LAB METHODS FOR WBC EVALUATION

3.6-10.6×10^9/L

Normal Range of Leukocyte count

Monocyte, Neutrophil, Lymphocytes

3 part CBC only differentiates what?

5 part CBC

Differentiates only types of CBC

7 part CBC

includes all the types of WBC including the blast cells

Ethylenediamenetetraacetic acid (EDTA)

best specimen for all cell counts also for WBC evaluation

Blood from skin puncture

another specimen for WBC evaluation

1% hydrochloric acid

used to lyse RBCs (non-nucleated RBCs)

Within 30 minutes

until how long until nuclei of neutrophils swell, some loss of chromatin.
WBC morphology and deteriorates

Cytoplasmic vacuole

degenerative changes that appear in monocytes (mostly) and neutrophils.

• forms when there are foreign agents/antigen when there are infections

Nuclear lobulations (or Rieder cell)

degenerative changes in mononuclear cells

Tedious, Time consuming

MANUAL WBC COUNTING DISADVANTAGES

used when counts exceed instrument threshold
used when instrument is nonfunctional or no back-up
used in remote labs in 3rd world countries
used in case of a disaster when done in the field

MANUAL WBC COUNTING ADVANTAGES

Neubauer counting chamber

commonly used chamber

Hemacytometer/counting chamber

● For specimen with platelet counting interference

● Backup method

Used a check on the validity of electronic methods for calibration.

● As a check on the validity of electronic counts in

patients with profound leukopenia or

thrombocytopenia.

Thoma pipettes/Automated pipettes

Diluents

LOD

meaning limit of detection

LQD

limit of quantification

Open type
Closed type
Addis
Exton
Petroff

HEMACYTOMETRY ACCORDING TO TYPE

Spencer

Burker

Levy

Levy-Hausser

Open type of

Thomas-Zeiss

Closed type of

Thoma

Turk

Neubauer

Improved Neubauer

Fuchs-Rosenthal

Bass-Jones

ACCORDING TO RULINGS

LEVY CHAMBER WITH IMPROVED NEABAUER RULING

Composed of:
● 2 raised surfaces

• Each with 3mm x 3mm square counting

  area/grid separated by an H-shaped moat

  (Total area: 9mm^2)

● 9 1mm x 1mm squares

• Each of the 4 corner (WBC counting) squares is subdivided further into 16 squares

● A coverslip is placed on top of the counting surfaces.

● Distance between each counting surface and the coverslip is 0.1mm.

● Total volume of one entire grid or counting area on one side of the hemacytometer is 0.9mm3

● Hemacytometer and coverslips must meet the specifications of the National Bureau of Standards, as indicated by the initials “NBS” on the chamber