최신 기종 사용 경험 (4)

Abbott : CELL-DYN 4000

원자력의학원
원자력병원
진단검사의학과
장윤환
CELL-DYN 4000 : 기능 및 구성
 Intended Use
‹ WBC parameters
WBC, NEU, %N, LYM, %L, MONO, %M, EOS, %E,
BASO, %B, WVF (WBC Viable Fraction)
‹ RBC parameters
RBC, HGB, HCT, MCV, MCH, MCHC, RDW, NRBC, NR/W,
RETC, %R, IRF (Immature Reticulocyte Fraction)
‹ Platelet parameters
PLT, MPV, PCT, PDW
 Specimen Processing Sequence

‹ Specimen Loading and Presentation

Autoloader mode
Open tube mode

‹ Specimen Identification and Test Selection

‹ Sample Dilution and Analysis

Sample Mixing
Sample Aspiration
CD4000 Flow Chart
CELL-DYN 4000
 Hardware and Sofeware
‹ System Hardware
Analyzer
Pneumatic Unit
Data Station
Printers

‹ System Software
Analyzer Operating Software
Data Station Operating Software
Screen Saver
 Reagents
‹ CELL-DYN 4000 Reagents
Diluent/Sheath Reagent
Hemoglobin Reagent
WBC Reagent - Parts A and B
Reticulocyte Reagent

‹ Controls, Calibrator, and Standard Reference

Particles
CELL-DYN 4000 : 측정 원리
 Principles of Operation
‹ Flow Cytometry
Flow Cytometry Measurement Optimization
Optical Scatter/Fluorescence Technology
- Light Scatter Principles
- Fluorescence Principles
Electrical Impedance Technology
‹ Absorption Spectrophotometry
Hemoglobin Measurement Process
Multi-Angle Polarized Scatter Separation
Single channel analysis of up to 20000 cells

900/ 90D0
0
7
Argon ion
laser light
00 00 : Size
70 : complexity
0 0 90 0 : lobularity
90 /90 D
0
0
7 90 D : granularity
Argon ion
laser light
00
 Differential Principles of
CELL-DYN 4000
Polynuclear Eosinophil

Mononuclear Neutrophil

Monocyte

Basophil

Lymphocyte
CELL-DYN 4000
CELL-DYN 4000
CELL-DYN 4000
CELL-DYN 4000
CELL-DYN 4000
CELL-DYN 4000
Fluorescence Flow Cytometry

FL1
530 nm
Green
RETC

FL3
630 nm
Red
NRBC,
Non-viable
WBCs
CELL-DYN 4000 : 특장점
 CELL-DYN 4000 NRBC Enumeration

• Red fluorescence is a measurement of the

amount of DNA stained by the dye.
• Axial light loss (0˚) measures cell size.
 NRBC Enumeration
NRBC Enumeration
Viable Non
WBCs viable
WBCs

NRBCs
 High NRBC Specimen: Neonate WBC DIFFERENTIAL LOW-
LOW-HI FL

CELL-DYN 4000
WBC: 7.27 109/L 0° 0°
NRBC: 9.71 NRBC/WBC: 134.
S S
I I
Microscopy Z Z
E E
NRBC/WBC: 150
Smudge Cells: +
7° COMPLEXITY FL3-DNA
FL3-

MGG STAIN 400 X MGG STAIN 1280 X

 Optical Configuration for
Reticulocyte Analysis

• Intermediate Angle
Forward Scatter, IAS,
(3 to 10°) - 세포내부의
그물구조 유무를 확인
•Green Fluorescence,
FL1, (515 to 545 nm) -
녹색형광발산 여부 확인
CD4K530-Stained Reticulocytes (400X)
 System Standardization for
Reticulocyte Analysis

FL1

IAS RETIC
RETIC RETIC Matured RBC
FL1 RNA

Reticulocytes Reticulocytes

IRF

7° FL1-RNA
Matured RBC
 Determination of White Cell Viability
Aged Blood CLL

Dead cells Smear cells

Fluorescence allows an accurate differential up to 36 hours.

Fluorescence provides additional information about aged samples

and in hematological malignancies.
 Malaria

Normal control Malaria

 Dual Angle Platelet Measurement

PSS (90o) PSS (90o)

RBC
Debris Argon laser Argon laser
light source light source IAS (7o)
IAS (7o)

Plts
Debris
Plt RBC

Excellent platelet / RBC separation

Platelet
Superior platelet/ /non
RBC separation
platelet separation
Platelet / Nonplatelet
High precision and linearity
separation
 CD61Immunoplatelet
CD61 Immunoplatelet Count
Count
IAS vs PSS (CD61) OPTICAL PLT
CELL-DYN 4000 LABORATORY WORKSHEET

PLT 193 PLTi 202

MPV 8.36 CD61 190 90° 90°
PDW 15.2 PLTs .20
PCT .161 PLTl .30

7° 7°
IMMUNO PLT CD61

F
L
1

C
D
6
1

7° FL1
 CD3/4/8 Immunoassay
CD3/4/8 Immunoassay
CD4+
CELL-DYN 4000 LABORATORY WORKSHEET
0 F
° I
Lymphocyte Subset Panel 1 T
C
WBC 11.9 10e3/µL WVF .984 S
I
LYM 1.60 10e3/µL %L 13.4 LVF .993 Z F
E L
1
T-Lymph Subsets
CD3+ 1.17 10e3/µL %C3+ 73.1 vC4+ 43 cells/µL 7° COMPLEXITY PE-FL2
CD4+ 48 cells/µL %C4+ 3.0 nvC4 5 cells/µL CD8+
CD4- 805 cells/µL %C4- 50.3 nv/v 0.1 cells/µL
CD8+ 816 cells/µL %C8+ 51.0 F
CD8- 46 cells/µL %C8- 3.0 I
T
R4:8 0.06 (ratio) C

F
L
1

PE-FL2