Perfect 👍.

You’ve shared another image that lists Hematological tests for the diagnosis of
blood disorders. I’ll draft a long, detailed explanatory note based on the standard reference
Harsh Mohan – Textbook of Pathology (recent edition, e.g., 8th/9th edition), suitable for
medical students.

🩸 Hematological Tests for the Diagnosis of Blood Disorders

The hematopoietic system is frequently affected in systemic as well as primary hematological

diseases. Laboratory investigations are critical for diagnosis, monitoring, and prognosis of
these conditions. According to Harsh Mohan (Textbook of Pathology, latest edition),
hematological tests can be grouped into blood tests, coagulation tests, and blood chemistry
tests.

1. Blood Tests
These are the most basic and essential investigations:
 Hemoglobin (Hb) estimation
o Normal values:

 Male: 13–17 g/dL

 Female: 12–15 g/dL
o ↓ Hemoglobin → anemia (nutritional, hemolytic, blood loss).

o ↑ Hemoglobin → polycythemia (primary or secondary).

o Methods: Cyanmethemoglobin method (standard), automated analyzers.

 White Blood Cell (WBC) count

o Total Leukocyte Count (TLC):

 Normal: 4,000–11,000 /mm³.

 Leukocytosis → infections, leukemia, stress.
 Leukopenia → marrow failure, viral infections, drugs.
o Differential Leukocyte Count (DLC):

 Neutrophils, lymphocytes, eosinophils, basophils, monocytes.

 Helps in detecting acute bacterial infection (neutrophilia), viral
infection (lymphocytosis), allergy/parasitic infection (eosinophilia).
 Platelet count
o Normal: 150,000–450,000 /mm³.
 o Thrombocytopenia → bleeding disorders (ITP, marrow suppression).

o Thrombocytosis → myeloproliferative disorders, reactive states.

 Packed Cell Volume (PCV/Hematocrit)

o Proportion of blood volume occupied by RBCs.

o Normal:

 Male: 40–50%
 Female: 36–44%
o ↓ PCV → anemia.

o ↑ PCV → polycythemia.

 Erythrocyte Sedimentation Rate (ESR)

o Normal: Westergren method –

 Male: 0–15 mm/hr

 Female: 0–20 mm/hr
o ↑ ESR in chronic infections, autoimmune diseases, malignancies.

o Nonspecific but useful in monitoring disease activity (e.g., TB, rheumatoid

arthritis).

2. Coagulation Tests
These assess the hemostatic mechanism of blood:
 Bleeding Time (BT)
o Measures platelet function and capillary integrity.

o Normal: 2–7 minutes (Ivy method).

o Prolonged in platelet disorders (e.g., ITP), von Willebrand disease.

 Prothrombin Time (PT)

o Tests extrinsic and common coagulation pathways (factors II, V, VII, X,
fibrinogen).
o Normal: 11–16 seconds.

o Prolonged in vitamin K deficiency, liver disease, warfarin therapy.

o Used to calculate INR (international normalized ratio).

 Activated Partial Thromboplastin Time (APTT)

 o Tests intrinsic and common coagulation pathways (factors VIII, IX, XI, XII).

o Normal: 25–40 seconds.

o Prolonged in hemophilia A/B, heparin therapy, disseminated intravascular

coagulation (DIC).
 Clotting Time (obsolete, rarely used now)
o Rough test of overall coagulation, replaced by PT/APTT.

3. Blood Chemistry Tests

These are supportive but often necessary for complete evaluation of hematological disorders:
 Serum Iron, Ferritin, TIBC (Total Iron Binding Capacity) → assessment of iron
deficiency anemia.
 Vitamin B12 and Folate levels → megaloblastic anemia.
 Serum Bilirubin, LDH, Haptoglobin → indicators of hemolysis.
 Uric Acid → raised in increased cell turnover (e.g., leukemias, myeloproliferative
disorders).
 Serum Protein Electrophoresis → diagnosis of plasma cell dyscrasias (e.g., multiple
myeloma).

📚 References

1. Mohan, Harsh. Textbook of Pathology, 9th Edition, Jaypee Brothers Medical

Publishers, New Delhi, 2022.
2. Kumar, V., Abbas, A.K., Aster, J.C. Robbins and Cotran Pathologic Basis of
Disease, 10th Edition, Elsevier, 2020.
3. Hoffbrand, A.V. Essential Haematology, 7th Edition, Wiley-Blackwell, 2016.

✅ This note is structured for MBBS-level pathology learning and highlights the normal
values, diagnostic significance, and clinical utility of hematological investigations.
Would you like me to also create a tabular comparison (test → normal range → clinical
significance) for quick exam revision?
Great 👍 A tabular comparison will make this much easier for exam prep and quick recall.
Here’s a structured summary table based on Harsh Mohan – Pathology (latest edition) and
other standard references:

🩸 Hematological Tests – Quick Revision Table

Test Normal Range Clinical Significance

↓ Anemia (iron deficiency, hemolytic,

Hemoglobin (Hb) ♂: 13–17 g/dL♀: 12–15 g/dL blood loss)↑ Polycythemia (primary or
secondary)

↑ Leukocytosis (infection, leukemia)↓

Total Leukocyte
4,000–11,000 /mm³ Leukopenia (viral infection, marrow
Count (TLC)
suppression, drugs)

Neutrophils 40–
Pattern useful in diagnosis:Neutrophilia
Differential 70%Lymphocytes 20–
→ bacterial infectionLymphocytosis →
Leukocyte Count 40%Eosinophils 1–
viral infectionEosinophilia → allergy,
(DLC) 6%Monocytes 2–
parasites
10%Basophils 0–1%

↓ Thrombocytopenia (ITP, marrow

failure)↑ Thrombocytosis
Platelet Count 150,000–450,000 /mm³
(myeloproliferative disorders, reactive
states)

Packed Cell Volume

♂: 40–50%♀: 36–44% ↓ Anemia↑ Polycythemia
(PCV / Hematocrit)

Erythrocyte ↑ Chronic infections (TB), autoimmune

♂: 0–15 mm/hr♀: 0–20
Sedimentation Rate diseases, malignanciesNonspecific but
mm/hr
(ESR) useful for disease monitoring

Prolonged in platelet function defects

Bleeding Time (BT) 2–7 min (Ivy method)
(e.g., ITP, von Willebrand disease)

Prothrombin Time Prolonged in vitamin K deficiency, liver

11–16 sec
(PT) disease, warfarin therapy

Activated Partial
Prolonged in hemophilia A/B, heparin
Thromboplastin 25–40 sec
therapy, DIC
Time (APTT)

↓ Iron deficiency anemia↑ Hemolytic

Serum Iron 60–170 µg/dL
anemia, hemochromatosis
Test Normal Range Clinical Significance

↓ Iron deficiency↑ Chronic disease, iron

Ferritin 15–300 ng/mL
overload

Vitamin B12 200–900 pg/mL ↓ Megaloblastic anemia

Folate 3–17 ng/mL ↓ Megaloblastic anemia

↑ Hemolysis (unconjugated), liver

Serum Bilirubin Total: 0.3–1.2 mg/dL
disease

LDH 140–280 U/L ↑ Hemolysis, leukemias

♂: 3.5–7.2 mg/dL♀: 2.6–6.0 ↑ Leukemias, myeloproliferative states,

Uric Acid
mg/dL gout

✅ This table format is high-yield for exams and helps in differential diagnosis at the
bedside.
Would you like me to also prepare a flowchart of investigations for anemia and bleeding
disorders (stepwise diagnostic approach) as an additional visual aid?
Excellent question 👍. The colour of the blood collection vial (vacutainer tube) depends on
the anticoagulant or additive present inside, which determines the type of blood test it is
used for. Using the wrong vial can lead to erroneous results.
Here’s a detailed guideline with references (based on Harsh Mohan – Pathology, CLSI
standards, and WHO guidelines):

🧪 Blood Collection Vials and Their Uses

Tube Cap
Additive/Anticoagulant Common Tests Guidelines / Notes
Colour

- Complete Blood
Count (CBC)- Prevents clotting by
EDTA Hemoglobin, chelating calcium. Fill 2/3
Purple /
(Ethylenediaminetetraacetic Hematocrit- to 3/4 full. Mix gently.
Lavender
acid) Peripheral smear- Avoid clotting or dilution
ESR (Wintrobe errors.
method)

Blue Sodium Citrate (3.2% or - Coagulation profile Ratio is critical → 1 part

(Light 3.8%) (PT, INR, APTT, D- citrate : 9 parts blood.
Blue) dimer, Fibrinogen)- Underfilling → falsely
Tube Cap
Additive/Anticoagulant Common Tests Guidelines / Notes
Colour

ESR (Westergren
prolonged clotting times.
method)

- Serum chemistry Blood clots → serum is

(LFT, KFT)- separated after
No anticoagulant (may have
Red / Plain Hormones, serology centrifugation. Allow 20–
clot activator)
(HIV, HBsAg)- Drug 30 min for clotting before
levels centrifuge.

- Biochemistry Gel barrier separates

Yellow (serum chemistry serum from clot →
(Sterile, Gel separator + Clot activator panels)- prevents contamination.
SST tube) Immunology, Preferred for many
serology automated analyzers.

- Plasma chemistry Heparin inhibits thrombin.

(electrolytes, liver Used where plasma is
Heparin (Lithium/ Sodium/
Green function, renal required. Avoid for
Ammonium Heparin)
function)- hematology (causes
Cytogenetics leukocyte clumping).

Fluoride stabilizes glucose

- Blood glucose
Sodium Fluoride + Potassium up to 24 hrs. Not suitable
Gray (inhibits glycolysis)-
Oxalate for calcium or enzyme
Lactate levels
assays (inhibitory effects).

Larger bore tube. Strict

Sodium Citrate (different - ESR (Westergren ratio of
Black
concentration, usually 3.8%) method) blood:anticoagulant
required.

Yellow Prevents complement

SPS (Sodium Polyanethol - Blood culture
(Culture activation & phagocytosis,
Sulfonate) (microbiology)
Bottle) supports bacterial growth.

- Trace element Free from metal

Specially treated (EDTA /
Royal Blue analysis (Zinc, contamination. Avoids
Heparin / None)
Copper, Lead) trace element errors.

Brown / - Lead level Certified lead-free to

EDTA (lead-free)
Tan estimation prevent contamination.

✅ Guidelines for Proper Use

 1. Order of Draw (to prevent cross-contamination)
o 1st: Blood culture vial (yellow, SPS).
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o 2nd: Coagulation tube (light blue).

o 3rd: Serum tube (red/yellow with gel).

o 4th: Heparin tube (green).

o 5th: EDTA tube (purple).

o 6th: Fluoride tube (gray, glucose).

(As per CLSI & WHO recommendations)

2. Mixing: Tubes with anticoagulants must be gently inverted 8–10 times after
collection (never shaken) to avoid clotting/hemolysis.
3. Volume: Always fill tubes to the recommended mark, especially citrate tubes
(PT/APTT) → underfilling alters plasma-anticoagulant ratio.
4. Transport & Storage:
o CBC → test within 2–4 hrs at room temperature.

o Coagulation → test within 1 hr (keep at room temperature).

o Glucose → fluoride tube stable up to 24 hrs.

📚 References

1. Mohan, Harsh. Textbook of Pathology, 9th Edition, Jaypee Brothers, New Delhi,
2022.
2. CLSI (Clinical and Laboratory Standards Institute). Collection of Diagnostic Venous
Blood Specimens, CLSI GP41-A7, 7th Edition.
3. WHO. WHO Guidelines on Drawing Blood: Best Practices in Phlebotomy,
Geneva, 2010.

👉 Would you like me to prepare a colour-coded chart/diagram (visual infographic) of these

vials so you can quickly revise them before exams and in the lab?

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