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    Cell Counting Areas in Neubauer Chamber

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    The document discusses manual cell counting using a Neubauer hemocytometer. The Neubauer chamber is a thick glass slide with ruled areas used to count cells under a microscope. It has squares divided into smaller squares to calculate cell concentrations in a sample. Different areas of the chamber are used to count white blood cells, red blood cells, and platelets. Cells are counted and the numbers are converted to a concentration value per microliter using the chamber's dimensions and sample dilution factor.

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    Cell Counting Areas in Neubauer Chamber

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    162 views4 pages
    The document discusses manual cell counting using a Neubauer hemocytometer. The Neubauer chamber is a thick glass slide with ruled areas used to count cells under a microscope. It has squares divided into smaller squares to calculate cell concentrations in a sample. Different areas of the chamber are used to count white blood cells, red blood cells, and platelets. Cells are counted and the numbers are converted to a concentration value per microliter using the chamber's dimensions and sample dilution factor.

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    readings and calculations

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    Neubar Chamber

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    The document discusses manual cell counting using a Neubauer hemocytometer. The Neubauer chamber is a thick glass slide with ruled areas used to count cells under a microscope. It has squares divided into smaller squares to calculate cell concentrations in a sample. Different areas of the chamber are used to count white blood cells, red blood cells, and platelets. Cells are counted and the numbers are converted to a concentration value per microliter using the chamber's dimensions and sample dilution factor.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    162 views4 pages

    Cell Counting Areas in Neubauer Chamber

    Uploaded by

    nice person
    The document discusses manual cell counting using a Neubauer hemocytometer. The Neubauer chamber is a thick glass slide with ruled areas used to count cells under a microscope. It has squares divided into smaller squares to calculate cell concentrations in a sample. Different areas of the chamber are used to count white blood cells, red blood cells, and platelets. Cells are counted and the numbers are converted to a concentration value per microliter using the chamber's dimensions and sample dilution factor.

    Copyright:

    © All Rights Reserved
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    of 4

    MANUAL CELL COUNTING WITH NEUBAUER CHAMBER

    Although a variety of automated cell counting instruments have been developed, Hemocytometer
    remains the most common method used for cell counting around the world. The most frequently
    used haemocytometer is the Neubauer (or „Improved Neubauer‟) chamber. Other
    haemocytometers include the Burker, Thoma and Fuchs-Rosenthal. Using these, the particles
    (e.g., leucocytes, erythrocytes, thrombocytes, bacteria, fungus spores, pollen) are visually
    counted under a microscope.

    GENERAL FEATURES OF THE NEUBAUER’S CHAMBER

    Neubauer‟s chamber is a thick glass plate with the size of a glass slide (30x70x4mm). The
    counting region consists of two square shaped ruled areas. There are depressions or the moats on
    either side or in between the areas on which the squares are marked thus giving an “H” shape.

    The ruled area is 3mm2 divided into 9 large squares each with a 1 mm2 area. The large central
    square (which can be seen in its entirely with the 10X objective), is divided into 25 medium
    squares with double or triple lines. Each of these 25 squares are is again divided into 16 small
    squares with single lines, so that each of the smallest squares has an area of 1/400 mm2.

    The glass cover is a squared glass of width 22 mm. The glass cover is placed on the top of the
    Neubauer chamber, covering the central area. The ruled area is 0.1 mm lower than the rest of the
    chamber. 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.

    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.

    WBC Counting Area


    The four large sqaures placed at the corners are used for white blood cell count. Since their
    concentration is lower than red blood cells a larger area is required to perform the cell count.

    RBC Counting Area


    The large center square is used for RBC counts. As already stated, this area is subdivided into 25
    medium sqaures, which in turn are each divided into 16 squares. Of the 25 medium sqaures, only
    the four corner squares and the center square within the large center square are used to perform
    RBC counts.

    Platelet Counting Area


    The large center square is used to count platelets. Platelets in all 25 squares within the large
    center square are counted.
    MICROSCOPE FOCUSING AND CELL COUNTING

    Place the Neubauer chamber on the microscope stage. Using the 10X objective, focus both
    onto the grid pattern and the cell particles.
    As 10X is appropriate for WBC counting, count the total number of cells found in 4 large
    corner squares.
    To count the RBCs and Platelets, the microscope must be switched to 40X objective. Count
    the cells in the respective areas as stated early.
    Write down the amount of cells counted

    If cells are touching the 4 perimeter sides of a corner square, only count cells on 2 sides, either
    the 2 outer sides or 2 inner sides.

    CALCULATING THE CELL COUNTS

    The total number of cells per microliter of sample can be calculated from the number of cell
    counted and area counted. This is because the ruled areas of the chamber contain an exact
    volume of diluted sample. Since only a small volume of diluted sample is counted, a general
    formula must be used to convert the count into the number of cells/microliter.

    The dilution factor used in the formula is determined by the blood dilution used in the cell count.
    The depth used in the formula is always 0.1. The area counted will vary for each type of cell
    count and is calculated using the dimensions of the ruled area.

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