Direct fluorescence labeling method for displaying cryptocaryon irritans brown microtubule cytoskeleton structure

A technology for stimulating Cryptonucleus and skeleton structure, applied in fluorescence/phosphorescence, determination/inspection of microorganisms, biochemical equipment and methods, etc. Simple, less time-consuming effects

Inactive Publication Date: 2012-07-25
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
View PDF1 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the steps of the above method are relatively complicated, the operation takes a long time, and the requirements for experimental conditions are relatively high.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Use a micropipette to draw 100 μL of 1% (V / V) polylysine aqueous solution, apply a circular area with a diameter of 1 cm on a clean slide, and air dry overnight for use;

[0032] (2) Under a Leica Wild M5 stereomicroscope, use a micropipette to draw 10 stimulated Cryptocaryon trophozoite cells in good condition, and transfer them into a container filled with 1 mL of PHEM buffer containing 0.5% (W / V) saponin. In a concave dish, infiltrate for 1 min;

[0033] (3) Absorb about 80% of the saponin solution in the previous step with a micropipette, then add 1 mL of PHEM solution to the concave dish with a micropipette and soak for 5 seconds;

[0034] (4) Absorb about 80% of the PHEM solution in the previous step, and then use a micropipette to transfer the infiltrated cells into another concave dish filled with 1 mL of 4% (W / V) paraformaldehyde solution, and fix for 20 seconds;

[0035] (5) Absorb about 80% of the paraformaldehyde solution in the previous step, then add ...

Embodiment 2

[0043] (1) Use a micropipette to draw 50 μL of 1% (V / V) polylysine aqueous solution, apply a circular area with a diameter of 1 cm on a clean slide, and air dry overnight for use;

[0044] (2) Under a Leica Wild M5 stereomicroscope, use a micropipette to draw 15 stimulated Cryptocaryon larval cells in good condition, and move them into the concave surface filled with 1 mL of PHEM buffer containing 0.5% (W / V) saponin In a dish, permeate for 1min;

[0045] (3) Absorb about 80% of the saponin solution in the previous step with a micropipette, then add 1 mL of PHEM solution to the concave dish with a micropipette and soak for 3 seconds;

[0046] (4) Absorb about 80% of the PHEM solution in the previous step, and then use a micropipette to transfer the infiltrated cells into another concave dish filled with 1 mL of 4% (W / V) paraformaldehyde solution, and fix for 10 seconds;

[0047] (5) Absorb about 80% of the paraformaldehyde solution in the previous step, then add 1 mL of PHEM sol...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a direct fluorescence labeling method for displaying a cryptocaryon irritans brown microtubule cytoskeleton structure, which comprises the following steps of: preparing a glass slide coated with polylysine; permeating cryptocaryon irritans brown cells through solution PHEM (Postherpetic Erythema Multiforme) containing saponin, and adding PHEM solution for immersion bath; fixing permeated cells through paraformaldehyde solution, and carrying out immersion bath through the PHEM solution, and transferring the fixed cells to the glass slide coated with the polylysine; dripping Triton X-100 water solution on the cells after removing extra fluid, carrying out immersion bath through the PHEM solution, and rinsing through 0.01mol / L of phosphate buffer; rinsing through 0.10mol / L of PBS (Phosphate Buffer Solution) after keeping from light, hatching through the PBS containing 1mu mol / L of fluorescence taxoid at room temperature, removing extra solution, and capping the glass slide; and finally, observing through a microscope and taking photos. The method is simple to operate, has high efficiency, is environmental-friendly and has no pollution.

Description

technical field [0001] The invention belongs to the field of methods for displaying and stimulating the microtubule cytoskeleton of Cryptocaryonia, in particular to a direct fluorescent labeling method for displaying and stimulating the microtubule skeleton structure of Cryptocaryon. Background technique [0002] Cryptocaryon stimuli, commonly known as "seawater melon worm", is a parasitic ciliate that parasitizes the body of bony fish in seawater. The body of the ciliate is distributed with cilia of uniform length, and these cilia are composed of microtubules. The life cycle of Cryptocaryon stimuli can be divided into three stages: trophozoite, cyst and larvae. The parasite was first discovered in 1937 by Sikama in an aquarium in Japan. In recent years, with the development of artificial breeding of seawater fish in our country, the increase of breeding density, and the mistakes of breeding management, stimulate the frequent occurrence of cryptocystosis in coastal provinc...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C12Q1/02
Inventor 尹飞施兆鸿彭士明孙鹏
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap