Larva paraffin section forming method

A technology for paraffin sectioning and larvae, applied in the field of microscopic tissue sectioning, can solve the problems of difficulty in observing the complete spatial structure of worms, easy deformation or fragmentation of the digestive tract, limited tissue differentiation, etc., and achieves good transparent processing effect and tissue differentiation High-precision, short-time-consuming effect

Inactive Publication Date: 2014-04-30
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For this kind of larval body material with soft body, high liquid content and large cavity ratio, the current conventional slicing method is not only complicated to operate, but also takes a long time, and often uses toxic xylene and other chemical substances. The effect of final fixation, embedding, slicing, etc. is not ideal, and the final section has a very limited degree of tissue differentiation, especially the digestive tract, which accounts for a very large proportion of worms, is easily deformed or broken, and it is difficult to observe the complete spatial structure of worms

Method used

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  • Larva paraffin section forming method

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Embodiment 1

[0060] In the present embodiment, the paraffin section method of insect larvae comprises the following steps:

[0061] 1. Tissue fixation

[0062] Prepare a 4% paraformaldehyde solution in a chemical fume hood as a tissue fixative, and the fixative is prepared for immediate use. The preparation process of the tissue fixative is: in 100 ml of PBS buffer (PBS buffer: NaCl 137mmol / L, KCl 2.7 mmol / L, Na 2 HPO 4 10mmol / L, KH 2 PO 42mmol / L, pH value 5~7), add solid sodium hydroxide particles to make the pH value reach 11, then heat to 70°C in a microwave oven, add 4g of paraformaldehyde particles, shake quickly until dissolved, and use an ice water bath Cool the solution, adjust the pH value to 7 with concentrated sulfuric acid, finally add 30 μl Tween20, and cool the prepared tissue fixative solution to 4°C;

[0063] The fixed procedure is as follows: Put the tested worms (3rd instar larvae of cotton bollworm indoors, breeding room temperature 27°C, relative humidity 60%, lig...

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Abstract

The invention discloses a larva paraffin section forming method, and belongs to the technical field of microscopic tissue sections. The method comprises the following steps: fixing tissue, dehydrating the tissue, clearing the tissue, soaking with paraffin, embedding, slicing, unfolding, pasting, removing paraffin, and rehydrating the tissue. The operation is simple, and the consumed time is short; the tissue distinction degree of sections is high finally, and the complete space structure of a larva body can be observed; the method is suitable for larva body materials which are soft in body, high in liquid content and high in cavity proportion. When the method is adopted for processing a three-year cotton bollworm larva body, the body walls are complete when being observed at a bright field, and muscle tissues, nerve tissues and the like in body cavities are distributed, a digestive duct structure, which is in a large ratio of the body, is clear and complete, and a pair of pleopods are normal in space structure; the excitated spontaneous green fluorescence level of each organ tissue can be observed under blue light excitation.

Description

technical field [0001] The invention specifically relates to a method for paraffin sectioning insect larvae, and belongs to the technical field of microscopic tissue sectioning. Background technique [0002] Insect paraffin section technique is an important experimental technique for studying insect tissue morphology and histopathology, and generally includes steps such as dissection, fixation, dehydration and transparency, embedding, and sectioning. The exoskeleton of insects is relatively hard, so it is difficult to make complete paraffin sections. At present, insect paraffin sections are mostly used for local tissues or organs of insects (such as antennae, digestive tract, feet, etc.), so that the overall microstructure of insects cannot be observed. The flexible body wall of the insect larva wraps the open and flowing hemolymph and body fluid. The hemolymph and body fluid run through the tubular digestive tract from the head to the tail. The digestive tract is relatively...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N1/30
Inventor 李永丽周洲源春彦李红英刘圣明
Owner HENAN UNIV OF SCI & TECH
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