 64 030 7
20. University Press 1989 517 542 Ritz J Trinchieri G Lanier LL NK cell antigens section report In Schlossman SF Boumsell L Gilks W et al eds Leucocyte Typing V White Cell Differentiation Antigens New York NY Oxford University Press 1995 2 1367 1372 Nadler LM B Cell Leukemia Panel Workshop summary and comments In Reinherz EL Haynes BF Nadler LM Bernstein ID eds Leukocyte Typing II Human B Lymphocytes New York NY Springer Verlag 1986 2 3 43 D rken B M ller P Pezzutto A Schwartz Albiez R Moldenhauer G B cell antigens CD19 In Knapp W D rken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford University Press 1989 34 36 Berti E Parravicini C Cattoretti G Delia D de Braud F Cusini M Immunohistochemical reactivity of anti B cell monoclonal antibodies in thymus lymph node and normal skin In Reinherz EL Haynes BF Nadler LM Bernstein ID eds Leukocyte Typing Il Human B Lymphocytes New York NY Springer Verlag 1986 2 313 318 Centers for Disease Control Update universal precautions for prevention of transmission of human immunodeficiency virus hepatitis B virus and other bloodborne pathogens in health care settings MMWR 1988 37 377 388 Protection of Laboratory Workers from Infectious Disease Transmitted by Blood Body Fluids and Tissue Tentative Guideline Villanova PA National Committee for Clinical Laboratory Standards 1991 NCCLS do
21. 2 BD Multi Check control BD 340911 340912 340913 6 FACS Lysing Solution 10X BD 349202 20 25 20 25C 10 20 25C 20 25C 1 7 BD FACSFlow 342003 8 BD CaliBRITE3 340486 9 LA 10 Vortex 11 12 13 488nm FACS
22. E CD3 PerCP CD4 FITC 1 CD4 CD8 CD3 2 CD3 FL3 vs SSCT CD4 CD8 3 SSC CD4 PE 107 of CD3 FITC 4 CD45 FL1 vs FL2 CD3 CD4CD45 ne CD45 PerCP 3 FL3 vs SSC CD45 CDS PE CD16 CDS6 PE CD3 FITC BIS CD45 FL1 vs FL2 CD3 CD8 CD45 CD3 FITC 6 CD45 FL1 vs FL2 CD3 CD16 CD56 CD45 a 2 e O Q1 Q4 gt CD3 FITC Q1 Q2 7 CD45 o qq FL1 vs FI2 CD3 CD19 CD45 Q3 Q4 FL1 8 FL1 vs FL2 CD4 CD8 CD3 Q1 Q4 Q1 Q4 CD3 CD8 CD3 CD4 CD3 CD4 CD45 Q1 Q4 Q2 Q3 Q4 CD3 CD4 CD3 CD4 CD3 CD4
23. cument M29 T2 Nicholson JK Browning SW Orloff SL McDougal JS Inactivation of HIV infected H9 cells in whole blood preparations by lysing fixing reagents used in flow cytometry J Immunol Methods 1993 160 215 218 Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture Approved Standard Villanova PA National Committee for Clinical Laboratory Standards 1991 NCCLS document H3 A3 Clinical Applications of Flow Cytometry Quality Assurance and Immunophenotyping of Peripheral Blood Lymphocytes Tentative Guideline Villanova PA National Committee for Clinical Laboratory Standards 1992 NCCLS document H42 T Giorgi JV Lymphocyte subset measurements significance in clinical medicine In Rose NR Friedman H Fahey JL eds Manual of Clinical Laboratory Immunology 3rd ed Washington DC American Society for Microbiology 1986 236 246 Jackson AL Warner NL Preparation staining and analysis by flow cytometry of peripheral blood leukocytes In Rose NR Friedman H Fahey JL eds Manual of Clinical Laboratory Immunology 3rd ed Washington DC American Society for Microbiology 1986 226 235 Prince HE Hirji K Waldbeser LS Plaeger Marshall S Kleinman S Lanier LL Influence of racial background on the distribution of T cell subsets and Leu 11 positive lymphocytes in healthy blood donors Diagn Immunol 1985 3 1 33 37 Angadi CV Lack of Leu 3a epitope on T helper CD4 lymphocytes J Clin Lab Anal 1990 4 193 195
24. en using the whole blood lysis procedure and flow cytometry Cytometry 1996 26 16 21 Evans RL Wall DW Platsoucas CD et al Thymusdependent membrane antigens in man inhibition of cellmediated lympholysis by monoclonal antibodies to the TH2 antigen Proc Natl Acad Sci USA 1981 78 544 548 Bernard A Boumsell L Hill C Joint report of the first international workshop on human leucocyte differentiation antigens by the investigators of the participating laboratories T2 protocol In Bernard A Boumsell L Dausset J Milstein C Schlossman SF eds Leukocyte Typing New York NY Springer Verlag 1984 25 60 18 Wood GS Warner NL Warnke RA Anti Leu 3 T4 antibodies react with cells of monocyte macrophage and Langerhans lineage J Immunol 1983 131 212 216 Haynes BF Summary of T cell studies performed during the Second International Workshop and Conference on Human Leukocyte Differentiation Antigens In Reinherz EL Haynes BF Nadler LM Bernstein ID eds Leukocyte Typing II Human T Lymphocytes New York NY Springer Verlag 1986 3 30 Kan EAR Wang CY Wang LC Evans RL Noncovalently bonded subunits of 22 and 28 kd are rapidly internalized by T cells reacted with Anti Leu 4 antibody J Immunol 1983 131 536 539 Knowles RW Immunochemical analysis of the T cell specific antigens In Reinherz EL Haynes BF Nadler LM Bernstein ID eds Leukocyte Typing II Human T Lymphocytes New York NY Springer Verlag 1986 1 259 288 Organ
25. izing Committee of the Fourth International Workshop on Human Leucocyte Differentiation Antigens Appendix A CD guide In Knapp W D rken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford University Press Inc 1989 1074 1093 Dalgleish AG Beverley PCL Clapham PR Crawford DH Greaves MF Weiss RA The CD4 T4 antigen is an essential component of the receptor for the AIDS retrovirus Nature 1984 312 763 767 Maddon PJ Dalgleish AG McDougal JS Clapham PR Weiss RA Axel R The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain Cell 1986 47 333 348 Rudd CE Burgess KE Barber EK Schlossman SF Monoclonal antibodies to the CD4 and CDS antigens precipitate variable amounts of CD4 CD8 associated p56lck activity In Knapp W Dorken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford University Press 1989 326 327 Schwinzer R Cluster report CD45 CD45R In Knapp W Dorken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford University Press 1989 628 634 Moebius U Cluster report CD8 In Knapp W Dorken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford University Press 1989 342 343 Gallagher PF Fazekas de St Groth B Miller JFAP CD4 and CD8 molecules can physically associate with the
26. lynas DP et al Identification of a putative second T cell receptor Nature 1986 322 145 149 Clevers H Alarc n B Wileman T Terhorst C The T cell receptor CD3 complex a dynamic protein ensemble Annu Rev Immunol 1988 6 629 662 Fitzgerald Bocarlsy P Herberman R Hercend T et al A definition of natural killer cells In Ades E Lopez C eds Natural Killer Cells and Host Defense Basel Karger 1989 1 Lanier LL Le AM Civin CI Loken MR Phillips JH The relationship of CD16 Leu 11 and Leu 19 NKH 1 antigen expression on human peripheral blood NK cells and cytotoxic T lymphocytes J Immunol 1986 136 4480 4486 Perussia B Acuto O Terhorst C et al Human natural killer cells analyzed by B73 1 a monoclonal antibody blocking Fc receptor functions II studies of B73 1 antibody antigen 40 41 42 43 44 45 46 47 48 49 30 51 52 53 34 RN 5S6 interaction on the lymphocyte membrane J Immunol 1983 130 2142 2148 Perussia B Starr S Abraham S Fanning V Trinchieri G Human natural killer cells analyzed by B73 1 a monoclonal antibody blocking Fc receptor functions I characterization of the lymphocyte subset reactive with B73 1 J Immunol 1983 130 2133 2141 Schmidt RE Non lineage natural killer section report new and previously defined clusters In Knapp W D rken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford
27. same T cell receptor Proc Natl Acad Sci USA 1989 86 10044 10048 Anderson P Blue M L Morimoto C Schlossman SF Cross linking of T3 CD3 with T4 CD4 enhances the proliferation of resting T lymphocytes J Immunol 1987 139 678 682 Eichmann K J nsson J I Falk I Emmrich F Effective activation of resting mouse T lymphocytes by cross linking submitogenic concentrations of the T cell antigen receptor with either Lyt 2 or L3T4 Eur J Immunol 1987 17 643 650 Wood GS Warner NL Warnke RA Anti Leu 3 T4 antibodies react with cells of monocyte macrophage and Langerhans lineage J Immunol 1983 131 212 216 Moebius U Cluster report CD8 In Knapp W Dorken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford University Press 1989 342 343 Schwinzer R Cluster report CD45 CD45R In Knapp W Dorken B Gilks WR et al eds Leucocyte Typing IV White Cell Differentiation Antigens New York NY Oxford University Press 1989 628 634 Cobbold SP Hale G Waldmann H Non lineage LFA 1 family and leucocyte common antigens new and previously defined clusters In McMichael AJ ed Leucocyte Typing III White Cell Differentiation Antigens New York NY Oxford University Press 1987 788 803 van Dongen JJM Krissansen GW Wolvers Tettero ILM et al Cytoplasmic expression of the CD3 antigen as a diagnostic marker for immature T cell malignancies Blood 1988 71 603 612 Brenner MB McClean J Dia
28. ssive multiple sclerosis J Immunol 1986 137 3436 3439 9 Gratama JW Naipal A Oljans P et al T lymphocyte repopulation and differentiation after bone marrow transplantation early shifts in the ratio between T4 and T8 T lymphocytes 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 23 26 21 28 29 30 31 32 33 34 35 36 37 38 39 correlate with the occurrence of acute graft versus host disease Blood 1984 63 1416 1423 Bishop GA Hall BM Duggin GG Horvath JS Sheil AGR Tiller DJ Immunopathology of renal allograft rejection analyzed with monoclonal antibodies to mononuclear cell markers Kidney Internat 1986 29 708 717 Wolde Mariam W Peter JB Recent diagnostic advances in cellular immunology Diag Med 1984 7 25 32 Centers for Disease Control 1997 Revised guidelines for performing CD4 T cell determinations in persons with human immunodeficiency virus HIV MMWR 1997 46 No RR 2 1 29 Nicholson JKA Jones BM Hubbard M CD4 Tlymphocyte determinations on whole blood specimens using a single tube three color assay Cytometry 1993 14 685 689 Nicholson J Kidd P Mandy F Livnat D Kagan J Threecolor supplement to the NIAID DAIDS guideline for flow cytometric immunophenotyping Cytometry 1996 26 227 230 Nicholson JKA Hubbard M Jones BM Use of CD45 fluorescence and side scatter characteristics for gating lymphocytes wh
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