α-Naphthyl Acetate Esterase (α-NAE) Stain

Intended Use:

This kit is for staining bone marrow cell and bloodcell smear.

 

Principle:

Hydrochloric acid parafuchsin reacts with Sodium Nitrite to form diazonium salt (six-azo-pararosaniline).  α-Naphthyl Acetate is decomposed by esterase and produces α-naphthol, which shall combine with diazonium salt to form red brown precipitate in cytoplasm. This stain has no specificity against esterase; therefore, it is called "Non-specific Esterase Stain".

 

Specifications:

 

Contents	5Tests/Kit	20Tests/Kit	100Tests/Kit	Components
Fixative(solution A)	1vial×2.5ml	1vial×10ml	1vial×50ml	Sodium Citrate
Pararosaniline (solution B)	1vial×0.3ml	1vial×1.2ml	1vial×5.5ml	Parafuchsin
Sodium Nitrite (solution C)	1vial×0.3ml	1vial×1.2ml	1vial×5.5ml	Sodium Nitrite
PhosphateBuffer (solution D)	1vial×8ml	2vials×18ml	2vials×90ml	Phosphate
α-Naphthyl Acetate (solution E)	1vial×0.3ml	1vial×1.2ml	1vial×5.5ml	α-Naphthyl acetate
Methyl Green (solution F)	1vial×5ml	1vial×20ml	1vial×100ml	Methyl Green

 

Methods:

Working solution Preparation (for 1 test only):

Apparatus required: Disposable tube, micropipette, disposable tips, dropper;

Instruction: Add 50µl solution B and solution C in a test tube ; mix well and wait for 1 minute. Then, add 1.5ml solution D and 50µl solution E; mix well and wait for 2 minutes. (Add 1 drop of solution NaF for NaF inhibition test.)

	Solution B	Solution C	Solution D	Solution E	Solution NaF
Tube Α	50µl	50µl	1.5ml	50µl	--
Tube B (inhibition test)	50µl	50µl	1.5ml	50µl	1 drop

 

Expected Results:

Red or brown granules found in cytoplasm are positive.

①Spot pattern: It is mainly seen in mature T Lymphocytes. 1~4 brown or red-brown circular, mass, and big spot granules with clear contour  in cytoplasm are found in positive reaction. 

②Diffuse pattern: Red-brown and dusty  granules diffusely spread are found in positive reaction. Granules
may appear at a certain part of cell with obscure outline in cytoplasm. 

③Monocyte Pattern: Evenly stained red-brown granules diffusely spread in cytoplasm are found in
positive reaction.