‘voutoonis 1 (ght Seater FIGURE 41-6 Discriminant function 1 (OF) scatterplet, Coulter ‘STKS. DF1 scatterplot graphs volume vs light scatter and re- vals the locations of four leukocyte populations. The basophil [population is located benina the iympnocytes. (Courtesy ot Coulter Electronics, nc., Hialeah, FL) Web Figure 41-1 DF2 scatterplot, Coulter STKS, DF2 scatterplot graphs volume vs. conductivity and reveals the locations of three leukocyte populations. The granulocyte population represents neutrophils, eosinophils, and basophils. (Courtesy of Coulter Electronics, Inc., Hialeah, Fla.) Copyright © 1995-2003 by Prentice-Hall, Inc, A Pearson Company Legal Notice Web Figure 41-2 DF3 scatterplot, Coulter STKS. This scatterplot also graphs volume vs conductivity, but the neutrophil and eosinophil populations are gated out of the granulocyte population to reveal the basophils. (Courtesy of Coulter Electronics, Inc., Hialeah, Fla.) Copyright © 1995-2003 by Prentice-Hall, Inc. A Pearson Company Legal NoticeCOULTER PRINCIPLE otras ectrode lood ce pass Conductive ing trough + i ‘erture “duen FIGURE 41-1 Coulter principle. Increased electrical resistance, ‘orimpedance, occurs when the poorly conductive blood cell asses through the aperture. Reprinted, with permission, from Pierre R. Seminar and Case Studies: The Automated Ditferen- tie Hialeah, FL: Coulter Electronics, inc.; 1985.) 25 26.27 28 29-90 Ot 42 33 36 95 36 3: femtoliters > | FIGURE 41-2 Histogram, or size distribution curve, allows the visualization ofthe biood cell population based on the relative Cell number and its volume (i 1). (Reprinted, with permission, ‘rom Significant Advances in Hematology. Hialeah, FL: Coulter Electronics, Inc; 1983.) | FIGURE 41-5 WBC histogram and count. In a normal patient, the Iymphooyte region represents lymphocytes, the mononuclear region represents monocytes, and the granulocyte region represents neutrophis, eosinophils, ané ba- ‘sophils, (Reprinted with permission, from Significant Advances in Hematology. Hialeah, FL: Coulter Electronics, inc. 1983.)IB FIGURE 41-3 RBC histogram and count, The shaded area represents those cells used in the ROW calculation. The ‘excluded celis may represent large platelets, platelet clumps, or electrical interference on the left and RBC dou- blets, REC triplets, RBC agglutinates, or aperture artifacts on the right. (Reprinted, with permission, from Significant “Advances in Hematology. Hialeah, FL: Coulter Electronics, Inc; 1983) RELATIVE NUMBER OF PLATELETS pores | st Ps [TAS eect oll 2.0m 20ym? PLATELET VOLUME 28 ym? FIGURE 41-4 Normal platelet histogram, Coulter Counter S Plus. The jagged line represents the raw data collected from 2-20 fL. The smooth line represents the extrapolated histogram from 0-70 fL. (Reprinted, ‘with permission, from Significant Advances in Hematology. Hialeah, FL: Coulter Electronics, Inc.; 1983)WBC sPeroc IE FIGURE 41-7 Location of abnocmal cell types on the scatter- plot, Coulter STKS. (Courtesy of Coutter Electronics, inc. Hialeah, FL) MMIETIN§ Coulter S-Plus WBC Histogram Region Flags [Region Flag Affected Poston Possible Abnormalities la Lymphocyte population Nuceated enthrooyte, large | ‘ees not boa at platlts, clumped pate bassline (-35 8) les, or naceluar parasites (9, materi td No valoy bacon Reactive ymohocytes, blast Iymphnocjta and mono- cals rear populations oon) 3 No valoy betwen Eoshophila, asopila, ‘mononuclearand Immature neatophis ‘reruooytepopuia tins (16011) a Granulocyte population Granuiooytoss oes not et to bssine (-450 ru Interrence detected in ‘mutiple postions (her impedance-oased netruments proving three-part laukcoyte dferentls ss fagging system simi to tho Cauiter S-Plus instrumenteT OFS ISTOGRAM ANALYSIS een "LAL | oo Date of Birth Sex User Field 1 Tocation User Field 2 Prysigian User Field 3 bate & Time 10/02/92 23:46 Conments IDE 1, 62069 wec 9.0 ACC # x Sequence # NE 64.7 5.8 19/02/92 m 4 ae 23:47:36 OuEti2 <8) 0:3, 5009341 BA 03 0:8 Norma WBC Pop Normal RBC Pop Normal PLT Pop FIGURE 41-8 Coulter STKS report from a normel individual. (Courtesy of Linda Nash, MT(ASCP), ‘Southwest Texas Methodist Hospital, San Antonio, TX)ight-ange scatter optics fa 64, Laser flow cylometty. The optical detection of forward- and right-angle light scatter using a laser light ‘cure is accomplished by using a sensor es the cells poss firough the beam under conditions of laminar flow. (Cour- 'tsy of Ortho Diagnostic Systems, Westwood, Mass., 1985) Sample and sheath stream Optical 2 sensor ch ° O Fhuorencent cl «droplet, 2 Computer 0 3 Mi fotanse 3 Fig. 165, Loser and cell sorting schematic. In a flow cytome- fry system, stained cells flow through o sample tube. As the «ells and o stream of saline solution leave the flow cham- ber, they move like a string of beads in the center of the sheath. The fluorescence of the cells is detected by a sen- sor. Cells can be appropriately chorged as they move through a charging collar or deflection plates. Sorting of the cells is accomplished by defleciing charged cells de- pending on the charge (either positive or negative).