HematologyLab1Counting

Card Set Information

Author:
victimsofadown
ID:
265455
Filename:
HematologyLab1Counting
Updated:
2014-03-07 22:35:59
Tags:
Hematology
Folders:
Hematology
Description:
HematologyLab1Counting
Show Answers:

Home > Flashcards > Print Preview

The flashcards below were created by user victimsofadown on FreezingBlue Flashcards. What would you like to do?


  1. Describe the Unopette Diluting procedure in detail (nonspecific)
    • Separate shield from capillary pipette
    • Break diaphragm with pipette shield tip
    • Fill capillary tube to plastic intersection
    • Wipe outside of capillary tube to remove excess blood (NOT THE OPEN END)
    • slightly squeeze diluent reservoir
    • Place finger over plastic end of capillary tube
    • Introduce capillary tube into the diluent
    • Seat pipette firmly onto reservoir
    • Release pressure on diluent reservoir
    • Remove finger from plastic tip of capillary tube
    • Squeeze reservoir gently (rinse capillary tube) 2-3 times
    • Mix gently
    • Let stand for 10 minutes
    • REPEAT WITH A NEW UNOPPETTE
  2. Describe preparing the hemacytometer for charging with diluted sample (nonspecific)
    • Prepare a moist petri dish (wet filter paper and place 2 wooden sticks at bottom)
    • Clean hemacytometer surface and coverslip with alcohol and bleach
    • Rinse with distilled water
    • Dry w/ LENS PAPER only
    • Place dry coverslip on dry hemacytometer
    • Charge the hemacytometer
  3. Describe charging the hemacytometer in detail (nonspecific)
    • Invert unopipette several times
    • remove capillary tube from reservoir and reseat it in the reverse position (for use as dropper)
    • Wipe the exterior of the tube toward the tip to remove excess fluid
    • Squeeze reservoir gently and discard 3-4 drops
    • Place the capillary tube opening against the edge of the coverslip
    • Gently squeeze reservoir until diluted cells fill the chamber (evenly distributed)
    • Repeat on other side of chamber with other sample
    • Place charged hemacytometer in prepared petri dish and allow to sit for 10 minutes
  4. Describe an RBC determination (count) in detail
    • Dilute blood sample using RBC Unopette dilution system (.01mL blood / 1.99mL isotonic saline)
    • Dilution factor is 200
    • Repeat process using a new diluting system
    • Let the diluted samples stand for 10 minutes
    • Charge one side of the cleaned hemacytometer with one sample and repeat using the other diluted sample on the other side of the hemacytometer
    • Place the charged hemacytometer in the moist petri dish for ~10 minutes
    • Clean microscope w/ lens paper
    • Lower microscope stage and condenser to lowest positions
    • Place the charge hemacytometer on the microscope stage
    • Scan chamber under 10x objective until able to find grid and bring it into focus ("get bearings")
    • Using the 10x objective find the RBC counting area on the grid (CENTER PRIMARY SQUARE)
    • The four corner secondary squares and center secondary square will be used (TL, TR, BR, BL, M)
    • At 40x, count all cells in the 5 designated RBC squares (each contains 16 tertiary squares counted left to right, down, then right to left, down (repeated))
    • COUNT ONLY CELLS THAT TOUCH UPPER OR LEFT LINES
    • Keep an organized record of the # of RBCS in each secondary square, then total the cells
    • Repeat the same procedure on the other side of the hemacytometer
    • If the totals for both sides do not agree within 10% then dilutions/counts should be repeated
    • Determine the #RBCs for each side counted
    • (#counted x 10 x 200 / .2 / .2 /5)
    • Average the two results to determine the final result
    • Results are in RBCx106/μL or RBCx1012/L
  5. What is the generic equation for determining cell count?
  6. What are the normal values for an RBC determination?
    • Newborn: 4.4-5.8x106/μL
    • Adult male: 4.7-6.1x106/μL
    • Adult female: 4.2-5.4x106/μL
  7. Describe a WBC determination (count) in detail
    • Dilute blood sample using WBC Unopette dilution system (.02mL blood / 1.98mL ammonium oxalate and phosphate buffer)
    • Dilution factor is 100
    • Repeat process using a new diluting system
    • Let the diluted samples stand for 10 minutes
    • Charge one side of the cleaned hemacytometer with one sample and repeat using the other diluted sample on the other side of the hemacytometer
    • Place the charged hemacytometer in the moist petri dish for 10 minutes
    • Clean microscope w/ lens paper
    • Lower microscope stage and condenser to lowest positions
    • Place the charged hemacytometer on the microscope stage
    • Scan chamber under 10x objective until able to find grid and bring it into focus ("get bearings")
    • Using the 10x objective find the WBC counting area on the grid (FOUR CORNER PRIMARY SQUARES)
    • The 16 secondary squares in each primary square will be used (TL, TR, BR, BL)
    • At 40x, count all cells in the 4 designated WBC squares (each contains 16 secondary squares counted left to right, down, then right to left, down (repeated))
    • COUNT ONLY CELLS THAT TOUCH UPPER OR LEFT LINES
    • Keep an organized record of the # of WBCs in each primary square, then total the cells
    • Repeat the same procedure on the other side of the hemacytometer
    • If the totals for both sides do not agree within 10% then dilutions/counts should be repeated
    • Determine the #WBCs for each side counted
    • (#counted x 10 x 100 / 1 / 1 / 4)
    • Average the two results to determine the final result
    • Results are in WBCx103/μL or WBCx109/L
  8. What are the normal WBC determination values?
    • Newborn: 9.0-30.0x103/μL
    • Child/Adult: 4.8-10.8x103/μL
  9. Describe a platelet determination (count) in detail
    • **NOTE: ideally; count IMEDIATELY after WBC count is completed
    • Dilute blood sample using WBC Unopette dilution system (.02mL blood / 1.98mL ammonium oxalate and phosphate buffer)
    • Dilution factor is 100
    • Repeat process using a new diluting system
    • Let the diluted samples stand for 10 minutes
    • Charge one side of the cleaned hemacytometer with one sample and repeat using the other diluted sample on the other side of the hemacytometer
    • Place the charged hemacytometer in the moist petri dish for 10 minutes
    • Clean microscope w/ lens paper
    • Lower microscope stage and condenser to lowest positions
    • Place the charged hemacytometer on the microscope stage
    • Scan chamber under 10x objective until able to find grid and bring it into focus ("get bearings")
    • Using the 10x objective find the platelet counting area on the grid (ENTIRE CENTER PRIMARY SQUARE)
    • All 25 secondary squares in the primary square will be used
    • At 40x, count all cells in the 25 designated secondary platelet squares (each contains 16 tertiary squares counted left to right, down, then right to left, down (repeated))
    • COUNT ONLY CELLS THAT TOUCH UPPER OR LEFT LINES
    • Keep an organized record of the # of platelets in each secondary square, then total the cells
    • Repeat the same procedure on the other side of the hemacytometer
    • If the totals for both sides do not agree within 10% then dilutions/counts should be repeated
    • Determine the #platelets for each side counted
    • (#counted x 10 x 100 / 1 / 1 / 1)
    • Average the two results to determine the final result
    • Results are in plateletsx103/μL or plateletsx109/L
  10. What are the normal platelet determination  values?
    All: 130-400x103/μL
  11. What are the sources of error when performing a determination (any cell type)?
    • Blood sample (collected properly, free of clots)
    • Dilution (improper dilution)
    • Charging (too full/not full enough)
    • Counting (double counting, improper method)
    • Calculations (wrong dilution factors, etc)

What would you like to do?

Home > Flashcards > Print Preview