Materials for auditory self-work.
4.1. List of study practical tasks necessary to perform at the practical class.
It’s necessary for work: microscope, counter of Goryaev, scarificators, mixers, solutions, alcohol, cotton wool, photoelectrocalorymeter (PEC), apparatus for erythrocytes automatic count, hemometer of Sali, hydrochloric acid solution, alkaline solution, device for hemolysis assessment.
Task 1. To determine erythrocytes amount in blood.
In some clinics, unfortunately, this old method is still widely-used. Under modern conditions this method was changed on automatical one. Automatical devices have special instructions on their expluatation.
Erythrocytes number estimation in Goryaev’s chamber.
To petch investigated blood in special mixer (melanger) till the mark 0,5 or 1,0 (it depends on blood dissolving). To wipe mixer end with cotton wool and to petch 3% NaCl in it till the mark “101”. To mix carefully mixer content in course of 1 minute, to pour 1-2 blood drops on cotton wool and to fill up Goryaev’s chamber with the next blood drop. But one should grind covering glass to Goryaev’s chamber before this. Erythrocytes are estimated in 5 large squares (each of them is divided into 16 small squares) placed diagonally (obliquelly) to the net. It’s necessary to estimate red blood cells located inside every small square as well as on its superior and left boundaries. One should put found erythrocytes amount under following formula:
| X= (a x 4000 x 200 or 100) : 80 x 106, |
where:
X – erythrocytes amount,
a - erythrocytes amount in 5 large (80 small) squares.
1/4000 mcl/mm3 – one small square volume;
200 or 100 – blood dilution degree;
106 – co-efficient for re-computation into SI.
Task 2. Hemoglobin content determining in blood.
Like in a case with erythrocytic amount, there are both rutine (old) measurements methods and new, automatical ones (on PEC, hemoglobinometers and others).
Hemoglobin content determination by Sali method.
To pour 0,1 normal (approximately 0,2 ml) hydrochloric acid solution in graduated pipette of Sali till inferior ring level. To petch exactly 0,02 ml of blood with pipette from hemometer and to blow it on the test tube floor. Shaking up the test tube, to mix its content carefully. The mixture must stay in course of 5-10 minutes at room temperature. Hydrochloric acid causes erythrocytic hemolysis and hemoglobin destruction. Releasing hem interacts with hydrochloric acid and is transformed into hydrochloric hematin. Test tube content becomes dark brown as a result of this reaction. The investigator must add distillate water in 5-10 min till investigated liquid color becomes equal to the standard solution color. To mark on the scale at which level hydrochloric hemolytic is. Received ziphra multiple on 10. Result (product) will correspond to hemoglobin concentration in investigated blood in g/l.
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Task 3. To estimate blood color index.
Color index characterizes erythrocytes satiation degree with hemoglobin. It is calculated on formula:
C.I.= (X hemoglob. x 5,0 x 1012/l) : (167 g/l x X erythroc.), where:
X hemoglob. – found hemoglobin amount (g/l);
X erythroc.- found erythrocytes amount in 1 l of blood.
The second formula: Hb (g/l) x 3: RBC (3 first ziphras). It is evaluated in conditional units.
Normal values:
· 0,75-1,0 – erythrocytic normochromy;
· more than 1,0 – hyperchromy;
· less than 0,75 – hypochromy.
Literature recommended:
1. Lecture course.
2. Mistchenko V.P., Tkachenko E.V. Methodical instructions for medical students (short lecture course).-Poltava, 2005.-P. 62-65.
3. Mistchenko V.P., Tkachenko E.V. Blood system Physiology //Methodical recommendations to practical classes for students of medical and dental departments.-Poltava, 2005.-20p.
4. Kapit W., Macey R.I., Meisami E. The Physiology Colouring Book: Harpers Collins Publishers, 1987.-P. 136.
5. Guyton – Ganong – Chatterjee. Concise Physiology /Ed. By Dr Raja Shahzad Gull: M.B.B.S., F.C.P.S., King Edward Medical College.-Lahore, 1998 (1st Edition).-P.171-178, 204.
6. Stuart Ira Fox. Human Physiology.-8th Ed.-McGrawHill, 2004.-P.368-369, 371-372.
7. Seeley R.R., Stephens T.D., Tate P. Essentials of Anatomy and Physiology.-The 3rd Ed.-McGraw Hill, 1999.-P.288-291, 299.
Materials for self-control:
Control questions:
1. Erythrocytes structure and quantity, their amount changings under physiological conditions.
2. Erythrocytes functions.
3. Erythropoiesis regulation, specific and non-specific erythropoiesis regulative ways. Salivary glands role in this process.
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4. Hemoglobin molecule structure, hemoglobins types.
5. Hemoglobin functions.
6. Hemoglobin chemicals in blood.
7. Color index.
8. Erythrocytic hemolysis, its types.
9. Hemolysins.
10. Different environments and solutions influence on erythrocytic hemolysis.
LESSON 33
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