1) Hemoglobin (Hb)

Pre-Analytical Phase (Before Testing)

• Patient preparation: fasting, posture, timing (e.g., morning HbA1c)
• Sample collection: Correct tube (EDTA for CBC, fluoride oxalate for glucose, serum for biochemistry). Correct labeling (patient ID, date, time). Avoid hemolysis and clotted samples.
• Transport & storage: Maintain cold chain if needed (e.g., hormones). Test-specific storage temp (e.g., CBC within 6 hours).

Analytical Phase (Actual Testing)

• Use validated methods: Hematology: Automated analyzers, WHO-recommended cyanmethemoglobin method for Hb. Biochemistry: IFCC standard enzymatic methods for LFT, KFT, lipid profile. Immunology/Serology: ELISA, CLIA validated against WHO standards.
• Equipment calibration: Daily, weekly, monthly QC checks.

Post-Analytical Phase (After Testing)

• Verification of results before release.
• Delta check: Compare current and previous results for major shifts.
• Reporting: Clear, standard reference intervals, critical value alerts to physician.
• Record keeping: Maintain logs (instruments, QC, reports) for audit.

2) Red Blood Cells (RBC)

• Specimen: Whole blood (2–3 mL) collected in EDTA (lavender-top tube).
• Precautions: Avoid clotted samples. Mix gently after collection (do not shake). Process within 2–4 hours; if delayed, refrigerate at 2–8 °C (not >24 hrs).

Automated Analysis

• Load sample into hematology analyzer.
• Obtain RBC, Hb, Hct, indices, WBC count, and preliminary differential.

Manual Methods

• RBC Count: Dilute blood with RBC diluting fluid, charge Neubauer chamber, count under microscope.

3) White Blood Cells (WBC)

• Sample type: Whole blood (EDTA anticoagulated).
• Volume: 2–3 mL (minimum 1 mL for pediatric).
• Anticoagulant: EDTA (lavender-top tube).
• Precautions: Avoid clotted samples. Mix gently (do not shake vigorously). Process within 2 hours of collection (max 6 hours at room temp, 24 hours at 2–8°C).

(A) Automated Hematology Analyzer (Preferred)

• Uses electrical impedance or flow cytometry.
• Provides Total WBC Count + DLC (neutrophils, lymphocytes, monocytes, eosinophils, basophils).
• Internal QC and calibration required daily.

(B) Manual Method

• Diluting fluid: Turk’s solution. Instrument: Neubauer counting chamber.
• Procedure: Dilute blood (1:20), charge chamber, allow cells to settle, count WBCs, calculate WBC/µL.

Safety & Biosafety

• Use PPE (gloves, coat). Handle blood as per universal precautions. Dispose waste as per biomedical waste rules.

4) Blood Pressure

Equipment

• Standard mercury sphygmomanometer, Aneroid sphygmomanometer (calibrated), Digital/automatic BP monitor (validated), Stethoscope.
• Appropriate cuff sizes: Adult standard, Large adult, Small adult/pediatric.

Pre-Measurement Preparation

• Patient should rest 5 minutes, avoid caffeine/smoking/exercise 30 min prior, sit quietly, arm at heart level. Correct cuff size.

Manual (Auscultatory) Method

• Wrap cuff, palpate brachial artery, inflate 20–30 mmHg above disappearance of radial pulse, deflate slowly 2–3 mmHg/sec.

Digital (Oscillometric) Method

• Position cuff, start device, record systolic, diastolic, pulse rate.

Quality Control

• Aneroid & digital devices calibrated every 6–12 months. Staff trained annually.

5) Pulse Rate

• Site: Radial artery, carotid, brachial, or apical.
• Position: Patient at rest, sitting or lying.
• Procedure: Use index/middle finger, count 30 sec × 2 (if regular) or 1 min (if irregular). Record rate, rhythm, character.
• Preconditions: Rested 5 min, no recent exertion, caffeine, smoking.
• Check at baseline, pre-anesthetic, re-check if abnormal.

6) Respiratory Rate

• Patient at rest, unaware of observation. Observe chest/abdomen rise and fall. Count full minute. Record rate, rhythm, depth, effort.
• Preconditions: Rested 5 min, avoid exertion/caffeine/smoking.

7) Spirometry

• Equipment: Spirometer (ATS/ERS approved), mouthpieces, calibration syringe, software.
• Patient: No smoking/alcohol/caffeine, avoid bronchodilators, loose clothes, rest 10–15 min. Contraindications checked.

Calibration & QC

• Daily calibration with 3L syringe. Logs maintained.

Test Steps

• Explain procedure, seat upright, nose clip, inhale fully, exhale hard/fast till empty. Repeat 3 acceptable maneuvers (max 8). Criteria: reproducibility within 150 mL.

Infection Control

• Disposable mouthpieces or sterilize. Clean equipment daily.

8) Height

• Equipment: Stadiometer. Patient barefoot, heels together, back straight, head in Frankfurt plane. Lower headpiece, read nearest 0.1 cm.

9) Weight

• Equipment: Digital scale. Flat surface. Patient removes shoes/clothes, stands centered. Record nearest 0.1 kg.

10) Body Mass Index (BMI)

• Calculation: BMI = weight (kg) / height (m²).

11) Waist–Hip Ratio

• Equipment: Non-stretchable tape.
• Waist: Midpoint between rib and iliac crest, end of expiration. Hip: Widest buttocks. Record to 0.1 cm. W/H ratio = WC ÷ HC.
• QC: Take twice, average. Same observer if possible.

12) Random Blood Sugar (RBS)

• Specimen: Venous blood preferred. Anticoagulant: Sodium fluoride + potassium oxalate. Stable 24 hr at 2–8 °C. Reject hemolyzed/unlabeled.
• Methods: GOD-POD, Hexokinase, GDH. POC: Glucometer (cross-check abnormal).

13) Blood Groups

• Specimen: EDTA blood or clotted. Storage: 2–8 °C (≤48 hr). Reject hemolyzed/clotted/mislabeled.
• ABO: Forward grouping (cells with antisera), Reverse grouping (serum with A/B cells).
• Rh: Cells with anti-D reagent. Agglutination = Rh+, none = Rh–.
• QC: Validated antisera, daily QC, reagent storage 2–8 °C, EQAS participation.