INSTRUMENTS

Vacutainers
• Types of blood samples:
• Whole blood: CBC, Glucose, urea, Coombs’
test
• Serum: Biochemical tests, Antibody testing
• Plasma: Coagulation studies, Factor assays
• EDTA: Chelation of Calcium ions
• Sod. Citrate: Trisodium citrate pentahydrate,
3.8% solution used. Loosely binds calcium ions
and creates Calcium citrate.
• 1:9 for coagulation studies
• 1:4 for ESR (0.4ml citrate+1.6 ml blood)
• Double oxalate: Sod/pot and Ammonium
oxalate in 2:3 ratio. Create calcium oxalate,
sod causes shrinkage, ammonium causes
swelling.
• Heparin: Directly inhibits coagulation by
binding to antithrombin III and leads to
conformational change in AT which inactivates
coagulation factors.
• Sodium Fluoride: Inhibits red cell glycolytic
enzyme pathway.
• Red vacutainer: Clot activator in the form of
silica. Yields serum.
• Different kinds of anticoagulants used.
• Mechanism of action of each
• 2-3 tests done for each vacutainer.
• Which anticoagulant is there in which
vacutainer.
Wintrobe Tube
Westergren Pipette
1. ESR full form
2. Principle
3. Phases of ESR:
• Phase of rouleaux formation: Initial 10 minutes
• Phase of settling: Next 40 minutes
• Phase of packing: Last 10 minutes
• Expressed as mm per first hour
4. Factors affecting ESR:
• Internal factors: Anemia, Polycythemia, Plasma fibrinogen levels
• External factors: Length of tube, bore of tube, position of tube
• Methods of ESR estimation:
• Westergren’s method
• Wintrobe’s method
• Micro ESR method
• Automated method
• Zones after 1 hour: Plasma, buffy coat, packed RBCs
• Plasma color can give additional information, buffy coat can be
used for parasites, LE cells
• Causes of elevated ESR: Anemia, increased immunoglobulins (TB,
MM, RA)
• Causes of low ESR: Polycythemia, sickle cell anemia, CHF,
Spherocytosis, hypofibrinogenemia
• Types of anti coagulants used for different ESR estimation methods
• Advantages of Wintrobe method
• Advantages of Westergren’s method
• Differences between Wintrobe’s tube and Westergren’s pipette
• Westergren’s: Pipette, longer, 30 cm length, 0-200 mm markings.
• Wintrobe’s: Tube, shorter, 11 cm long, two set of markings in
opposite direction from 0-10cm.
 Sahli’s Hemoglobinometer
• Components:
• Sahli’s Colour Comparator
• Sahli’s hemoglobin pipette
• Sahli’s Hemoglobin Tube
• Stirrer
• N/10 HCl
• Methods for Hb estimation.
• Reference method for Hb estimation recommended by
ICSH
• Disadvantages: uses Potassium cyanide in Drabkin’s
fluid, affected by Hyperlipidemia, high WBC count,
hyperbilirubinemia
• Method of Hb estimation in our lab: Electronic counter
• Principle of Sahli’s method
• Advantages and disadvantages of sahli method.
• Normal ranges of hemoglobin
• Anaemia.
Salah Bone Marrow aspiration needle

Trochar
Canula
Adjustable guard
• Parts of bone marrow aspiration needle.
• Sites of bone marrow aspiration in adults and children.
• Indications of BMA: Benign conditions: Refractory anaemia,
pancytopenia, PUO to look for granulomas, LD bodies, lipid storage
disorders
• Suspected leukemia, lymphoma (for staging), metastatic carcinomas
• BMA vs BMBx
• BMA is better for studying individual cell morphology
• BMBx: Better for studying overall cellularity, architecture, focal
lesions, packed marrow, myelofibrosis which yields dry tap
• Bone marrow biopsy needle name
• Bone marrow findings in Anemia, MM
P/S smear
• The parts of a peripheral smear
• Part used for doing DLC and RBC morphology
evaluation
• Stain used, the principle, names of romanowsky stains
• Information that can be gathered from p/s
• RBCs: Abnormal shapes and sizes
• WBCs: Numbers and percentages, abnormal cells, toxic
granules, Leukemias
• Platelets: Numbers
• Hemoparasites: Malaria, microfilaria etc.
Tissue Moulds
• Steps of histopathology
processing
• Stain used in histology
Coplin Jar
• Used for
transportation of
slides in alcohol
for pap stain
• Can be used for
performing steps
of staining as well
• Pap smear,
cervical cancer
Franzen Handle
• FNAC: Fine Needle Aspiration Cytology
• Procedure: Tissue aspirated with a small caliber needle,
spread on a slide, air dried and stained with giemsa.
• Wet smears fixed in alcohol and stained with pap stain.
• Uses: Used for superficially palpable lesions
• Deep lesions can be accessed via USG and CT
• Advantages: Minimally invasive, quick reliable results,
surgery can be avoided for benign or medically
treatable lesions
• Disadvantages: invasion of capsule cannot be
commented upon: FA vs FC
• Organs commonly sampled by FNA: Lymph node,
breast, salivary gland, subcutaneous swellings like
lipomas, Ecs
RBC pipette
RBC pipette WBC pipette
Red bead White bead
Red end White end
Markings till 101 Markings till 11
Hayem’s fluid, Dacie’s fluid used as diluent Turks fluid used as diluent in 1:20 ratio
in 1:200 ratio GAA, DW, GV
Hayem’s fluid uses Mercuric chloride Uses
Formaldehyde, sod. Citrate
Uses
Normal WBC count
Normal RBC count Normal differential values
Anemia Causes of leukocytosis, leukopenia
Urinometer

Stem

Float

Weight
• Use
• Specific gravity urine: weight of 1ml volume of
urine o weight of 1 ml of water
• Depends on various solutes in water.
• Normal range: 1.003 to 1.030
• Causes of low specific gravity: Tubular diseases,
high fluid intake, DM
• Low fixed specific gravity: DI, CKD
• High specific gravity: Dehydration
Neubauer’s Chamber
• What all can be counted on Neubauer’s chamber
• WBC count of blood and CSF, RBC count, platelet
count, eosinophil count, sperm count
• Diluting fluids:
• WBC: Turk’s fluid
• RBC: Hayem’s, Dacie’s fluid
• Platelets, sperm count:1% Ammonium oxalate
• Eosinophil: Dunger’s fluid