Separation of Plasma and Serum from Whole Blood
WHOLE BLOOD :
- The average person circulates about 5L of blood (1/13 of body weight), of which 3L is plasma and 2L is cells.
- It is living tissue, composed of different types of cells suspended in fluid called plasma.
- The cells are produced primarily by bone marrow and account for blood “solids”.
- Plasma fluid derives from the intestines and organs and provides a vehicle for cell measurement.
Radial Immunodiffusion
BLOOD FUNCTIONS :
- Transportation of gases, nutrients, waste products, regulatory molecules(e.g. hormones) and metabolites.
- Regulation of pH and osmotic pressure.
- Maintenance of body temperature.
- Protection against infections.
- Clot formation,
BLOOD CELLS :
-
- Blood cells are classified as
- Red blood cells (erythrocytes)
- White blood cells (leukocytes)
- Platelets (thrombocytes)
- The size of the cells differs:
- white cells are the largest, red cells fall into the middle, and platelets are the smallest.
RED BLOOD CELLS (RBC) :
- Red blood cells contain hemoglobin, a complex iron-containing protein that carries oxygen throughout the body and gives blood its red color.
- They live for approximately 120 days in the circulatory system and are eventually removed by the spleen.
WHITE BLOOD CELLS (WBC) :
- They are responsible for protecting the body from invasion by foreign substances such as bacteria, fungi, and viruses.
- Types of WBCs:
- Granulocytes
- Neutrophils
- Eosinophils
- Basophils
- Agranulocytes
- Monocytes
- Lymphocytes
PLATELETS :
- They are very small cellular components of blood that help the clotting process by sticking to the lining of blood vessels.
- They survive in the circulatory system for an average of 9-10 days before being removed from the body by the spleen.
- They help prevent massive blood loss resulting from trauma, as well as blood vessel leakage.
PLASMA :
- Plasma is the liquid portion of the blood
- It constitutes about 55 % of blood volume.
- 90% of plasma is water
- It contains:
- Albumin (the chief protein constituent),
- Fibrinogen (responsible, in part, for the clotting of blood),
- Globulins (including antibodies).
SYNTHETIC CULTURE MEDIA
SERUM :
- It resembles plasma in composition but lacks the coagulation factors. (Serum = Plasma – clotting factors)
- It is obtained by
- letting a blood specimen clot prior to centrifugation usually in a red top tube with no additives or anticoagulant.
- Or by centrifugation of plasma to precipitate fibrinogen.
- The serum is preferred for many tests (e.g. determination of lactate dehydrogenase) as the anticoagulants in plasma can sometimes interfere with the results.
COLLECTION OF BLOOD SPECIMENS :
-
- If whole blood or plasma is desired, an anticoagulant must be added to the specimen immediately after it is drawn or placed into the tube into which the blood is collected.
- Types of Anticoagulants
Heparin :
- It is the most satisfactory anticoagulant since it does not produce a change in red cell volume or interfere with subsequent determinations.
- It inhibits the formation of thrombin from prothrombin and thus preventing the formation of fibrin from fibrinogen.
EDTA :
- It is a chelating agent, drives its anticoagulant activity from the fact that it binds calcium, which is essential for the clotting mechanism.
Potassium Oxalate :
- It inhibits blood coagulation by forming insoluble complexes with calcium ions, which is necessary for coagulation.
Sodium Citrate :
- It does not precipitate the calcium but converts it into a non-ionized form, and hence prevent clotting of blood.
Sodium Fluoride ;
- It acts as a weak anticoagulant.
- It has been used chiefly as a preservative since it inhibits red cell metabolism and bacterial action. If blood is treated to prevent clotting and permitted to stand or centrifuged in a container:
- The RBCs, which weigh more than the other components, will settle to the bottom;
- The plasma will stay on top, and •
- The WBCs and platelets will remain suspended between the plasma and the RBCs.
CHANGES IN BLOOD ON KEEPING :
- Loss of carbon dioxide.
- Conversion of glucose to lactic acid (glycolysis).
- Increase in plasma inorganic phosphate.
- Formation of ammonia from nitrogenous substances.
- Passage of substances through the red cell envelope.
- Conversion of pyruvate into lactate.
MATERIALS :
- Whole blood
- Centrifuge (up to 5000 rpm)
- Centrifuge tubes suitable for the rotor of the centrifuge (preferably plastic and capped).
- Disposable gloves
- Disposable Pasteur pipette.
- Measuring cylinder 10 ml.
Translation In Prokaryotes And Eukaryotes
METHOD :
- Into dry clean centrifuge tube, pipette 15 ml of whole blood (V1).
- Place the centrifuge tube in the centrifuge machine and run it at 3000 rpm for 10 minutes. Centrifugation of whole blood separates the solid from the supernatant plasma.
- Remove the tube, withdraw the liquid layer (plasma) by pasture pipette and measure its volume using a small measuring cylinder (V2). Determine the volume of blood cells too V3 (equal to V1 – V2).
- Transfer the supernatant (plasma) in another centrifuge tube and make further centrifugation at 3000 rpm. This will precipitate fibrinogen and the supernatant will be SERUM. Measure its Volume (V4).
RESULT :
Record your results in the following table:
😂😊✌❤🤞Download Separation of Plasma and Serum from Whole Blood E-Book Pdf Form Clock Here,..❤🤞�✌🤞✌🤞
😊😊😍❤❤❤✌✌😊😊😍😍
Lehninger Principles of Biochemistry 7th Edition Pdf Free
what is serum, Separation of Plasma and Serum from Whole Blood, fibrinogen, thrombin, plasma, and serum difference