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PROCEDURE
placing a drop of blood from mixed sample on a clean glass slide. Spreader slide using another clean glass slide at 30-40 degree angle. Control thickness of the smear by changing the angle of spreader slide Allow the blood film to air-dry completely before staining. (Do not blow to dry. The moisture from your breath will cause RBC artifact.)
high HCT
large angle
tail
body
head
STAINING PROCEDURE
Thin smear are air dried. Flood the smear with stain. Stain for 1-5 min. Experience will indicate the optimum time. Add an equal amount of buffer solution and mix the stain by blowing an eddy in the fluid. Leave the mixture on the slide for 10-15 min. Wash off by running water directly to the centre of the slide to prevent a residue of precipitated stain. Stand slide on end, and let dry in air.
TOO
Too Alkaline Stain: 1. thick blood smear 2. 3. 4. prolonged staining insufficient washing alkaline pH of stain components Correction :
PRINCIPLE
White Blood Cells. 1. Check for even distribution and estimate the number present (also, look for any gross abnormalities present on the smear). 2. Perform the differential count. 3. Examine for morphologic abnormalities.
PRINCIPLE
Red Blood Cells, Examine for: 1. Size and shape. 2. Relative hemoglobin content. 3. Polychromatophilia. 4. Inclusions. 5. Rouleaux formation or agglutination
PRINCIPLE
Platelets. 1. Estimate number present. 2. Examine for morphologic abnormalities.
PROCEDURES
Observations Under 10 1. Check to see if there are good counting areas available free of ragged edges and cell clumps. 2. Check the WBC distribution over the smear. 3. Check that the slide is properly stained. 4. Check for the presence of large platelets, platelet clumps, and fibrin strands.
Observing direction:
Observe one field and record the number of WBC according to the different type then turn to another field in the snake-liked direction *avoid repeat or miss some cells
TIPS ON DIFF'S
Do not count cells that are disintegrating smudge cells eosinophil with no cytoplasmic membrane and with scattered granules Pyknotic cell (nucleus extremely condensed and degenerated, lobes condensed into small, round clumps with no filaments interconnecting). Basket cells
ABNORMAL DIFFERENTIALS
1. 200 Cell diff: a. WBC > 15.0 (>20.0 for babies under 1 month and labor unit) b. Three or more basophils seen. 2. If more than five immature WBC's are seen (or any blasts) let someone else diff slide and average results. 3. Correct WBC for NRBC's if you seen ten or more NRBCs/100 WBC. 4. Always indicate number of cells counted on diff. 5. If any cell type is extremely elevated (such as bands, monos, or eos > 20) indicate that you are aware of the abnormality by circling or checking on the card next to the results.
REPORTING RESULTS
Where possible use macrocytic and microcytic, rather than simply anisocytosis alone, when describing red cell morphology. Use specific cell morphology when possible, rather than simply reporting poikilocytosis. When red cells are normocytic, normochromic, report out as NORMAL. When abnormal morphology has been noted, DO NOT indicate normal on the report form. EXAMPLE: 7-10 microcytic RBC's/OIF is reported out as: 2+ microcytosis or Moderate microcytosis.
GRADING INCLUSIONS
STAB NEUTROPHIL
Diameter:12-16 Cytoplasm : pink Granules: primary secondary Nucleus: dark purple blue dense chromatin
BAND NEUTROPHIL
SEGMENTED NEUTROPHIL
Diameter: 12-16 Cytoplasm : pink Granules: primary secondary Nucleus: dark purple blue dense chromatin 2-5 lobes
SEGMENTED NEUTROPHIL
EOSINOPHIL
Diameter: 14-16 Cytoplasm : full of granules Granules: large refractile, orangered Nucleus: blue dense chromatin glass 2 lobes like a pair of
EOSINOPHIL
BASOPHIL
Diameter: 14-16 Cytoplasm : pink Granules: dark blue black obscure nucleus Nucleus: blue
BASOPHIL
LYMPHOCYTE
Diameter: small 7-9 large 12-16 Cytoplasm: medium blue Granules: small agranular granules large a few primary
LYMPHOCYTE
MONOCYTE
Diameter: 14-20 Cytoplasm : grey blue Granules: dust-like lilac color granules Nucleus: blue large irregularly shaped and folded
MONOCYTE
LEFT-SHIFT AND RIGHT-SHIFT OF NEUTROPHIL: Left-shift: non-segmented neutrophil > 5% Right-shift: hypersegmented neutrophil >3%
TOXIC GRANULATION
HYPERSEGMENTATION
Anisocytosis of neutrophil
vacuolizatio n
Degeneration of nucleus
Dohle body