Staining and Smearing Hemo

1 hour or fix with methanol

EDTA

• make smear within 2-3 hours
• then fix with methanol if not stained within 1 hour

not generally used for smear

1/4 a drop

15-45^o

Decrease angle use faster motion.

• Increase angle.
• use slower motion.

Dirty slide

Pushes WBCs out to edges and distorts fragile cells.

• Unequal cell distribution.
• Platelet Clumping.

• Min length of 1inch.
• Marging on both sides.
• Even thickness.
• No Ridges, lines or holes.
• Feathered Edge.

• Lactophenol Cotton Blue
• Loeffler's Methalyene Blue
• Lugol's Iodine

• Stains background and not the cell.
• India Ink

• Stains Organism in Living State
• Organisms are not fixed(killed) before they are stained.

• Acridine Orange for Bacteria.
• Calofluor White for Fungus.
• Rhodamine-Auramine for TB

• 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

• Wright Stain
• Gram Stain
• AFB Stain

Distilled Water

• Methanol
• Cells are fixed to slide.

• Wright Stain
• Wright Giemsa
• May Greenwald
• Giemsa

Acidophilic Stain

Basophilic Stain

• Primary Stain-Eosin
• Counterstain- Methylene Blue
• Buffer-Distilled Water
• Fixative- Methanol

• –ALKALINE STAIN
• –PROLONGED STAIN TIME
• –INADEQUATE WASHING
• –BUFFER TOO ALKALINE
• –BLOOD SMEAR TOO THICK

Blood Started to Dry

Fat globules in blood

• Holes in Smear
• Possibly Hemoconia

Pressure on spreader slide

• Drop of Blood too small
• Angle of Spreader Slide
• Very Low patient hematocrit

• Drop of blood too large
• Angle of Spreader too big
• Very high Patient Hematocrit

Spreader slide was stopped near the end of the sample instead of pushed through.

• 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.

• Uneven motion or jerky
• Too much pressure on spreader slide.

2000 x cells per high power field

100

• 10
• Look for maturity, morphology , and inclusions

• 10 fields
• Size
• shape
• color
• inclusion
• maturity

20,000 times average of 10 oil immersion fields.

Increased central pallor

Decreased central pallor

No central pallor

Thin membrane with no center

normal color

• WNL- 05 total
• 1+ 1-2 in every oil field
• 2+ 1-3 in every field
• 3+ >3everyfield

• Sperocytes
• Target Cells
• Helmet Cells

Varience in shape of rbc

• Sperocytes
• Acanthocytes
• Sickle Cells
• Rouleaux

• 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)

• 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)

• Immature Reds
• Will stain with slightly bluish tint due to remaining RNA
• confirm with supravital stain