An Improved Grocott Hexamine Silver Staining Method and Its Application

A dyeing method, the technology of hexaamine silver, is applied in the field of dyeing entomopathogenic fungi, which can solve the problems of peeling off and achieve excellent dyeing effect

CN104677713BActive Publication Date: 2017-07-04QINGDAO AGRI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2017-07-04

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Abstract

The invention discloses an improved Grocott silver hexosamine staining method and applications thereof. The staining method comprises the steps of preparing paraffin-embedded sections, staining, carrying out dehydrated hyalinizing, and carrying out section sealing, wherein in the step of preparing paraffin-embedded sections, a sliced paraffin section is adhered to a glass slide subjected to APES treatment. The method overcomes the defect that section shedding easily occurs in the process of staining, especially for frankliniella occidentalis with a small somatotype, the non-section-shedding rate is over 70%. In addition, the internal tissues of polypides in sections obtained according to the staining method disclosed by the invention are red, blastospores in polypides are black, therefore, the polypides are clearly visible, and the staining effect is excellent.
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Description

technical field

[0001] The invention relates to a staining method for entomopathogenic fungi, in particular to an improved Grocott hexamine silver staining method, which is particularly suitable for application in paraffin sections of Thrips occidentalis-Beauveria bassiana. Background technique

[0002] Western flower thrips Frankliniella occidentalis (Pergande) is a worldwide agricultural pest belonging to the order Thysanoptera, Thripidae, genus Flower Thrips. Its host range is relatively wide, and there are currently more than 500 species of plants in 60 families. Western flower thrips was first discovered in Beijing in 2003, and now it has become an important pest in vegetable production in open fields and greenhouses in my country. Western flower thrips are highly resistant to chemical insecticides, so it is very necessary to take biological control of western flower thrips. In particular, Beauveria bassiana infects insects through the body wall, which is extremely su...

Examples

Embodiment 1

[0051] The specific dyeing methods are:

[0052] (1) The production steps of paraffin tissue sections mainly include procedures such as material collection, fixation, washing and dehydration, transparency, wax immersion, embedding, sectioning and sticking, as well as dewaxing and hydration, among which:

[0053](a) The material taken is the adult thrips western flower 72h after inoculating Beauveria bassiana;

[0054] (b) The fixative adopts 10% neutral formaldehyde fixative;

[0055] (c) The slices were sliced ​​with a Leica microtome, and the thickness of the cut paraffin slices was 8 μm;

[0056] (d) The glass slide is treated with APES (3-aminopropyl-3-methoxysilane) before sticking the film, and the specific treatment method is as follows: wash the glass slide with a lotion (the preparation of the lotion is as follows: 20g dichrome Dissolve potassium acid potassium in 110mL of distilled water, then add 90mL of concentrated sulfuric acid drop by drop, and the speed of ad...