Staining and Smearing Hemo

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Author:
moses1424
ID:
66624
Filename:
Staining and Smearing Hemo
Updated:
2011-02-16 17:16:23
Tags:
hemo
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Description:
hemo
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  1. CAPILLARY time limit.
    1 hour or fix with methanol
  2. Venous Collection Tube
    EDTA
  3. Venous collection time limit for stain
    • make smear within 2-3 hours
    • then fix with methanol if not stained within 1 hour
  4. ARTERIAL blood smear
    not generally used for smear
  5. How large of a drop of blood?
    1/4 a drop
  6. Angle of spreader slide?
    15-45o
  7. Drop is too large?
    Decrease angle use faster motion.
  8. If drop is too small?
    • Increase angle.
    • use slower motion.
  9. Open areas or uneven smear?
    Dirty slide
  10. Excessive Pressure while sliding?
    Pushes WBCs out to edges and distorts fragile cells.
  11. Excessive Delay causes?
    • Unequal cell distribution.
    • Platelet Clumping.
  12. Good Smear Criteria
    • Min length of 1inch.
    • Marging on both sides.
    • Even thickness.
    • No Ridges, lines or holes.
    • Feathered Edge.
  13. Simple Stains
    • Lactophenol Cotton Blue
    • Loeffler's Methalyene Blue
    • Lugol's Iodine
  14. Negative Stains
    • Stains background and not the cell.
    • India Ink
  15. Supravital Stains
    • Stains Organism in Living State
    • Organisms are not fixed(killed) before they are stained.
  16. Surpravital Stain Examples
    • Acridine Orange for Bacteria.
    • Calofluor White for Fungus.
    • Rhodamine-Auramine for TB
  17. Differential Stains
    • Uses 2 Stains and dyes that stain organisms different colors.
    • Allows for differentiation based on color and shape.
    • First dye is primary stain
    • second in the counterstain
  18. Differential Stain Examples
    • Wright Stain
    • Gram Stain
    • AFB Stain
  19. Buffer
    Distilled Water
  20. Fixative
    • Methanol
    • Cells are fixed to slide.
  21. Romanowsky Stain (differential Stain)
    • Wright Stain
    • Wright Giemsa
    • May Greenwald
    • Giemsa
  22. Eosin
    Acidophilic Stain
  23. Methylene Blue
    Basophilic Stain
  24. Wright Stain
    • Primary Stain-Eosin
    • Counterstain- Methylene Blue
    • Buffer-Distilled Water
    • Fixative- Methanol
  25. Wright Stain Source of Error
    • –ALKALINE STAIN
    • –PROLONGED STAIN TIME
    • –INADEQUATE WASHING
    • –BUFFER TOO ALKALINE
    • –BLOOD SMEAR TOO THICK
  26. Streak in Blood Smear
    Blood Started to Dry
  27. Hemoconia
    Fat globules in blood
  28. Dirty Slide
    • Holes in Smear
    • Possibly Hemoconia
  29. Uneven Distribution on Blood
    Pressure on spreader slide
  30. Smear too thin
    • Drop of Blood too small
    • Angle of Spreader Slide
    • Very Low patient hematocrit
  31. Smear too think
    • Drop of blood too large
    • Angle of Spreader too big
    • Very high Patient Hematocrit
  32. Smear ends in straight line instead of feathered edge.
    Spreader slide was stopped near the end of the sample instead of pushed through.
  33. Smear ends with numerous "tails"
    • Slide was pulled through drop of blood instead of just up to it.
    • Dirty edge on spreader slide.
    • Spreader slide wa turned so the edge of it was lifted off the sample edge.
  34. Smear contains ridges or waves
    • Uneven motion or jerky
    • Too much pressure on spreader slide.
  35. Estimated WBC count
    2000 x cells per high power field
  36. number of cells to count?
    100
  37. Oil Immersion WBC Number
    • 10
    • Look for maturity, morphology , and inclusions
  38. RBC oil immersion
    • 10 fields
    • Size
    • shape
    • color
    • inclusion
    • maturity
  39. Estimated Platelet Count
    20,000 times average of 10 oil immersion fields.
  40. Hypochromic
    Increased central pallor
  41. Hyperchromic
    Decreased central pallor
  42. Spherocytes
    No central pallor
  43. Anulocytes
    Thin membrane with no center
  44. Normochromic
    normal color
  45. Hypochromic limits
    • WNL- 05 total
    • 1+ 1-2 in every oil field
    • 2+ 1-3 in every field
    • 3+ >3everyfield
  46. Hyoerchromic Cells
    Examples
    • Sperocytes
    • Target Cells
    • Helmet Cells
  47. Poikilocytosis
    Varience in shape of rbc
  48. Shapes that are never normal.
    • Sperocytes
    • Acanthocytes
    • Sickle Cells
    • Rouleaux
  49. Never normal cell ranges.
    • 0 in normal
    • 1+ is 1-5 in 10 oil (1 in every other)
    • 2+ is 6-15 in 10 oil ( 1 in every)
    • 3+ is > 15 per 10 oil (1-2 in every field)
  50. All more normal shapes
    • 0-1 is wnl per oil
    • 1+ is 2-5 in 10 oil (1 in every other)
    • 2+ is 6-15 in 10 oil (1 in every field)
    • 3+ is >15 per 10 oil (1-2 in every field)
  51. Reticulocytes
    • Immature Reds
    • Will stain with slightly bluish tint due to remaining RNA
    • confirm with supravital stain

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