White Blood Cell PDF

12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula Immunology Book (Https://Labpedia.Net/Category/Elementary-Immunology/) Lab Tests...

12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula Immunology Book (Https://Labpedia.Net/Category/Elementary-Immunology/) Lab Tests ‣ Lectures Blog (https://labpedia.net/) Medical Dictionary (Https://Labpedia.Net/Medical-Dictionary/) ‣ ‣ About Us (Https://Labpedia.Net/About-Us/) Contact (Https://Labpedia.Net/Contact/)  White blood cell:- Part 2 – Total Leukocytes Count Procedure, TLC Solution Preparation Hematology (https://labpedia.net/category/lab-tests/hematology/) Lab Tests (https://labpedia.net/category/lab-tests/) Table of Contents (https://labpedia.net/white-blood-cell-part-2-total-leukocytes-count/#) 0.0.1. White blood cell(https://labpedia.net/white-blood-cell-part-2-total-leukocytes-count/#White_blood_cell) 1. Total leucocytes count (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- procedure (TLC) count/#Total_leucocytes_count_procedure_TLC) 1.0.0.1. Sample(https://labpedia.net/white-blood-cell-part-2-total-leukocytes-count/#Sample) 1.0.0.2. Purpose of the test (Indications) for (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- Total leucocytes count count/#Purpose_of_the_test_Indications_for_Total_leucocytes_count) 1.0.1. Principle of white cell count (total (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- leucocytes count) count/#Principle_of_white_cell_count_total_leucocytes_count) 1.0.2. Pathophysiology of Total (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- leucocytes count TLC) count/#Pathophysiology_of_Total_leucocytes_count_TLC) 1.0.3. White blood cells are (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- divided into: count/#White_blood_cells_are_divided_into) 1.0.4. Preparation of Total leucocytes (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- count (TLC) solution: count/#Preparation_of_Total_leucocytes_count_TLC_solution) 1.0.4.1. Total leucocyte count (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- Calculations: count/#Total_leucocyte_count_Calculations) 1.0.4.2. Normal values of total (https://labpedia.net/white-blood-cell-part-2-total-leukocytes- leucocytes are: count/#Normal_values_of_total_leucocytes_are) White Blood Cell (Https://Labpedia.Net/White-Blood- Cell%E2%80%A6leukocytes-Count/%20%E2%80%8E) Total Leucocytes Count Procedure (TLC) Sample 1. EDTA blood is needed. 2. Oxalate or citrated blood can also be used. Purpose Of The Test (Indications) For Total Leucocytes Count 1. To differentiate between acute and chronic infection.  2. To follow the patient with chemotherapy. 3. To find the effect of drugs. file:///C:/Users/Babina Aryal/Downloads/White blood cell (Total leucocyte) count and TLC solution formula.mhtml 1/22 12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula Principle Of White Cell Count (Total Leucocytes Count) Blood is diluted with a solution (acetic acid) that causes the lysis of RBCs. Acetic acid has no effect on the WBCs. Gentian violet is added to differentiate the WBCs. Pathophysiology Of Total Leucocytes Count TLC) 1. White blood cells develop from the stem cells in the bone marrow. 2. Stem cells differentiate into: 1. Granulocytic series cells. The granulocytes got the name due to the presence of distinct granules in the cytoplasm of these cells. 1. The life span of leucocytes varies from 13 to 20 days; after that, these cells are destroyed in the lymphatic system and may be excreted from the body in feces. 2. Leucocytes fight infections and defend the body by a process called phagocytosis. 3. WBC serves as an excellent indicator of the various disease process. 2. Non-Granulocytic cells. 3. The granulocytic series consists of: 1. polymorphonuclear leucocytes (Neutrophils). 2. Eosinophils. 3. Basophils. 4. Monocytes.  file:///C:/Users/Babina Aryal/Downloads/White blood cell (Total leucocyte) count and TLC solution formula.mhtml 2/22 12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula (https://www.labpedia.net/wp-content/uploads/2020/01/WBC-development-new.jpg) Development of white blood cells White Blood Cells Are Divided Into: (Https://Labpedia.Net/White-Blood- Cell%E2%80%A6leukocytes-Count/%20%E2%80%8E) 1. Phagocytic cells: 1. Polys. 2. Eosinophils. 3. Basophils. 4. Monocytes. 2. Immunocytic cells: 1. B- lymphocytes. 2. T-lymphocytes are: 1. T-helper cells (CD4+) 2. T-cytotoxic cells (CD+) 3. T-effector cells (CD4+)  file:///C:/Users/Babina Aryal/Downloads/White blood cell (Total leucocyte) count and TLC solution formula.mhtml 3/22 12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula Preparation Of Total Leucocytes Count (TLC) Solution: 1. Principle of the TLC method: 1. Blood is diluted with a fluid that causes the RBCs’ hemolysis, but WBCs remain intact, and then these are counted in the Neubauer chamber. 2. Gentian violet lightly stains the leucocytes and allows those to be counted. 2. Chemicals (Reagents) needed are: 1. Glacial acetic acid = 2 ml. 2. Gentian violet (1%) = 1 ml. 3. Distal water = 97 ml. 4. This is basically a 2% solution of acetic acid. 1. Add 2 ml of glacial acid (2 + 1), 1 mL gentian violet (1 %), and add 97 ml of distle Water to make up to 100 ml solution. 2. Gentian violet is added until the color is pale blue-violet. 3. Total leucocyte count or WBC pipette: 1. This pipette (also called Thoma pipette) long stem is divided into two parts: 1. The long stem is marked with 0.5 and 1.0 2. While the short arm after the bulb is marked 11. 3. Its central portion is a bulb or a globular shape with one white bead in it. 4. Rubber tubing is attached to suck the blood. 5. Ultimately the dilution of the blood to the TLC fluid is 1:20. (https://www.labpedia.net/wp-content/uploads/2020/01/wbc-pipette-improved-3.jpg) WBC pipette Total leucocytes count Procedure: (https://labpedia.net/white-blood-cell%E2%80%A6leukocytes- count/%20%E2%80%8E) 1. Take the TLC pipette, which has a white bead inside. 2. Total leucocytes count (WBC) pipette method: 3. Fill the blood into the 0.5 marks and then add the TLC solution. 1. Fill the pipette with the TLC solution to point 11. 2. Remove the rubber tubing. 3. Seal both ends or hold in between two fingers. 4. OR can put this pipette on the mechanical device to shake it. 5. Shake for 1 minute or preferably for 2 minutes. 6. Shaking is important before filling the Neubauer chamber. 7. After thorough mixing, discard the first few drops and then gently fill the chamber until the platform is filled.  8. The capillary action will draw the fluid. file:///C:/Users/Babina Aryal/Downloads/White blood cell (Total leucocyte) count and TLC solution formula.mhtml 4/22 12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula 9. Allow the chamber on the microscope stage for 2 to 3 minutes till the cells are settled. 4. Tube method: 1. Take 0.02 mL blood and mix it with diluting fluid. 2. Take TLC dilution fluid 0.38 mL in a small tube and mix it with the blood. (1:20 dilution). 3. Mix them very well. 4. The dilution will be the same. 5. This tube method is more accurate than the Thoma pipette technique. (https://www.labpedia.net/wp-content/uploads/2020/01/WBC-count-tube-method.jpg) white blood cell – WBC count tube method 5. In case of low WBC count, fill the pipette to mark 1, and this will give dilution of 1:10, OR 1. Take 0.1 mL blood and 0.9 mL diluent solution (1:10 dilution). 2. In the case of a high WBC count, then make higher dilution.  file:///C:/Users/Babina Aryal/Downloads/White blood cell (Total leucocyte) count and TLC solution formula.mhtml 5/22 12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula (https://www.labpedia.net/wp-content/uploads/2020/01/neubar-chamber-photo-1.jpg) Neubauer chamber (https://www.labpedia.net/wp-content/uploads/2020/01/WBC-TLC-counting.jpg)  Neubauer chamber counting of white blood cells (TLC) file:///C:/Users/Babina Aryal/Downloads/White blood cell (Total leucocyte) count and TLC solution formula.mhtml 6/22 12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula (https://www.labpedia.net/wp-content/uploads/2020/01/neubar-chamber-filling-by-wbcs-solution-1.jpg) Procedure to fill the Neubauer chamber  file:///C:/Users/Babina Aryal/Downloads/White blood cell (Total leucocyte) count and TLC solution formula.mhtml 7/22 12/2/24, 12:26 PM White blood cell (Total leucocyte) count and TLC solution formula (https://www.labpedia.net/wp-content/uploads/2020/01/WBC-sample-of-one-square-1.jpg) WBCs counted in one of the large squares as a sample Total Leucocyte Count Calculations: 1. Count the cells in the Neubauer chamber. These are counted in the four large corner squares labeled as WBC and if the number is Y. 1. One large area is 1 x 1 mm, and the depth is 0.1 mm. 2. Total area counted in 4 large squares = 4 x 1 x o.1 = 0.4 µL (4/10). 3. Y x 10/4 is the total WBC in the cell in 1 µL. 4. Now dilution is 1:20. 5. Number of WBC in 1µL = Y x 10 x 20/4 = Y x 50 = Total WBC count. 6. Total TLC = counted cells (Y) x 50 = TLC/cmm. 2. If the count is low