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    Hematology Laboratory: Manual RED Cell Counts

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    Ruel Antiola Mateo
    This document provides instructions for performing a manual red blood cell count using a hemacytometer. Blood is collected via fingerstick or venipuncture and loaded into a capillary pipette called a UNOPETTE. The blood is then diluted 1:200 in a diluting fluid like Hayem's or Gower's solution to fix and prevent agglutination of red blood cells. The diluted blood is mixed and loaded onto a hemacytometer slide for counting red blood cells under a microscope. Normal red blood cell count ranges in adults and newborns are provided. Potential sources of error in the technique include issues with equipment, technique, and inherent variability in cell distribution.

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    Hematology Laboratory: Manual RED Cell Counts

    Uploaded by

    Ruel Antiola Mateo
    34 views4 pages
    This document provides instructions for performing a manual red blood cell count using a hemacytometer. Blood is collected via fingerstick or venipuncture and loaded into a capillary pipette called a UNOPETTE. The blood is then diluted 1:200 in a diluting fluid like Hayem's or Gower's solution to fix and prevent agglutination of red blood cells. The diluted blood is mixed and loaded onto a hemacytometer slide for counting red blood cells under a microscope. Normal red blood cell count ranges in adults and newborns are provided. Potential sources of error in the technique include issues with equipment, technique, and inherent variability in cell distribution.

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    This document provides instructions for performing a manual red blood cell count using a hemacytometer. Blood is collected via fingerstick or venipuncture and loaded into a capillary pipette called a UNOPETTE. The blood is then diluted 1:200 in a diluting fluid like Hayem's or Gower's solution to fix and prevent agglutination of red blood cells. The diluted blood is mixed and loaded onto a hemacytometer slide for counting red blood cells under a microscope. Normal red blood cell count ranges in adults and newborns are provided. Potential sources of error in the technique include issues with equipment, technique, and inherent variability in cell distribution.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOC, PDF, TXT or read online from Scribd
    Download as doc, pdf, or txt
    34 views4 pages

    Hematology Laboratory: Manual RED Cell Counts

    Uploaded by

    Ruel Antiola Mateo
    This document provides instructions for performing a manual red blood cell count using a hemacytometer. Blood is collected via fingerstick or venipuncture and loaded into a capillary pipette called a UNOPETTE. The blood is then diluted 1:200 in a diluting fluid like Hayem's or Gower's solution to fix and prevent agglutination of red blood cells. The diluted blood is mixed and loaded onto a hemacytometer slide for counting red blood cells under a microscope. Normal red blood cell count ranges in adults and newborns are provided. Potential sources of error in the technique include issues with equipment, technique, and inherent variability in cell distribution.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOC, PDF, TXT or read online from Scribd
    Download as doc, pdf, or txt
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    Hematology Laboratory: Manual RED Cell Counts

    CLS 312
    Manual RED Cell Counts

    RBC Diluting Fluids

    1. Isotonic, so that cells are not hemolyzed or crenated.


    2. Must fix the cells to avoid autoagglutination.
    3. Hayem's Solution: (HgCl2, NaCl, Na2SO4, H2O)
    4. Gower's Solution: (Na2SO4, Acetic Acid, and H2O)
    UNOPETTE Procedure : RBC Counts

    1. Using the pipet shield of a 10 microliter capillary pipet, open the diaphragm of the dilution vial.

    2. Blood from fingerstick or venipuncture is allowed to fill the capillary of the UNOPETTE by capillary action.
    Filling of the capillary will stop when it is full.

    3. Wipe excess blood carefully from the outside of the


    pipet.

    4. Using one finger to cover the base of the pipet, insert the pipet tip first into the diluent reservoir while
    simultaneously squeezing the container.

    5. Release the reservoir and the end of the pipet simultaneously. this will draw the blood sample into the
    diluent. Blood is rinsed from the pipet by squeezing and releasing the diluent container several times.

    6. Cover the opening at the base of the pipet with a finger and invert the container several times to mix
    thoroughly.

    7. The base of the pipet can be reinserted into the vial for transportation and may be labelled on the outside.

    8. The pipet serves as the dispensing dropper for loading the hemacytometer.

    9. Discard the first three drops of mixture from the capillary prior to loading the hemacytometer.

    10. Place a clean hemacytometer cover glass over the chamber and allow the chamber to fill by capillary
    attraction between the cover glass and the unopette capillary. Count the cells.

    11. The dilution ratio for the RBC count is 1:200.

    12. Calculations:

    # Cells Counted X Dilution Factor X Depth Factor = # cells per cubic millimeter (mm3)
    # of Square Millimeters Counted
    Interpretation of Results

    Normals for a Red Count are as follows:

    Women 4.0-5.5 million/mm 3


    Men 4.5-6.0 million/mm 3
    Newborn 5.0-6.5 million/mm 3

    A decreased RBC count is seen in anemia, and an increased count is seen in polycythemia and dehydration.

    Sources of Error

    Errors in hemocytometry most frequently arise as a result of:

    1. apparatus
    2. personal technique
    3. inherent error

    (1) Errors caused by apparatus:

    1. chipped pipette tips


    2. obscure markings on pipettes
    3. non-optically plane cover glasses
    4. dirty glassware
    5. inaccurate rulings on chamber

    (2) Errors caused by personal technique:

    1. not thoroughly mixing blood


    2. inadequate shaking
    3. failure to discard first 4 drops
    4. not loading chamber properly (overfilling, trapped air bubbles)
    5. counting cells inaccurately (skipping cells, counting cells twice, counting on wrong borders)
    6. calculation error
    7. clerical error

    (3) Inherent errors in hemocytometry include:

    1. "field errors" - relates to the random distribution of cells on the counting chamber
    2. statistical error - occurs when total number of cells is too low to give statistical confidence in result
    (this error is reduced when larger numbers of cells are counted)

    47104042.doc
    Wednesday, December 01, 2010

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