Staining Flashcards
ROMANOWSKY STAINS
- founder
- produce multiple colors
- example of romanowsky stains:
- most commonly used
- for examination of malaria parasites
- for blood cell determination
- Dmitri Leonidovich
- POLYCHROMATIC STAINS
3.
- Wright
- Giemsa
- May Grunwald
WRIGHT’S STAIN: PREPARATION
1. wrights powder
2. acetone-free methanol
3. __ week at __ then RT/Ref temp
4. ____ / __ stain shortens the incubation
WRIGHT-GIEMSA STAIN: PREPARATION
1. wrights powder
2. giemsa powder
3. acetone-free methanol
4. age for ___ days
WRIGHT’S STAIN: PREPARATION
1. 1g
2. 500ml
3. 1 week at 37C
4. 1g Giemsa / 500 stain
WRIGHT-GIEMSA STAIN: PREPARATION
1. 9g
2. 1g
3. 2910ml
4. 30days
METHODS OF STAINING
1. makes use of a staining rack
2. more practical
- prepare ___ or ___ and fill them
- staining reagent (3)
- stains are combined
- stains are separated
3. automatic method follows the principle of ___ method, it’s just that it is performed by the machine.
- RACK METHOD
- DIP METHOD
- pulping jars or vessels
- FIXATIVE, EOSIN, METHYLENE BLUE
- GIEMSA STAIN
- WRIGHT STAIN - DIP METHOD
CELLS/ CELLULAR COMPONENT
1. takes up basic dye
- its nature is
- contains more acid like
2. takes up acid dye
- its nature is
- contains more basic like
3. react to both
- example
- BASOPHILIC
- ACID
- DNA - ACIDOPHILIC/ EOSINOPHILIC
- BASIC
- HGB - NEUTROPHILIC
- NEUTROPHIL
CRITERIA FOR A GOOD STAINS SMEAR
- macroscopic: blood smear should be ___ to _____
- microscopic
- Erythrocytes
- Platelets
- PMN
- Lymphocytes
- Monocytes
- Eosinophils
- Basophils
- pink to reddish-brown
- microscopic
- salmon pink
- purple-blue to lilac cytoplasm
- nuclei: purple blue; cytoplasm pinkish
- light blue cytoplasm
- faint blue gray cytoplasm
- red orange granules
- dark purple granules
CAUSES OF RED STAIN
1. excess ____ & excess ____
2. insufficient _____
3. very ____ smear
4. contaminants in _____
5. ___ before drying
6. _____ stain
CAUSES OF BLUE STAIN
1. ____ abnormality high is low HCT
2. heparinized blood sample
- wbc =
- ph of this anticoagulant is incompatible to stain so romanowsky stain gives ____ background
3. Avery high _____ with many _____
CAUSES OF RED STAIN
1. buffer; washing
2. staining time
3. thin
4. wash water
5. mounting the coverslip
6. old stain
CAUSES OF BLUE STAIN
1. Protein abnormality
2. heparinized blood sample
- gritty appearance
- dark blue or dark
3. LEUKOCYTE; blasts
Compositions of Romanowsky Stains:
- cationic/basic stain
- oxidation product (azure a)
- oxidation production (azure b)
- third oxidation product - anionic/acidic
- neutral component
- pH where ionization takes place
- decrease pH will produce
- increase pH will produce - fixative for 1min only as longer time causes thick smear
- Methylene Blue
- dimethyl thionine
- trimethyl thionine
- tetramethyl thionine - Eosin B/Y
- Thiazine-Eosinate Complex
- pH 6.4-6.8
- reddish color
- bluish color - methanol