CL03 - Laboratory Test of RBC: Principle & Interpretation

Sunday, December 21, 2014

5:25 PM

Outlines

Blood smear examination

Complete blood count (CBC)

RBC Indices, Reticulocyte count & Reticulocyte Production Index
Blood smear examination
Laboratory diagnosis of anemia

glass slide -->

spreader

film

oil (

40

area

cell

Complete Blood Count (CBC)

cell

Classification

Alpha-thalassemia
(HbH disease)

RBC indices

RBC : RBC count

Hb : Hemoglobin content (g/dl)
Hct: Hematocrit (%)

RBC
MCV : Mean Corpuscular Volume (fl)

80-100 fl : Normocytic
< 80 fl : Microcytic
> 100 fl : Macrocytic

Hb conc.
varies depending on age, sex, race

Cause of Anemia

(Increase destruction)

(Excessive loss of RBC)

(Decrease RBC production)

morphology

Special Staining Inclusions

RBC Indices
2.1

-->

Suitable area for observation

Estimate cell numbers
WBC: 10-20/LPF ( 40X)
Platelets: 7-20/OF ( 100X)
Determine predominant cell populations
Examine cellular morphology / Differential WBC count

Abnormal RBC Morphology

2

spherocyte
Hb

1. WBC Count
2. WBC differential

blood smear
3. RBC indices

4. Platelet count and volume

platelet (
RBC)
5. Blood smear evaluation:

blood smear
morphology
platelet

Laboratory Diagnosis of Anemia

Anemia

(Is a sign, not a disease)

Reduced RBC number, Hb or Hct

Cell

positive

inclusion bodies
RBC

RBC)

Cytometric Classification (

Inclusion Bodies
plasma

%
Immature RBC in
peripheral blood

Reticulocyte
RNA
80

MCV
80
...
MCH : Mean Corpuscular Hemoglobin (pg)

hemoglobin
MCHC : Mean Corpuscular Hb Concentration (g/dl)

Hb
RDW : Red Cell Distribution Width (%)

(Anisocytosis)
*Anisocytosis :
**Poikilocytosis :
RDW
iron def. anemia
RDW
anemia

Thalassemia
RDW

2.2

RBC Morphology

G6PD

RBC

oxidative stress of Hb

(II) Hypochromic Microcytic Anemia

Thalassemia
Iron deficiency

--> Hb

Wright's stain

(III) Normochromic Macrocytic Anemia

Vit B12 def
Folate def

Size

Color

Inclusions & Parasites

Nucleated RBC
2

1.1) Anisocytosis

RNA
mature RBC
anemia

Erythrokinetic Classification

RBC

reticulocyte

RNA

RBC
cytoplasm

Normal value
Adult 0.5 2.0 %
Newborn 2.5 - 6.5 %

Thalassemia
Heavy metal poisoning

Variation in Size of RBC

Normocyte, Microcyte, Macrocyte

Corrected Reticulocyte Count (Reticulocyte Index)

Corrected for degree of anemia

% reticulocyte -->
%
RBC

anemia
loss RBC
-->
% reticulocyte
correct
reticulocyte count

RBC

cytoplasm
)
small lymphocyte

Erythrocyte distribution

(I) Normogenerative anemia

RBC

(II) Hyporegenerative anemia

RBC

(III) Thalassemia major

Hct

RBC

Lab. Test for RBC Turn Over

Reticulocyte count

RBC

multiple myeloma
Increased in fibrinogen or globulin

reticulocyte

8%

RBC
(

> 1/3

(Normal absolute RC = 60,000 100,000 / ml)

Correct RC = (patientsHct/45%) x RC
= (25%/45%) x 8% = 4.4%

RBC
neutrophil
orthochromatic
nucleus

Small lymphocyte

Ex
Normal person : 5 million RBC, 45%Hct, 1%RC = 50000 retic.
Anemia person : 3 million RBC, 25%Hct, 8%RC = 240000 retic.

2.4

MCHC), Normocytic (
MCV) Anemia
(I) Normochromic (

Chronic disease
Acute blood loss

1
G6PD Def.

Reticulocyte count

Reticulocyte

-->

1000 RBC

2.3

RBC

mature

wrights stain

i. Reticulocyte count

Inclusions & Parasites

Reticulocyte Production Index (RPI)

RBC nucleus
orthonormoblast (
--> exclude nucleus
reticulocyte
Reticulocyte
2-3
blood circulation

blood circulation 1
mature RBC
-->
RBC

-->
reticulocyte

blood circulation
= maturation time
(maturation time
reticulocyte

hemoglobin (
+ RNA(

)
)

RBC

RBC

mature RBC)

nuclear remnant

!!

central pallor

ii. Serum unconjugated bilirubin/

urine urobilinogen concentration

Ag-Abs reaction
Autoimmune HA

RPI > 2.0 Normal BM response (

RPI < 2.0 Inadequate BM response

RBC

Severe anemia
Hemolytic anemia
Splenectomy

Shape
**Poikilocytosis = Variation in Shape of RBC

iii. Serum haptoglobin concentration

fibrous tissue
RBC

Hereditary spherocytosis
Hemolytic anemia

fragmented red cell

RBC

Hereditary stomatocytosis
Alcoholic Cirrhosis
Lead poisoning
Neoplastic disease

burn

Hemolytic anemia
Severe burns
DIC

Thalassemia
Myelofibrosis
Hemolytic anemia

hemolytic anemia
Alcoholic liver disease
Lipid metabolism
disorder
Splenectomy
3

Biochemical Classification
Depleted cofactor necessary for normal Hematopoiesis
(iron, ferritin, folate, VitB12)

Blood transfusion

Abnormal enzyme function

(G6PD, pyruvate kinase)

Elliptocyte
Ovalocyte

Hereditary elliptocytosis
Hemolytic Anemia

Lecture Page 1

Thalassemia
Sickle cell anemia
Liver disease

Abnormal function of immune system (Coombs test)

RBC
oxygen
hemoglobin

oxygen
reversible

Sickle cell anemia

Homozygous HbS

Microangiopathic
Hemolytic Anemia
Glomerulonephritis

acantocyte
artifact

slide

Liver disease
Malignant hypertension
Carcinoma
Infectious disease

 Hematocrit
conc. of RBC, not total RBC mass

1: normal
Hct

40-50%

2: anemia

Hct
acute blood loss

3:

plasma

anemia
chronic disease
RBC
plasma
volume
RBC

-->
Note :

Hct
red count, Hct,

Hb

4:

anemia
Hct
dengue

Initial Investigation for Anemia

Patient history/ physical examination
CBC : RBC indices anemia?, type?
Blood smear aniso-poikilocytes?
Reticulocyte count marrow response

Differential diagnosis

Laboratory examination

Anemia : RBC, Hb, Hct

Classify type : MCV, MCH, MCHC
Anisocytosis : RDW
BM response : Reticulocyte count

Case 1
Anemia ?
Anemia
Type ?
Microcytic hypochromic
Differential diagnosis ?
Iron def. a. Thalassemia
Iron studies Hb typing
serum iron, ferritin : low
TIBC : high

Case 2
Anemia ?
Anemia

Type ?
Microcytic hypochromic
Differential diagnosis ?
Iron def. a. Thalassemia
Iron studies Hb typing
Hb F = 2 %
Hb A2 = 5 %

Lecture Page 2

Interpretation of iron studies