Staining method of ciliates

A dyeing method and ciliate technology, applied in the preparation of test samples, etc., can solve the problems of loss of experimental samples, poor fixation effect, and experimental failure, and achieve the improvement of dyeing effect, better fixation effect, and shortening of experimental time. Effect

Active Publication Date: 2015-04-29
CHENGDU ACAD OF AGRI & FORESTRY SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the following disadvantages: 1. What fixation solution adopts is saturated mercuric chloride aqueous solution, because of solubility influence, fixation effect is not good; 3. In the process of fixing and washing, a large amount of experimental samples will be lost, and key structures such as cilia and cell openings are difficult to clearly display; 4. In the process of gradient alcohol dehydration, due to the external package There is collodion, and the dehydration time is long; 5. This method is highly dependent on the degree of technical mastery, and the operator needs to practice repeatedly to get a good dyeing effect: if the fixative is not washed thoroughly, the dyeing effect will be seriously affected. Improper timing of adding collodion, in the process of dehydration, it is easy to let all the successfully dyed samples come off with the collodion, resulting in the failure of the experiment

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1 (comparative example)

[0047] The traditional ciliate protein silver staining method, the steps are as follows:

[0048] (1) Front fixation: Use a straw to suck the worms to the corner of the glass slide to keep the residual moisture as small as possible, and then quickly wash the worms into the concave glass pit (pit) with saturated mercuric chloride (mercuric chloride) solution, Fix for 1-2 minutes (do not exceed 3 minutes).

[0049] (2) Rinse and patch: After fixation, rinse with warm distilled water for 3-5 times to wash off the mercury in the worm body. Suck out the water in the pit, leaving only a little water just enough to cover the worm body, add two drops of proteoglycerin water and mix well, and then suck the worm body onto a clean degreased glass slide. Suck off the excess protein glue, separate the worms from each other, add a drop of 0.5% collodion immediately after drying (when there is almost no moisture around the worm), shake it left ...

Embodiment 2

[0064] A method for dyeing ciliates, comprising the following:

[0065] Apply a layer of pure egg white evenly on one end of the slide and let it dry naturally.

[0066] Dip a little xylene saturated paraffin solution with a toothpick and spread evenly on the coverslip, and let it dry until the xylene is completely volatilized to form a uniform wax film on the coverslip.

[0067] Use a micropipette to suck the worm body to be sliced ​​to the corner of the egg white-coated area of ​​the glass slide to minimize the residual moisture, and then drop four drops of paste on the four corners of the wax-coated cover glass to cover the sample on the glass slide Above, lightly press the four corners of the coverslip with a toothpick to form a patch with worms fixed.

[0068] Pre-fixation: Add saturated alcoholic mercuric acid solution dropwise from one side of the coverslip as soon as possible, and fix for 1-3 minutes.

[0069] Take off the coverslip: put the patch into a staining jar...

Embodiment 3

[0083] The staining method of branchworm, including fixation in saturated alcohol mercuric solution and high-efficiency protein silver staining, specifically includes the following:

[0084] Preparation of protein glue glass slide: take pure egg white with a glass rod, evenly coat a layer of egg white protein on the right third of the glass slide, dry it in a desiccator for later use.

[0085] Use a secondary stretched micropipette to take a sample of Cladidae and place it in the embryo dish, rinse it with distilled water and place it in the middle of the protein-coated area on the protein glue glass slide.

[0086] Dip a little xylene saturated paraffin wax solution with a toothpick and apply it evenly on the cover glass. After the wax film is formed, suck the branch worm to the corner of the egg white-coated area of ​​the glass slide with a micropipette, so that the residual water is as little as possible. Quartz sand particles with a diameter slightly larger than the worm b...

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Abstract

The invention discloses a staining method of ciliates, belongs to the technical field of biology, and provides a staining method of ciliates to solve the problems that a traditional ciliate staining method is long in staining time and fussy in staining step, and cytostome and cilium cannot be clearly displayed. A saturated ethyl alcohol mercuric chloride solution is adopted as a fixing agent to carry out pre-fixation on ciliate bodies. In addition, before the pre-fixation operation, to-be-produced ciliate bodies are attached to a to-be-dyed glass slide; a plurality of steps such as dropwise adding protein glycerinum water and collodion are omitted. The staining method disclosed by the invention is convenient to operate; the sensitivity and the staining effect of silver staining of the ciliate protein are greatly improved; and rapid, efficient and accurate display of infraciliature in morphological classification and cytological studies of the ciliates is achieved.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a method for dyeing ciliates, in particular to a method for fixing and staining ciliates with protein silver. Background technique [0002] Ciliates use cilia as their motile organelles, and have cilia at least at some stage in their life cycle. If they do not have cilia, there is still a cilia system under the pellicle. Ciliates have two types of karyotypes, macronucleus and micronucleus, and have two modes of reproduction, namely, asexual reproduction by transverse bifisosis and sexual reproduction by conjugation. The lifestyle of ciliates is diverse and diverse. [0003] Ciliates are a class of organisms of great research significance. Ciliates are an important link between marine microfood rings and classic food chains, and play an important role in energy flow and species cycle; ciliates are harmful species in many aquaculture, and some species will cause red tides when ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
Inventor 齐桂兰吴永胜杨彪蒋泽银
Owner CHENGDU ACAD OF AGRI & FORESTRY SCI
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