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The XE-2100

The XE-2100. WBC and Differential measurement. P rinciples of cell detection. Wbc and differential counts are determined using : Flow Cytometry Method Semiconductor Laser. Blood cell information is obtained using: Forward light scatter Lateral light scatter Lateral fluorescent light.

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The XE-2100

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  1. The XE-2100 WBC and Differential measurement

  2. Principles of cell detection • Wbc and differential counts are determined using: • Flow Cytometry Method • Semiconductor Laser. • Blood cell information is obtained using: • Forward light scatter • Lateral light scatter • Lateral fluorescent light

  3. Optics

  4. Specific Measurements

  5. • Staining Reagent: – Polymethine Dye (Group of staining reagents): X = O, S, Se, N-alkyl or C(Ch ) 3 n R = alkyl (1-6C) ; R 1,2 3,4 = Alkyl, Methoxy or NO n = 0-1 The Stain

  6. The Wavelength • The new stain opens up the possibility of using a different laser wavelength on a compact laser system with little need of increases in power •  = 633nm • Red laser beam

  7. Methodologies

  8. WBC/BASO Channel • Blood sample is aspirated, diluted to a ratio of 1:50 with STOMATOLYSER-FB. • Diluted sample is sent to WBC/BASO detection chamber. • In this channel, forward and side scatter signals are used to derive the WBC and BASO count.

  9. Measurements • Forward Scatter - Measurement of size • Side Scatter - Measurement of internal structure i.e size of nucleus

  10. BASO Other WBC RBC Lysing Surfactant The WBC/BASO Channel

  11. WBC/BASO Scattergram

  12. WBC Classification • Blood sample is aspirated and diluted with: STOMATOLYSER-4DL and STOMATOLYSER -4DS. • Diluted sample is sent to the detector chamber • Cells are classified according to side scatter and fluorescence signals in the Diff channel.

  13. Measurements • Side scatter - measurement of internal structure. • Fluorescence - measurement of nucleic acid content.

  14. High Fluorescence Intensity Abnormal Lymph/ Lymphoblast High Fluorescence Intensity Low Fluorescence Intensity 1.Lysing 2.Staining The DIFF Channel Blast, I.G. Normal Cell STROM-4DL STROM-4DS Lysing surfactant Polymethine Dye +Organic Acid

  15. Diff Scattergram

  16. Neutrophil Count • This is a calculated parameter • # Neut = Neut/Baso count(diff channel)-Baso count(WBC/Baso channel)

  17. IMI Channel • discriminates between immature and mature white blood cells. • Detection is performed using RF/DC detection method.

  18. Methodology • Blood is aspirated and diluted to a ratio of 1:250 with STOMATOLYSER-IM. • Diluted sample is sent to the IMI detector for analysis. • A two-dimensional distribution of cell size and cell structure is drawn according to RF/DC detection method.

  19. Measurements • Direct current - measurement of cell volume • Radio frequency - measurement of internal cell structure.

  20. Platelet Clumps IMI Scattergram

  21. ACAS & Advanced Discrimination

  22. Adaptive Cluster Analysis System • Cells are classified by the process of cluster analysis which is based on learning algorithms. (An astronomical technique!) • This allows optimal adaptation to biological differences between samples including abnormal cells

  23. NLME:WBC except BASO Bas/NLME Gst/NLME Advanced Discrimination DIFF-CH WBC/BASO-CH 150+-1 Ly/Ne2 Ly/Mo2 150+-1 Mo/Eo Ly/Mo1 Mo/NE Ne/Eo Ly/Ne1 Ly/Ba Ly/Gst Ba/Gst Ne/Gst

  24. Extended ACAS Principle • High adaptation to individual changes • Moving clusters • Moving discriminators • Enhanced separation in abnormal cases • Low “Vote Out” rate

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