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Feulgen Stain Kit 50ML For DNA Staining Reagents

  • G1048-50ML

Product Description

Principle

Feulgen stain is a staining technique used in histology to identify chromosomal material or DNA in cell specimens. It depends on acid hydrolysis of DNA in the 1-4 glycosidic bond, generating free aldehyde group. Then the aldehyde group combined with magenta forming purple-red complexes. DNA synthesis increase significantly during cell hyperplasia in pathological tissue (tumor, granulation). With Feulgen stain, the distribution (aggregate or disperse) and amount of DNA can be demonstrated.

Package content

Code

Name

Size

G1048-1

1 M hydrochloric acid

50ml

G1048-2

Magenta solution

50ml

G1048-3

Sodium metabisulfite solution

50ml

G1048-4

Fast-green solution

50ml

 

Preservation

Valid in 6 months at 4℃.   

 

Procedure  

1、Deparaffinize and hydrate sections to distilled water.

2、Preheat 1 M hydrochloric acid solution in 60℃ oven. Immerse slides in warm 1 M hydrochloric acid for 30 min, rinse in room-temperature1 M hydrochloric acid, wash with distilled water for 3 times.

3、immerse in periodic acid solution for 60-90 min, placed in dark condition. Do not wash in water. 

4、Rinse slides in 2 changes of Sodium metabisulfite solution, each 2 min. Wash in running tap water for 5 min. 

5、Rinse in fast-green solution for 10 s-2 min, dehydrate in 3 changes of absolute alcohol.

6、Clear in 2 changes of xylene, 5 min each. Mount with resin mounting medium.

7Microscopy determination and collect images for analysis.

 

Results

Nucleus DNA is purple-red while cytoplasm and other elements are light green.

Negative control

(1) Replace the hydrochloric acid in step 7, 8, and 9 with distilled water.

(2) After deparaffinize, incubate slides with DNase (1 mg/ml) at 37℃ for 2 h.

         Deal with either of the two ways above, slide is staining negative.

 

Notes 

1、Normal fix solution are alternative except Bouin solution which may result in over-hydrolysis of DNA. So Bouin solution is not suitable.

2、Too long or short time for hydrochloric acid hydrolysis both result in pale stain of nucleus. Generally, hydrolysis for 8 min is appropriate to most tissues. Adjusting the time according to hydrolysis result is recommended.

3、Manage the Fast-green counterstain in a proper range. Over-stain may cause deep blue or even overlap the positive results. 

G1048-50ML图片3Slice 10Slice 11Slice 12

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