Automated Blood Cell Analyzer

History
• First period (2217-2417 BE) - Microscope inventio
n, Manual blood count
• Second peroid (2418-2495 BE) - Manual CBC intro
duced
• Third period (2496-2529 BE) - Automation introd
uced
• Fourth period (2530-present) - Advanced automati
on introduced
• Fifth period (present and on) - Get benefit from t
echnology
 Major principles
1. Electrical impedance
2. Light scattering
Electrical Impedance
Light Scattering
 Parameters generated from
Automated Blood Analyzers
• Hb • Total WBC count
• Hct • WBC Diff count
• RBC count • Plt count
• MCV • PDW
• MCH • MPV
• MCHC • Reticulocyte count
• CHCM
• HDW
Hemoglobin concentration (Hb)

Techniques
1. Cyanmethemoglobin method
2. High-angle light scattering method

Application
Evaluate anaemic status
 RBC count
Techniques
1. Electrical impedance
2. Light scattering

Applications
1. Evaluate anaemic stutus
2. Evaluate polycythemic status
 Mean Corpuscular Volume (MCV)
Techniques
1. Electrical impedance : Height of
pulse obstructed--> Mean volume
distribution
2. Light scattering : Low-angle (2-3o)
light scattering --> Mean size
distribution
3. Calulation : Hct x 10
RBC
Applications
Evaluate hypochromic microcytic anaemia
 Hematocrit (Hct)
Techniques
1. Calculation : MCV x RBC
10
2. Directly obtained : Mean pulse

Applications
Evaluate anaemic and polycythemic st
atuses
Mean Corpuscular Haemoglobin (MC
H)
Techniques
Calculation : Hb x 10
RBC
Application (NR 29.5+/-2.5pg)
Evaluate microcytic hypochromic
anaemia
Mean Corpuscular Haemoglobin Con
centration (MCHC)
Technique
Calculation : Hb x 100
Hct

Application (NR 33+/-1.5 g/dl)

Evaluate dehydration state of RBC
High in HS
Corpuscular Haemoglobin Concentra
tion Mean (CHCM)
Technique
Directly obtained (Only from light
scattering) : High-angle (5-15o)
light scattering

Application (NR ?)
Still to be determined
Haemoglobin Distribution Width (H
DW)
Technique
-SD of Hb concentration distribution h
istogram
-Only obtained from Laser technology

Application (NR < 3 g/dl)

-Determine Hb-content heterogeniety
-Evaluate dehydration state of RBC
 Red Cell Distribution Width (RDW)

Techniques
Calculation
RDW-CV = SD x 100
MCV
RDW-SD = SD of RBC histogram
Application (NR 12.8 +/-1.2%)
-Determine anisocytosis
-Evaluate hypochromic microcytic anaemia
 Total WBC count
Techniques
1. Eletrical impedance : No of pulse
obstructed
2. Light scattering : No of signal from
low-angle light scattering
Applications
Evaluation of leukopenia and leukocytos
is
 WBC differential count
Techniques
1. By volume
2. By volume + conductivity + light
scattering
3. By light scattering + cytochemistry
4. By light scattering + RF impedance
Types
3-part, 5-part, 6-part, 7-part
Three-part diff histogram
Five-part diff histogram
Six-part diff histogram
Mean Peroxidase Activity Index (MPXI
)

• Derived by cytochemistry-based te
chnique (Technicon)
• Increased in bacterial, PV, PF infec
tions
• Unchange in viral infection
 Platelet count

Techniques
1. Electrical impedance : Volume cut-of
f at 2fl-20fl
2. Light scattering : High-angle light s
cattering for size and Hb content
Application
Evaluation of thrombocytosis and thr
ombocytopenia
Platelet histogram
 Platelet Distribution Width
(PDW)
Techniques
Calculation
PDW-CV = SD x 100
MPV
Application (NR < 20%)
1. High : AA, MA, CML, on
chemotherapy
2. Low : still in observations
 Mean Platelet Volume (MPV)
Techniques
1. Calculation : Mean volume of platelet
in fitted curve
2. Calculation : Mode of platelet volume
distribution
Applications (NR 9.0-9.8 fl)
1. High : ITP, Post splenectomy, DIC
2. Low : AA, MA, post chemotherapy
 MPV nomogram
MPV
 Reticulocyte count
Techniques
1.Fluorescence detection of red cells s
tained with RNA specific fluorochro
ms ( Auramine O, Ethidium bromide,
Oxazine 750, Thiazole orange)
2. Direct count via volume, light scatte
ring and opacity of cells
 Reticulocyte count

Types after fluorescence detection

1. Low Fluorescence Ratio (LFR): Most
mature
2. Middle Fluorescence Ratio (MFR):
mid mature
3. High Fluorescence Ratio (HFR):
Most immature
 Reticulocyte count

Application (NR 0.5-2.5%)

Evaluation of BM efficiency
RBC maturation
Maturation time of reticulocytes
 NRBC enumeration
• Stain DNA with fluorescence dye (polymethine)
and measure for size and intensity by light
scattering to differentiate NRBC from WBC
(Sysmex XE-2100, Abbott CELL-DYN 4000,
Abbott CELL-DYN Sapphire)
• Detect NRBC under lymphocyte region in WBC
volume/scatter plot and count NRBC by
extrapolation of low-volume peak of WBC
distribution curve (Beckman Coulter LH750)
 Some Automated Blood Analyzers
in Today Market

• Beckman-Coulter (MAXM, STKS, LH) : VCS

• Sysmex : EI + LS + RF impedance
• Abbott CELL-DYN : EI + LS + MAPSS (Multi
Angle Polarized Scatter Separation) +
Fluorescence staining
• Bayer Advia : LS + Cytochem
 References
1. ธานินทร์ อินทรกำธรชย ั (บรรณาธิการ) โลหิตวิทยาในเวชปฏิบต ั .ิ
กรุงเทพ :บริษัท บียอนด์ เอนเตอร์ไพรซ ์ จำกัด, 2542.
2. Dacie JV, Lewis SM. Practical Haematology (8th edition).
London : Churchill Livingstone, 1995.
3. Koepke JA. Practical Laboratory Hematology. New York :
Churchill Livingstone, 1991.
4. Turgeon ML. Clinical Hematology (3rd Edition) : Theory and Procedures.
Philadelphia : Lippincott Williams & Wilkins ; A Wolters Kluwer Company, 1999.
5. Hall R & Malia RG. Medical Laboratory Haematology (2nd Edition).
Oxford : Butterworth-Heinemann Ltd, 1991.
6. นวพรรณ จารุรักษ์ . กรณีศก ึ ษาทางโลหิตวิทยา: การตรวจวิเคราะห์เลือดด ้วย
เครือ่ งอัตโนมัตท ้ กริ ย
ิ ใี่ ชปฎิ ิ าเคมี. กรุงเทพ : หจก โรงพิมพ์ชวนพิมพ์, 2540.
7. นวพรรณ จารุรักษ์ . เครือ ่ งวิเคราะห์เม็ดเลือดอัตโนมัต ิ : เทคโนโลยีแห่งปั จจุบน
ั และอนา
วารสารโลหิตวิทยาและเวชศาสตร์บริการโลหิต 2545; 12 : 309-329.