Automated Blood Cell Analyzer

History• - 22172417First period ( BE) - M MMMMMMM

MMMMMMMMMM M MMMMM MMMMM MMMMM ,• - (2418 2495 ) - Manual CBC introduced• - (2496 2529 ) - MMMMM MMMMM MMMMMMMMMM• -Fourth period (2 5 3 0 present) - Advanced a

utomation introduced• MMMMMMMM MMM MMM() - M MM MMMMMMM MMMM MMMMMMMM

gy

Major principles

1. Electrical impedance2. Light scattering

Electrical Impedance

Light Scattering

Parameters generated from Automated Blood Analyzers

• Hb• Hct• RBC count• MCV• MCH• MCHC• CHCM• HDW

• Total WBC count• WBC Diff count• Plt count• PDW• MPV• Reticulocyte count

Hemoglobin conc entration (Hb)

Techniques 1. Cyanmethemoglobin method

- 2. High angle light scattering method

MMMMMMMMMMM Evaluate anaemic status

RBC countTechniques

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 MMMMM MMMMMMMMMM--M M MMM MMMMM M MMMMMMMMMMMM - -2 23. Light scattering : Low angle ( o ) -- l i ght scat t er i ng >Mean si ze MMMMMMMMMMMM 3. Calulation : MMM 10x RBCApplications Evaluate hypochromic microcytic anaemia

Hematocrit (Hct)

Techniques 1. Calculation : MMM M MMM

2. Directly obtained : Mean pulse

Applications Evaluate anaemic and polycythemic

statuses

10

Mean Corpuscular Haemoglobin (MCH)

Techniques Calculation : Hb 10 MMM

-Application (NR 29.5+/ 2.5pg) Evaluate microcytic hypochromic MMMMMMM

Mean Corpuscular Haemoglobin Concentration (MCHC)

Technique Calculation : Hb 100 Hct

Application - (NR 33+/ 1.5 g/dl) Evaluate dehydration state of RBC High in HS

Corpuscular Haemoglobin Conce ntration Mean (CHCM)

Technique Directly obtained (Only from light scattering) : High-angle (5-15o) light scattering

Application (NR ?) Still to be determined

Haemoglobin Distribution Width (HDW)

Technique - SD of Hb concentration distribution h

istogram - Only obtained from Laser technology

3Application (NR < g/dl) - - Determine Hb content heterogeniety - Evaluate dehydration state of RBC

Red Cell Distribution Width (RDW)

Techniques Calculation

- RDW CV = SD 100x 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 MMMMMMMMMM 2 . :Light scattering No of signal from MMM -MMMMM MMMMM MMMMMMMMMMApplications MMMMMMMMMM MM MMMMMMMMMM MMM MMMMMMM

MMMMM

WBC differential count

Techniques 1. By volume 2 . + +By volume conductivity light MMMMMMMMMM 3 . +By light scattering cytochemistry 4 . +By light scattering RF impedanceTypes - - - -3 5 6 7part, part, part, part

- Three part diff histogram

- Five part diff histogram

- Six part diff histogram

Mean Peroxidase Activity Index (MPXI)

• - Derived by cytochemistry based technique (Technicon)

• MMMMMMMMMM MMM MM MMMM, ,ctions

• MMMMMMMM MM MMMMM MMMMMMMMM

Platelet count

Techniques 1. Electrical impedance : Volume c

- -ut off at 2fl 20fl - 2. Light scattering : High angle ligh

t scattering for size and Hb contentApplication Evaluation of thrombocytosis and t

hrombocytopenia

Platelet histogram

Platelet Distribution Width (PDW)

Techniques Calculation PDW-CV = SD x 100 MPVApplication (NR < 20%)1. High : AA, MA, CML, on

chemotherapy2. Low : still in observations

Mean Platelet Volume (MPV)

Techniques 1. Calculation : Mean volume of platele

M MM MMMMMM MMMMM 2 . :Calculation Mode of platelet volum

M distribution

- 9098Applications (NR . . fl) 1. High : ITP, Post splenectomy, DIC 2. Low : AA, MA, post chemotherapy

MPV nomogram

MPV

Reticulocyte count

Techniques1. Fluorescence detection of red cel

ls stained with RNA specific fluoroc hroms ( Auramine O, Ethidium bro

mide, Oxazine 750, Thiazole orange)

2. Direct count via volume, light sc attering 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)

MMM M MMMMM MMMM MMMMM MMMMMMMMMMM T oday Market

• MMMMMMM-MMMMMMM MMMMMM MMMM, LHM M MMM• Sysmex : EI + LS + MM MMMMMMMMM• Abbott CELL-DYN : EI + LM + MAPSS

(Multi Angle Polarized Scatter Separation) + Fluorescence staining

• Bayer Advia : LS + Cytochem

1. ธานิ�นิทร์ อิ�นิทร์กำ าธร์ชั�ย (บร์ร์ณาธ�กำาร์ ) โลหิ�ตวิ�ทยาในิเวิชัปฏิ�บ�ต� . กำร์�งเทพ : บร์�ษั�ท บ ยอินิด์ เอินิเตอิร์ไพร์ซ์ จำ ากำ�ด์ , 2542.

2 8. Dacie JV, Lewis SM. Practical Haematology ( th edition). 1995London : Churchill Livingstone, .

3. Koepke JA. Practical Laboratory Hematology. New York : ,1991.

4 3. Turgeon ML. Clinical Hematology ( rd Edition) : Theory and Procedures. 1999Philadelphia : Lippincott Williams & Wilkins ; A Wolters Kluwer Company, .

5 2. Hall R & Malia RG. Medical Laboratory Haematology ( nd Edition). - :, 1991.

6. นิวิพร์ร์ณ จำาร์�ร์�กำษั . กำร์ณ ศึ&กำษัาทางโลหิ�ตวิ�ทยา : กำาร์ตร์วิจำวิ�เคร์าะหิเล)อิด์ด์*วิยเคร์)+อิงอิ�ตโนิมั�ต�ท +ใชั*ปฎิ�กำ�ร์�ยาเคมั . กำร์�งเทพ : หิจำกำ โร์งพ�มัพชัวินิพ�มัพ 2540, .

7. นิวิพร์ร์ณ จำาร์�ร์�กำษั . เคร์)+อิงวิ�เคร์าะหิเมั.ด์เล)อิด์อิ�ตโนิมั�ต� : เทคโนิโลย แหิ0งป1จำจำ�บ�นิและอินิาคต . วิาร์สาร์โลหิ�ตวิ�ทยาและเวิชัศึาสตร์บร์�กำาร์โลหิ�ต -2545 12 309329; : .

References