a Acetic acetatease (ANAE) staining procedure.

asked: Who has a acetate sterase (ANAE) dyeing method of the specific steps, we can not find here, but also hope that you are not stingy to teach!
: Neutral non-specific lipase (α-NAE) dyeing
(Methodology) sterase dyeing methods are many, according to different effects such as pH substrates, the commonly used esterase staining method is divided into two categories, namely specific esterase staining and non-specific esterase dyeing.
the latter can be divided into 3 categories: (1) neutral nonse specific esterase dyeing method: α-acetate esterase, acetate esterase and acetate ass-D esterase. (2) Acidic nonse specific esterase staining method: α-acetate esterase and α-butylate esterase. (2) Alkaline non-specific esterase staining method: α-butylate esterase. Neutral non-specific esterase staining is an important method to distinguish between monocytes and granulocytic leukemia, which is available in general laboratories.
the principle is . α-acetate is hydrolyzed by esterase to produce phenol, which is coupled with heavy nitrogen salts to produce gray-black or brown-black sediments that are located in the plasma.

1, α - acetate acetone liquid: α - acetate 1g, acetone and distilled water mixture 100m1, dissolved and stored in a triangular bottle with abrasive plug, 4 degrees C storage, can be used for six months.
2, pH7. 4 Phosphate buffer: 0. 07MNa2HPO4 (i.e. Na2HPO4)
3, substrate: Phosphate buffer 82 ml, slow drop plus α-acetate propylene acetone liquid 1. 6ml (drip rate of about 40 drops per minute), while shaking hard. According to 5 minutes, add firm blue B80mg, shake vigorously for 10 minutes, immediately
filter
, filter two cups, 40 ml each.
4, sodium fluoride substrate liquid: called sodium fluoride (NaF) 61mg added to the above substrate liquid 1 cup, mixed well, marked.
the operation . (1) Fresh smears two, marked with NSE and NaF marks, drip plus 10% formaldehyde physiological saline, fixed for 5 minutes, with tap water gently rinsed, be careful of the blood membrane off, drying.
(2) two smears are placed in their respective dyeing cups at the same time, with a water bath of 37 degrees C for 1 hour.
(3) remove the water and wash, dry the mirror inspection. No re-dyeing.
the results are . In the cell pulp, if there is brown-black or gray-black diffuse particle precipitation is positive, the cell brain is yellow is negative.
the clinical significance of the . 1, positive: monocytes and
tissue
cells are strong positive, but can be inhibited by sodium fluoride;
2, weak positive and negative: granulocytes, lymphocytes are generally negative, if there is a weak positive can also be inhibited by sodium fluoride.
3, clinical identification: mainly used to identify acute monocyte leukemia and acute granulocytic leukemia, the former NSE positive, and inhibited by sodium fluoride (NaF), while the latter NSE negative, if positive is also not inhibited by sodium fluoride.
the note is . (1) Firm blue B and sodium fluoride can be sub-packed after
drying
save
(2) substrate liquid preparation process to shake, in order to promote the dissolution of the substrate. Warm up properly in winter preparation (with a water tank at 37 degrees C). The substation liquid is now useless, can not be saved, after filtration should be tested as soon as possible, so as not to precipitate a large number of sediments, affecting the dyeing effect.
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