Basic Structure and Function Practical PDF

Basic structure and function BY Dr.Eman AbdelRaouf Lecturer of Medical Physiology  Hemoglobin content Estimation the amount of hemoglobin in 100ml blood. Method:  Sahli-Adam’s Hemometer (plastic frame with 2 standard colored glass, graduated tube & pi...

Basic structure and function BY Dr.Eman AbdelRaouf Lecturer of Medical Physiology  Hemoglobin content Estimation the amount of hemoglobin in 100ml blood. Method:  Sahli-Adam’s Hemometer (plastic frame with 2 standard colored glass, graduated tube & pipette).  0.1 HCl , dropper.  Alcohol, cotton &sterile lancets  Procedure:  Fill the graduated tube of Sahli Hemometer to mark 5 with 0.1 HCl.  Drawn blood from a thumb puncture into a pipette till mark 20 uL, then transfer it to the graduated tube and mix well until formation of a brown, yellow solution (HCL induces rupture of RBCs & react with Hb to form acid hematin)  After few minutes start to add distilled water drop by drop and mixing until the brown color in graduated tube matches the standard tube’s color.  Read the concentration of HB in gram / 100ml.  Normal value:  In adult male: 14 – 18 gm % (gm/100 ml blood).  In adult female:12-16 gm % (gm/100 ml blood). Male values is higher than that of female because male has androgen which is anabolic &↑ RBCs formation also has no blood loss as menses. Child: 11-14 gm % (gm/100 ml blood)  More in newborn (may reach 20 gm %) Due to relative hypoxia in intrauterine life. Hb content decreased in anemia & increased in polycythemia.  The method used for measurement of Hb in this picture is ………. The color seen in the tube shown in the picture is a result of formation of …………., due to the release of……. from ……….., following their hemolysis by ………... RBCs count: Number of RBCs / 1 mm3 Instruments: 1. Hemocytometer glass slide + slide cover  3 mm 3 mm 1 mm 1 mm  1 mm COVER Volume = 1 x 1 x 0.1 = 0.1 mm³ 0.1 mm height 1 mm 2. RBCs counting pipette 3-Light microscope 4-Diluting fluid: ISOTONIC Physiologic saline (0.9% NaCl), to avoid osmotic lysis of RBCs. Dilution is a method of anticoagulation in this test. Procedure Slide placement and microscope adjustment. Blood withdrawal: to the 0.5 mark. Diluting fluid aspiration: to the 101 mark. Mixing inside the bulb (by gentle rotation), Discard cell-free fluid, so the resulting dilution in the bulb is 1 to 200. Note the colors! Load the diluted blood onto the hemocytometer by touching the angle between the edge of the cover glass and the hemocytometer slide by an angle of 35°. So, the fluid will run under the cover. The cells are permitted to settle for several minutes. Counting method in 5 large squares (each with 16 small squares) Calculation of RBCs number in 1 mm 3 The number of RBCs in: First square = Second = Third = Fourth= Fifth = Total= Total number of cells counted X 10,000 =…../mm3 Why the counted number in 5 squares is multiplied by 10000 10000 = 50 x 200  50 Volume factor  200 dilution factor  Norma l value: Normally: - In newly born= 6-8 millions/mm3 -In children = 3.5 –5 million/mm3 -In adult male = 4.5-6.5 million/mm3 -Female = 3.8-5.8 million/mm3 -RBCs count Increases in hypoxia &polycythemia. - RBCs count decreases in anemia. Male value > female ??? Newborn > adult ??? Identify the equipments shown in the pictures above. They are used in the estimation of ………………….  RBCs volume: (Hematocrit value) (packed cell volume) It is the volume of RBCs contained in 100 ml of blood (the blood must be fully oxygenated, unhemolysed &uncoagulated) Equipment:  Microhematocrit tube (75 x 1 mm & heparinized).  Microhematocrit centrifuge.  Microhematocrit tube reader.  Sterile lancet, 70 % ethyl alcohol. Micro-hematocrit tube  Micro-hematocrit Micro-hematocrit tube reader centrifuge  Procedure: Obtain blood drop by pricking the thumb. Fill the micro-hematocrit tube up to it's 2/3 with blood sample by touch the drop of blood by one end of the tube. Close the empty end of the tube by a piece of plasticine. Centrifuge the tube at 12000 per minute for 5 minutes. Remove the tube and read Ht V. by putting the tube on special micro hematocrit scale. Micro-hematocrit tube reader Normal Values Male= 47% (42-54%) Female= 42% (37-47%) Children= 40% (36-44%) Newborn= 50-58% Pathological changes  Increases by:-increased RBCs (polycythemia) -decreased plasma (dehydration or hemo- concentration as after burn) - Decreases by:-decreased RBCs (anemia) -increased plasma (overhydration) - Used in calculation of blood volume, Renal blood flow and blood indices  Physiologically Ht. value is more: In venous blood than in arterial blood as RBCs volume is large in venous blood due to chloride shift phenomenon. In large vessels due to skimming phenomenon as RBCs prefer to pass in large vessels than in small blood vessels Chloride shift phenomenon Skimming phenomenon Male values(Hb content, RBCs count, Ht value) is higher than that of female because male has androgen which is anabolic &↑ RBCs formation has no blood loss as menses. More in newborn due to relative hypoxia in intrauterine life. Blood indices These are groups of laboratory findings which are done for diagnosis of different types of anemia. Blood indices Mean corpuscular volume (MCV) Mean corpuscular hemoglobin concentration (MCHC) Calculate the blood indices from the given Hb content, RBCs count and Hematocrit value: Hb = 15 gm% RBCs count = 5 million/mm3 Ht =45% Thank you