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

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...

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.