how to count rbc in neubauer chamber

Nowadays, two types of RBC Diluting fluid are commonly used in Laboratories . Do you know about the functionality of the hemocytometer? ,8Jl?IT (@Lx,MB @"F],c#!Y1OPN 3Cm/km~aO, T^{e~yq $AvN% D Note: This cover glass is 0.4mm thick and has a smooth surface. Can you plz explain from your example total wbc count is multiplied by 50 how this fifty comes??? Platelet Counting AreaThe large center square is used to count platelets. Using the 10X objective, focus both onto the grid pattern and the cell particles. The diluted blood is placed in a counting chamber and the cells are counted under the microscope. Take 3.98 ml RBC diluting liquid in a clean, dry and grease-free Test tube. Use a flat surface to place the chamber, like a table or a workbench. Mix for a few minutes, and you are ready to use your Hemocytometer/Neubauers Chamber. EDTA serves as an anticoagulant or chelating agent, which prevents blood coagulation by binding with the calcium ions. RBC-Diluting Fluid . Each square of the Central Square, divided into 25 squares, contains 16 squares. The blood cells are then counted in the volume of diluted blood in Neubauers counting chamber or Haemocytometer. The counting can be done either in the central large square or in the corner squares, depending on the size of the cells under study. How much does a smile makeover in India typically cost? This test is usually performed during a complete blood count (which also measures other types of cells). the motile promastigotes may . It is also known as Red Blood Cell. The Blood Specimen contains a large number of Red Blood Cells. of Red Blood Cells to less than 3.5 million/mm3). 4HVCP5VXo1MRw xlnUD+ (e:HaRmcPu:PK|Lx,S`P+z`xa+-HZ[HNJFhf#GiMf_ a;iJ7F8Q/C. Enjoy notes in your phone without ads. 2021 Medical Laboratory Scientist - MLS All Rights Reserved. This technique is used to measure RBC in blood and body cavity fluids (peritoneal and pleural). mm, The RBCs can be counted in five squares out of these 25 squares. Thanks The coverslip is a square glass of thickness 22 mm. The consent submitted will only be used for data processing originating from this website. Remove the Neubauers chamber/Hemocytometer from its box and clean it with a cotton swab or gauze. %PDF-1.5 The above Composition is based on HIMEDIA Hayems diluting fluid protocol which you can check here. Neubauer chamber's counting grid is 3 mm x 3 mm in size. Red blood cells are first sphered in a diluent before being passed through a laser detector using the hematology analyzer at Cornell Universitys Clinical Pathology Laboratory. For this, the blood specimen is diluted (usually in 1:200 ratio) with the help of RBC diluting fluid (commonly the Hayems Fluid) which preserve and fix the Red blood cells. The diagram below provides a well-labelled presentation of an RBC pipette. Allow for 2 minutes to settle the cells. Want to be notified when our article is published? MLT Lectures by Tanveer Tara 10.5K subscribers Subscribe 2.5K views 2 years ago Lab Practical's | Diagnosis | Biosafety As 10X is. (adsbygoogle = window.adsbygoogle || []).push({}); Paramedics World was founded in 2017 by Sahil Batra, with the sake to provide medical and paramedical information and reference meterial to the students and the professionals. INTRODUCING THE SPECIMEN INTO THE HEMOCYTOMETER/ NEUBAUER CHAMBER: By the help of micro-pipette, cautiously draw/pull up about 20ml of the cell dilution/mixture. Place the Neubauer chamber on the microscope stage; 10. We and our partners use cookies to Store and/or access information on a device. The formula for RBCs count is: It has two markings at the bottom as 0.5 and 1 and the top of the pipette is marked 101. To count the RBCs, you can perform microdilution and macrodilution quantitative methods by using Neubauer's chamber. 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The Capillary action allows a small amount fluid to be poured into the chamber using the pipette. This helps transport oxygen from the lungs into tissues and carbon dioxide back to the lungs for exhalation. The purpose of performing a total Red Blood Cell (RBC) count is to measure the number of red blood cells in a given blood volume. Gently press down on the rubber tube of your RBC pipette until the fluid drops are in hanging position. The RBC count is the total number of red blood cells contained in a sample of your blood, usually expressed as million cells per microliter (million cells/microliter). Accurately measure the amount of specimen and Diluting Fluid to avoid any error in the results. The glass cover is a squared glass of width 22 mm. So the Area of 5 small squares is. It has a 20mm width and is square-shaped. Depending on the counter, this technique uses electrical resistance changes to count cells and give an assessment of their volume. Each square has an area of 1/25 mm-squared (that is, 0.04 mm-squared) and depth of 0.1 mm. RBC Manual Count through Neubauer chamber and its calculation . There is usually a 0.1 mm gap between the glass cover and the central area of the haemocytometer. Laboratories use Macrodilution instead of Microdilution. Brochure Precision Scientific Instruments Corporation Darya Ganj, Delhi The formula to calculate the number of RBCs will be explained. Neubauer's Chamber or Haemocytometer; 2. So that when a cover slip is kept on the counting region, there is a gap of 0.1 mm (1/10mm) between the cover slip and the ruled area. Many countries ban the use of mouth pipettes (RBC pipette). Wipe off the first drop of blood. PPT NEUBAUER CHAMBER (Haemacytometer) PowerPoint Presentation ID from www.slideserve.com. <>/ExtGState<>/XObject<>/ProcSet[/PDF/Text/ImageB/ImageC/ImageI] >>/MediaBox[ 0 0 595.32 841.92] /Contents 4 0 R/Group<>/Tabs/S/StructParents 0>> He mentioned Nageotte calculates WBC in 50 uL, however, Neubauer counts WBC only in 0.1 uL x 5. Typically, the concentration range for a cell count with Neubauer chamber is between 250,000 cells / ml and 2.5 million cells / ml. Lets consider it as N no. Neubauers Chamber has ruled over the total area of 9 square mm. Cell counting areas in neubauer chamber: The counting can be done either in the central large square or in the corner squares, depending on the size of the cells under study. Red blood cell (RBC) count PROCEDURE 1. There are some diff. CELL COUNTING AND MICROSCOPIC FOCUSING ADJUSTMNT: Total no. Micropipettes are used in research and practical labs to disperse liquid of desired volume. Every of these 25 squares is additionally distributed into 16 small squares having single lines and every of these smallest squares having area around of 1/400 mm2. The big center square is used to count platelets. Capillaries are the smallest blood vessels located near the skins surface. It is a calculated value, not one that is directly measured. The deepness used in the formula is permanently remain 0.1. Before commencing work, thoroughly spray the inside of the laminar flow safety cabinet with disinfectant and wipe clean with tissue. Subsequently their concentration is less than RBCs, a big area is necessary to complete the count. CSF is dispersed directly into the counting chamber. EDTA is an anticoagulant, or chelating agent. However, Hayems diluting fluid gives the better results. Nowadays, most commonly Improved Neubauers Chamber is used and in some laboratories, other types of chambers are also employed like Burkers chamber, Levys chamber and Fusch Rosenthal chamber etc. 1/5. of cells to be counted must be used. Our website services, content, and products are for informational purposes only. Types, Causes of Jaundice, Molecular Biology Multiple Choice Questions: DNA Replication, Microbiology MCQs: Morphology of Bacteria, Pregnancy Test: Principle, Procedure, Requirements and Interpretation, Staphylococcus Aureus Cultural Characteristics, Place/Put the chamber under the microscope. Example calculations for WBC and RBC If you counted white cells The instrument detects and amplifies the pulse from each cell as it passes through the aperture. Use a smooth surface to place the chamber, like a board or a worktop. It is also known as Red Blood Cell Count t in Neubauer Chamber. Take 3.98 ml of RBC diluting fluid plus 0.02 ml of blood specimen (capillary or anti-coagulated blood) into a clean, dry test tube. Use the following formula to calculate the Total Red Blood Cell Count. some should pls help. It can be cumbersome in busy laboratories. Take a clean, grease-free haemocytometer slide and cover glass. endobj Modern devices, such as photometric and electrometric counters, can be more precise or automated. James M. Ramsey performed an experiment to measure how sampling area and dilution factors affected variation in cell counts. It is a square-shaped coverslip, having a width of 20 mm. To view the purposes they believe they have legitimate interest for, or to object to this data processing use the vendor list link below. It uses a micropipette to mix the blood specimen with the RBC diluting fluid. General features of the Neubauer's chamber. Then, you should carry the RBC pipette at an angle (45 degrees) and load a small volume of RBC sample towards the edge of a cover glass. After charging, wait for 3-5 min so that the cells settle down in the chamber & then focus the chamber under the microscope to calculate Red Cells. Let the mean be 'n'. Below is a diagram that shows the components and parts of a micropipette. It is a device invented by Dr Heinrich Schnitger. The counting area contains 2 square shaped ruled/lined zones. Wait for the cells to settle in the chamber. Charge the Hemocytometer/Neubauers chamber carefully. Remove the Neubauers chamber/Hemocytometer from the case and clean it with a cotton swab or gauze. Now, take out the RBC pipette and fill it with the Diluted Specimen, mix the solution well and then discard 1-2 drops from the pipette before charging the chamber. We can count Total Leucocytes Count or TLC in 10 x objective. All the . A volume of 10l is sufficient to fill out the one counting chamber. Counting yeast with a hemocytometer Hemocytometer calculator To count the RBCs, you can perform microdilution and macrodilution quantitative methods by using Neubauers chamber. RBC counts the cells in the 5 corners of the Central Square as well as the 4 Corner squares (divided to 25 squares) or 1 central square of Larger Central Square (divided to 25 squares). Modern devices, such as photometric and electrometric counters, can be more precise or automated. number of cells. The total number of cells per microliter of sample can be calculated from the number of cell counted and area counted. Microscope Lancet Different areas are used for counting red blood cells and white blood cells. The requirements for the RBC count method are as follows: This special type of chamber is used to count cells, particularly Blood cells. xZms6~:M He2Iq>@$*$.Es7l.gzYlU{_ly7W}~){Xz]nk)zi(i]oA{./qOmr./n|owh VJdZ))BH1M aDdj^{U.B, The amplitude is proportional to the cell size. The chamber has three parts. RED CELL COUNT The adjacent figure shows the rulings (improved Neubauer) which are inscribed on the counting chamber. Now gently place the pipette tip against the edge of the cover glass and if required slowly expel the more liquid until the counting chamber is full. Stop when the sample touches the three sides of the chamber (about 10 ml); do not overflow into the moat which surrounds the chambers. THE AIM / PURPOSE OF PERFORMING TOTAL RBC COUNT, PRINCIPLE OF TOTAL RBC COUNT USING HEMOCYTOMETER, The composition of Hayems diluting Fluid, The composition of Formalin Citrate diluting fluid, Two Method has been developed for the Manual Estimation of Total Red Blood Cell Count Using Hemocytometer / Neubauers chamber , MICRODILUTION METHOD FOR THE ESTIMATION OF TOTAL RBCs USING HEMOCYTOMETER, Materials Required for the Total Red Blood Cell (RBC) Count by Microdilution Method , Procedure of the Total Red Blood Cell (RBC) Count by Microdilution Method, MACRODILUTION METHOD FOR THE ESTIMATION OF TOTAL RBCs USING HEMOCYTOMETER, Materials Required for the Total Red Blood Cell (RBC) Count by Macrodilution Method, Procedure of the Total Red Blood Cell (RBC) Count by Macrodilution Method, Using Micropipette instead of RBC pipette for charging the Hemocytometer, CALCULATIONS FOR THE TOTAL RBC COUNT USING HEMOCYTOMETER, CALCULATIONS FOR TOTAL RED BLOOD CELL COUNT, HOW TO PERFORM TOTAL RED CELL COUNT IN LABORATORY, PRINCIPLE OF RED BLOOD CELL COUNT USING HEMOCYTOMETER, TOTAL RED BLOOD CELL COUNT BY HEMOCYTOMETER, TOTAL RED CELL COUNT USING NEUBAUER's CHAMBER, WHAT IS THE PURPOSE OF TOTAL RED BLOOD CELL COUNT, SPREAD PLATE CULTURE TECHNIQUE FOR THE ISOLATION OF MICROORGANISM / BACTERIA IN PURE CULTURE, STREAK PLATE CULTURE TECHNIQUE FOR THE ISOLATION OF MICROORGANISM / BACTERIA IN PURE CULTURE, Understanding The Postpartum Challenges And Finding Solutions, 7 Essential Ways to Improve Your Quality of Life Using Male External Condom Catheters. The number of RBCS per mm3 of the undiluted blood is then determined by this method. Your email address will not be published. the Decline in the no. The same pipette should be filled with RBC diluting fluid (preferably Hayem Fluid) until it reaches the mark 101. My greatest hobby is to teach and motivate other peoples to do whatever they wanna do in life. The most commonly used hemocytometer is the Improved Neubauer Chamber. The red blood cell count on the routine CBC is the concentration of red blood cells, expressed in millions/L of whole blood. Instead, take 4ml of the Diluting Fluid with the aid of a 5ml Graduated pipette and then discard 20 ul using either a micropipette (or RBC pipette). Focus the ruling with the 10x Objective Lens and then count the RBCs in five small squares of each square using the 40x Objective Lens. mm of central area is broken into 25 pieces so that the area is 25 squares = 1 sq. Place the RBC pipette in the container, and mix the solution again. Total RBC Count - This video explains the procedure of Red Blood Cell Count practical using Neubauer Chamber or Haemocytometer with calculations. [1] The hemocytometer was invented by Louis-Charles Malassez and consists of a thick glass microscope slide with a rectangular indentation that creates a precision volume chamber. This is a special type of glass chamber that is used for the cell counting, especially for Blood cells. The full grid on a hemocytometer contains nine squares, each of which is 1 mm 2 (Figure 3). I will appreciate your kind help. However, WBC(White blood cells originate in the bone marrow and circulate in the bloodstream and lymphatic system.) Counting Cells in a Hemocytometer. The Impact of Cerebral Telemedicine on Treatment Outcomes. of diluted blood.Cells in 1 mm cub. One may count the RBCs in 5 squares under low power and then under high power for comparing the results. To test your knowledge on this, you can take this hemocytometer quiz. This is especially true for cerebrospinal fluid. Haemocytometer "Neubauer"chamber is counting chamber with a cover slip. Before loading the RBC sample into the haemocytometer, discard 1-2 drops. The Neubauers Chamber has ruled the area of total 9 square mm and the depth is 0.1 mm as when the coverslip is placed on the surface of the counting chamber, the space between the bottom of the cover glass and the base of grooved area measures 0.1 mm in depth. Through a haemocytometer, RBCs are counted manually. and much more expensive than an average glass slide it can be used to count the number of red blood cells in a . With a micropipette draw approximately 20 ul of the diluted specimen. Required fields are marked *. of diluted blood are \"X\" x 50. So lets start with Microdilution method and then well move to Macrodilution method. Although a variety of automated cell counting instruments have been developed,Hemocytometerremains the most common method used for cell counting around the world. The first two drops of diluted blood from the pipette are discarded and then this mixture is run on the hemocytometer slide on both the chambers on both sides under a special coverslip.6. To make a hanging drop at your micropipettes tip, press the knob. The cells in four groups of 5 squares each i.e., a total of 80 squares.Red Blood Cell Count Calculation:Out of these 25 squares, we have counted the number of Red blood cells only from 5 boxes.Hence, total we have counted 16 x 5 = 80 squares.It is known that that 80 small squares have a volume of 1/50 mm cube. Other anticoagulants like sodium citrate, potassium oxalate etc., can also be used to prepare whole blood samples. Fill the same pipette with the RBC diluting fluid (preferably Hayems Fluid) up to the mark 101. Lets calculate total WBC count by using Neubauer counting chamber. It has a smooth surface and is even thicker than the chamber. We can conclude that the RBC or erythrocyte count method provides the concentration of RBCs per L of whole blood. The region which is count will differ for every kind of cell which is count and is calculate by means of the proportions of the ruled/lined region. Now, put the RBC pipette, mix the solution present in it again and then discard 1-2 drops from the pipette before charging the chamber. When counting cells that overlap an exterior line or ruling, count only those cells on the top or . From these 25 medium squares, only the big corner squares and the center squares inside the big center square are used to do RBCs counts. Then, wipe the RBC pipettes tip using blotting paper. Save my name, email, and website in this browser for the next time I comment. The central counting area of the hemocytometer contains 25 large squares and each large square has 16 smaller squares. Neubauers chamber is a thick glass plate with the size of a glass slide (30x70x4mm). Haemocytometer or Neubauers chamber slide is a manual method to count RBCs. A volume of 10 ml is sufficient to fill one counting chamber. The four corner squares are divided into 16 equal sized squares. In this exercise we will count the yeast present in the central large square. The central area is the 1 sq. 2. You should cover the Neubauer chambers central area or the ruled portion of Neubauers chamber with the glass cover to count the number of eukaryotic cell. Apparatus required:1) Neubauer Blood Counting Chamber: https://amzn.to/2S9VpR22) RBC Pipette: https://amzn.to/3xutNoD3) Red Blood Cell diluting fluid: (Hayem's fluid): https://amzn.to/3vFj75chttps://amzn.to/3zAE0ln4) Compound microscope: https://amzn.to/2U6Lj3S5) 90% ethyl alcohol: https://amzn.to/3xsJpZS6) Sterile cotton: https://amzn.to/3iOKdEa7) Sterile needle: https://amzn.to/2U6LuMA8) Reagent Bottles: https://amzn.to/3xvRBZj1.) Haemocytometer has a size of 30 X 70 X 4 mm. While red blood cell counts can be performed by manual techniques, such as a hemocytometer, these are time-consuming and inaccurate. Take out the Neubauers chamber / Hemocytometer from its case and clean it using a swab or gauze piece. Enter your email address and name below to be the first to know. The average size of Red Blood Cells (RBCs), is between 7.2 and 7.4 mm (microns). The ruled/lined zone is 3mm2 allocated into Nine (9) big squares, every one with a 1mm 2 area/zones. One colleague who used Nageotte hemocytometer wondered my decision, since he emphysized his support from Lutz publication at Transfusion Vol 33 No 5 Pages 409-412, 1993. The dilution is 1:20. The final pH of the solution at 25C varies between 5.8 and 6.0 depending on the composition and the companies that make it. It was invented by Dr Heinrich Schnitger. You can collect capillary blood by simply piercing the tip with a ring finger. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); 2022 LaboratoryInfo.com. Red blood cell count by neubauer chamberFor detailed information visit: https://www.ebiologylab.com/experiments/rbc-countPrinciple of Red Blood cell count Test:The total number of RBCs per mm3 of blood is determined by diluting known volume of blood 200 times with fluid, which prevents hemolysis and coagulation. 0.4mm3 of wbc squares contain wbcs=N RBC count and Hemoglobin estimation were done using hemocytometry using Neubauer's chamber with freshly prepared Hymes' diluting fluid and Sahli's Method respectively. The Area of five small squares equals 5/25, i.e. Shedding Light on SAD: Psychology of Seasonal Depression! Sampling bias is his concern. 1. You can also use a micropipette instead of RBC pipette for charging the Hemocytometer. What is the Difference Between Neisseria Gonorrhoeae and Neisseria Meningitidis, What is the Difference Between Ebola and Zika, Difference Between Classical, Alternative and Lectin Pathway, Difference between Biochemical Oxygen Demand (BOD) and Chemical Oxygen Demand (COD), Difference between Biomagnification and Bioaccumulation, Platelets (Thrombocytes) Definition, Structure, Function, Intrinsic Pathway of Apoptosis Definition, Process, Extrinsic Pathway of Apoptosis Definition, Mechanism, Functions, Regulation. Subsequently a small capacity of the diluted mixture of specimen/sample is counted, a common formula needed used to convert the cells counts into the no. If it is too dilute, the sample size will not be enough to make strong inferences about the concentration in the original mixture. Gently press the rubber tube of the RBC pipette, so that the next drop of fluid is in hanging position. By the help of micro-pipette, cautiously draw/pull up about 20ml of the cell dilution/mixture. I just purchased 3 Neubauer Chambers. <> On the surface of the middle platform, there are two sets of ruled area. Methodology Put the cover slip or glass slip on the top of grid area in the Chamber (use air tight technique) Dilute you sample: 1: 20 for WBC count 1:200 for RBC count and platelets Load your sample into the laoding area in the chamber Count the cells in the 4 large squares for WBC calculate the number of cells counted / L Required fields are marked *. The large central square (which can be seen in its entirely with the 10X objective), is divided into25 medium squareswith double or triple lines. However, a cell count can nowadays be carried out both manually and automatically [1, 3], and it is important to know where the limits and. After charging, wait for 3-5 min so that the cells settle down in the chamber . RBCs normal concentration in children, women and men range between 4.0-5.5 million/mcL, 4.2-5.4 million/mcL and 4.7-6.1 million/mcL, respectively. If you have any medical conditions that can cause high levels of red blood cells, tell your doctor about them. These squares have an area of 1 mm2 each.
The counting grid has a size of 3 mm X 3 mm. the increase in the no. This central segment is used for Red Blood Cell counting. A diagram below represents the pattern to count RBCs in all the five medium squares of a large central square. Other health or lifestyle factors can also lead to a high red blood cell (RBC) count. RBC Counting AreaThe large center square is used for RBC counts. An example of data being processed may be a unique identifier stored in a cookie. The Normal Saline solutions can also be used for this but it causes the slight creation of red blood cells and allows rouleaux formation which may cause the errors in results. Example: 50 sperm are counted in the five small squares. The Hayems fluid is isotonic to the Red blood cells and does not cause any damage to it. mN$ee2W7OY,Rs0] Your email address will not be published. Seven Ways to Adopt a Healthier Lifestyle, How to Instantly Fall Asleep After Drinking Coffee? Laboratories use two types of RBC Diluting Fluid today. The glass cover is placed on the top of the Neubauer chamber, covering the central area. However, in case you have to use it, be cautious that you should not intake the diluting fluid or Specimen. Which of the following can be used for counting? The Blog is basically devoted to the Paramedical personnels who risk their life to save the life of other peoples. Neubauer improved cell counting chamber. Suppose, N or number of RBCs in the five squares is 486, then the equation will be represented as: Total RBCs = 486 X 10,000 = 48, 60, 000 cells/L. CVs increased as the sampling area decreased. The central square is used for platelets and red cell count. 4. The one-counting chamber can be filled with 10 ul. We do, however, use them for counting RBC in fluids with low cell counts . Sperm/ml = 50 x 20/0.02 mm 3 x 1000 mm 3 /ml = 50,000,000 . If necessary, slowly expels the liquid from the chamber. Lets call it N number. Here is a way to determine a particle count using a Neubauer hemocytometer. Red blood cells (RBCs) are the round shape, biconcave discs, present in the blood that helps in the transport of gases throughout the body. What can nurses do to reduce the risk of medical malpractice? You can adjust the thresholds or sensitivity to prevent platelets, which are smaller than red blood cells, from being counted. Place the Neubauer chamber on the microscope stage. of RBC to more than 6.5 million/mm3) or Erythrocytopenia or Erythropenia (i.e. Blood Cross Matching Principle, Procedure, Result. These are counted in the four large corner squares labeled as WBC and if the number is Y. Count the cells that lie on the right and lower lines, but not on the opposite line. BLAUBRAND counting chambers are precision measuring instruments, used to determine the number of particles per volume unit of a suspension. After that, suck RBC diluting fluid or diluent upto a mark 101. As 10X is appropriate for WBC counting, count the total number of cells found in 4 large corner squares. Two methods have been developed to estimate total red blood cell count manually using Neubauers chamber or hemocytometer. Prick the ring finger after cleaning it with spirit swab. It is therefore necessary to dilute the blood specimen or blood sample using one of the RBC diluting liquids (hayems, formalin citrate diluting Fluid). CTRL + SPACE for auto-complete. White blood cell numbers (in the thousands/uL) cannot cause significant errors in red cell counts (in the millions/uL). The dilution factor used in the formula is determined by the blood dilution used in the cell count. 5 Types of Assisted Reproductive Technologies You Must Know Before Planning a Baby, The Science Behind Crackling Ears: Causes and Treatments. With the microscope, using a 4x objective, identify the nine main squares of the chamber delimited by three lines each as shown in the following image. Venous blood anticoagulated with EDTA, double oxalate or capillary blood Specimen requirement for RBC count 0.5 mark Draw blood in this mark of an RBC pipette 101 Draw the diluting fluid up to this mark Hayem's solution Diluting fluid used It has a round shape bulb which contains the Red bead to mix the blood specimen and the diluting fluid. Do not overcharge the chamber and there should be no air bubble in the Chamber. The blood is diluted with Hayem's fluid up to 101 mark by sucking the diluting fluid by the pipette kept in a watch glass. A hemocytometer is used to perform manual cell counts (RBC), nucleated cells, or platelets. The cells are then counted in areas that correspond to RBC count. In each square, you need to count the red blood cells located within the square. You should ensure that there is no air bubble inside the pipette bulb. After counting the cells under the microscope, we know the No.

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