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Manufacturing method of daphnia microslide specimen

A technology for microscopic slides and production methods, which is applied in the preservation, application, and animal husbandry of human or animal bodies. It can solve the problems of difficult to distinguish fine structures, long drying time of glycerin gel, and unclear observation, etc., and achieve the internal structure. Clear, shortened drying time, improved transparency

Inactive Publication Date: 2010-08-04
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for Daphnia large daphnia and Daphnia jianshui as whole specimens, it is not necessary to dehydrate, and the slides can be directly sealed with glycerin glue, which is a simple operation process, but there are some problems. Glycerin glue takes a long time to dry and is easy to slide, causing specimens shift
At the same time, for water fleas, the body is transparent, and the internal structure of the body can be directly observed through a microscope, but in general, the observation is not clear, and xylene is needed to make it transparent and then seal the film
Canada gum or neutral resin is often used for sealing slides. Although their technical parameters such as light transmittance, UV blocking, and refraction meet the requirements for sealing biological microscopic slide specimens, the intestines, ovaries, and oculomotor muscles in the body Difficult to distinguish such fine structures

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Fixation: live daphnia were anesthetized and fixed in 70% ethanol aqueous solution.

[0024] Single dyeing: Dyeing with borax magenta for 1 day.

[0025] Color separation: Hydrochloric acid-alcohol color separation, that is, add 1-2 drops of hydrochloric acid to 70% ethanol until no dye falls.

[0026] Ethanol dehydration step by step: 85% ethanol→95% ethanol→absolute ethanol dehydration (absolute ethanol 3 times) (15 minutes for each gradient).

[0027] Replacement: The ethanol in the dehydrated water flea body was replaced by 35% acetone → 50% acetone → 75% acetone → 85% acetone → 95% acetone → anhydrous acetone (anhydrous acetone 3 times) (15 minutes for each gradient).

[0028] Sealing: suck out the daphnia in acetone with a straw and put it directly on a clean glass slide. After a little evaporation, add 1 or 2 drops of pre-polymerized methacrylate, and carefully cover the cover glass to avoid air bubbles. After sealing the slides, place them in an oven at 60°C f...

Embodiment 2

[0036] Fixation: Live daphnia were placed in Bouin's fixative for 1 hour.

[0037] Dyeing: First, chrome alum cyanine dyeing, dyeing for 3 hours.

[0038] Color separation: hydrochloric acid - alcohol color separation.

[0039] Gradual ethanol dehydration: 35% ethanol → 45% ethanol → 55% ethanol → 75% ethanol → 85% ethanol → 95% ethanol → absolute ethanol → absolute ethanol → absolute ethanol dehydration, each gradient is 20 minutes.

[0040] Counterstaining: When dehydrating to 95% ethanol, prepare saturated orange G ethanol solution and dip for 1 hour.

[0041] Replacement: The ethanol in the dehydrated water flea body is replaced by 35% acetone → 50% acetone → 75% acetone → 85% acetone → 95% acetone → anhydrous acetone → anhydrous acetone → anhydrous acetone, each gradient is 15 minutes.

[0042] Sealing: suck out the daphnia in acetone with a straw and put it directly on a clean glass slide. After a little evaporation, add 1 or 2 drops of pre-polymerized methacrylate, an...

Embodiment 3

[0050] Live daphnia were placed in 4% neutral formaldehyde solution for fixation, and the rest was the same as in Example 2 to obtain a microscopic slide specimen of daphnia.

[0051] The water flea microslide specimens prepared in Examples 1, 2 and 3 were observed under an optical microscope respectively. The cell boundaries of the water flea were obvious, and structures such as the intestinal tract and ovary were observed more clearly, and the transparency was good.

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Abstract

The invention provides a manufacturing method of a daphnia microslide specimen, which comprises the following steps: firstly fixing the daphnia with a fixative, simple-staining or double-staining, color separating, and stepwise dehydrating with ethanol; then stepwise displacing the ethanol in the dehydrated daphnia with acetone; and finally sucking out the daphnia which is subject to the displacement with acetone, dripping on a glass slide, and dripping prepolymerized polymethacrylate to seal the glass slide to obtain a daphnia microslide specimen. The daphnia microslide specimen manufacturedby the method enables the cell borderlines of the daphnia to be obvious, the intestinal tract, ovary and other structures to be observed more clearly, the transparency is increased, and the drying time is shortened.

Description

technical field [0001] The invention relates to the preparation of biological specimens, in particular to a preparation method of a permanent water flea microslide specimen. Background technique [0002] The overall slide preparation methods currently used for microscopic slide specimens are: 1. Temporary and semi-permanent slide preparation methods, including glycerin method, glycerin jelly method and lactic acid-phenol method; 2. Permanent overall slide preparation methods, including Venetian turpentine method, tert-butanol gum method and dioxane gum method, etc.; 3. Amber specimen preparation method for larger insects, etc. However, for daphnia materials such as daphnia daphnia and daphnia jianshui as whole specimens, it is not necessary to dehydrate, and the slides can be directly sealed with glycerin glue, which is a simple operation process, but there are some problems. Glycerin glue takes a long time to dry and is easy to slide, causing the shift. At the same time, ...

Claims

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Application Information

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IPC IPC(8): A01N1/00
Inventor 王茜王兰张小民武玉珑姜美玲
Owner SHANXI UNIV
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