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Blood. Blood-based tumor biomarker discovery - Bioanalysis Zone.

FUNCTIONS. Transport Respiratory Prevention of hemorrhage Defense Regulatory.

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Blood Composition.

VL,NSV1d gos.

1. Red Blood Cells (RBCs or Erythrocytes) 2. White Blood Cells (WBCs or Leucocytes) 3. Platelets (Thrombocytes).

Carrier of O 2 and CO 2 5 * 10 6 /mm 3 Polycythemia Anemia.

90 % Water. 10 % solid. Plasma. Non Diffusible. High molecular weight cannot pass the walls of blood vessels e.g. Albumin, Globulin, Fibrinogen Enzymes Nucleoproteins Hormones.

O 2. N 2. Gases. CO 2. Dissolved in plasma.

Arterial Blood is Red ( oxyhemoglobin ) Venous Blood is Bluish (reduced hemoglobin).

the osmotic pressure of Blood is equivalent to 0.9 % NaCl colloidal O.P. (due to plasma proteins) is more effective in water diffusion across capillary walls than Crystalloid O.P. (mainly due to NaCl ).

Collection of Blood Specimens. Capillary Blood Collection.

Venous Blood Collection.

After a pulse is found, a blood sample is taken from the artery.

Tools. Disposable. Dry. Precautions in Blood Sampling.

After Blood Collection, it’s essential to avoid Hemolysis, especially if serum or plasma is to be used. If hemolysis is not noticeable on inspection, it can be detected spectroscopically by the presence of oxyhemoglobin ..

Glucose Lactic acid. Sodium Fluoride used to inhibit glycolysis.

It is the conversion of blood from a free flowing liquid SEMISOLID GEL.

Clot formation:. Ca 2 +. Platelets.

Ca 2+. Platelets. Thrombin. Fibrinogen. Prothrombin.

A human blood sample showing red cells (erythrocytes), white cells (leukocytes), platelets (colored yellow/green) and fibrin (light brown). The image was captured by field emission scanning electron microscope and then was colored. https://www.facebook.com/NeuronsWantFood.

1) Heparin ( antithrombin ) Prothrombin thrombin 2 ) Dicoumarol ( oral anticoagulant) Prevent synthesis of prothrombin due to structural similarity with vitamin K 3) Ethylene diamine tetraacetic acid chelate Ca ++ 4) Oxalate precipitate Ca ++ 5) Sodium citrate convert Ca ++ to non-ionized form 6) Sodium fluoride Act as anticoagulant but large amount are required. It is used mainly as preservative (by inhibition of RBCs metabolism and bacterial action ).

Blood (anticoagulant) Water bath 37 Centrifuge. Blood + anticoagulant Centrifuge.

Differences between Plasma and Serum. Plasma Serum Preparation Blood + anticoagulant then centrifuge Blood without anticoagulant then centrifuge Thrombin Absent Present Injection Can be injected Never injected Fibrinogen Present Absent Prothrombin Present Absent Addition of xss Ca ++ Clotting No clotting.

Carboxyhemoglobin Hemoglobin Lactate pH Pyruvate.

Hematocrit Value (HV) Or Packed Cell Volume (PCV).

Wintrobe tube.

Dehydration. Normal HV values 45% for adult male 35% for adult female 25% for children 65% for new born.

Erythrocyte Sedimentation Rate ( ESR ). Experiments on Blood.

Westergren tube. One hour later C 2009 American colle of Rheumat(.

Physiologically. Normal ESR Male up to 19 mm/ hr Female up to 15 mm/ hr Children up to 13 mm/ hr.

Fragility test. It’s the resistance of RBCs to hypotonic solutions In normal human, complete hemolysis occurs only below about 0.35% NaCl . i.e. RBCs resist hemolysis above 0.45%.

Hypertonic ooo Isotonic Hypotonic.

crenated HYPERTONIC normal swollen lysed VERY ISOTONIC HYPOTONIC HYPOTONIC.

Prepare a series of hypotonic NaCl solutions. 0.3% NaCl 0.4% NaCl 0.45% NaCl 0.5% NaCl 0.6% NaCl 0.7% NaCl 3 ml 4 ml 4.5 ml 5 ml 6 ml 7 ml 1% NaCl solution 7 ml 6 ml 5.5 ml 5 ml 4 ml 3 ml Distilled water.

Transparent tubes indicates Hemolysis Translucent tube indicates Partial Hemolysis.

Effect of Different Hemolytic Agents. 0.1 N NaOH.

Formation of brown Alkaline Hematin. Formation of brown Acid Hematin.

Determination of hemoglobin. Determine the oxygen carrying capacity of blood.

Acid Hematin Method ( Sahli ’ s method ). Blood. dil. Acid.

Sahli ’ s Hemometer. Sahli Type Haemoglobin Meter, Digital Hemoglobinometer, Hemoglobin Test Machine, Hemoglobin Monitor, Hb Meter, Hemoglobin Check Machine in Industrial Estate, Ambala , Laby Instruments Industry | ID: 15672891530.

Colorimetric method ( Drabkin ’ s reagent). Drabkin ’ s reagent.

Sahli ’ s method Drabkin ’ s method Visual method Colorimetric method Subjective Objective Less accurate More accurate 0.1 N HCl ( reagent ) Drabkin ’ s reagent Acid hematin (product ) Cyano Met Hb (product ).

Normal Hemoglobin values Female : 12-16 g/dl (70-110%) Male : 14-18 g/dl (80-120 %).

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