Fluorescent microscope method for observing growing situation of plant pollen tube in pistil with high efficiency and high definition

A plant pollen, ingrowth technology, applied in fluorescence/phosphorescence, preparation of test samples, material analysis by optical means, etc. Tube and style structure, etc., to achieve the effect of improving microscopic imaging effect, improving transparency, and improving experimental efficiency

Active Publication Date: 2019-03-19
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some people also use NaClO for transparent treatment before softening, but no matter which way, after this kind of traditional method, it is impossible to avoid the strong autofluorescence of some strong styles with well-developed vascular tissue or self-tis

Method used

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  • Fluorescent microscope method for observing growing situation of plant pollen tube in pistil with high efficiency and high definition
  • Fluorescent microscope method for observing growing situation of plant pollen tube in pistil with high efficiency and high definition
  • Fluorescent microscope method for observing growing situation of plant pollen tube in pistil with high efficiency and high definition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Select the flowers of the tung tree 12 hours after pollination, peel off the petals, etc., leaving only the pistil part as the material to be tested, put the material to be tested into a 2.5mL centrifuge tube, add anhydrous sodium sulfite solution with a mass fraction of 10%, and use an amount of The material to be tested shall prevail, and then the centrifuge tube shall be placed in a water bath at 100°C for 2 hours and then taken out. The material to be tested will soften and become yellow and nearly translucent jelly-like. Aspirate the anhydrous sodium sulfite solution in the tube, wash carefully with distilled water for 3 times, then add 0.1% colorless aniline blue dyeing solution (preparation of 0.1% colorless aniline blue dyeing solution: weigh 0.1g aniline Blue is dissolved in 99.9g of K with a concentration of 0.033mol / L 3 PO 4 In the aqueous solution, prepare an aniline blue dyeing solution with a mass fraction of 0.1%, place it in the dark until decolorized, ...

Embodiment 2

[0031] Select the tung tree flower 12h after pollination, peel off the petals, etc., and leave only the pistil part as the material to be tested, and immerse in the FAA fixative solution (FAA fixative solution is composed of 50% ethanol aqueous solution with a volume fraction of 18:1:1, Glacial acetic acid and formaldehyde) were fixed for 4 hours, the fixed material to be tested was taken out, washed 3 times with distilled water, the material to be tested was placed in a 2.5mL centrifuge tube, and anhydrous sodium sulfite solution with a mass fraction of 10% was added. The material to be tested shall be submerged completely, and then the centrifuge tube shall be placed in a water bath at 90°C for 2 hours, and then taken out, the material to be tested will soften and become yellow and nearly translucent jelly-like. Aspirate the anhydrous sodium sulfite solution in the tube, carefully wash with distilled water 3 times, add 0.1% by mass fraction of colorless aniline blue dyeing so...

Embodiment 3

[0034]Select the wild peony flowers 12 hours after pollination, peel off the petals, etc., and leave only the pistil part as the material to be tested, and immerse in the FAA fixative solution (FAA fixative solution is composed of 50% ethanol aqueous solution with a volume fraction of 18:1:1, glacial acetic acid and formaldehyde) fixed in 8h, take out the fixed material to be tested, wash 5 times with distilled water, put the material to be tested in a 2.5mL centrifuge tube, add anhydrous sodium sulfite solution with a mass fraction of 10%, the dosage The material to be tested shall be submerged completely, and then the centrifuge tube shall be placed in a water bath at 100°C for 3 hours before being taken out. The material to be tested will soften and become yellow and nearly translucent jelly-like. Aspirate the anhydrous sodium sulfite solution in the tube, wash carefully with distilled water for 5 times, then add 0.1% by weight leuco aniline blue dyeing solution (preparation...

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Abstract

The invention discloses a fluorescent microscope method for observing the growing situation of a plant pollen tube in a pistil with high efficiency and high definition. The method can be used for greatly weakening the phenomenon of pistil tissue fluorescence and reducing disturbance of the pistil in pollen tube observation when the pistil is amplified by a relatively large factor, so that a high-definition image of growth in a pollen tube at high amplified factor can be easily obtained, and details are ideally represented. According to the fluorescence microscope method, the softening transparent time is greatly shortened to several hours, the dyeing time is shortened from several hours in the prior art to several minutes, an observation experiment can be completed in one day, and the experiment efficiency is greatly improved; and application of a high-concentration NaOH solution is prevented, the material size can be kept in the original state in the softening transparent process, thephenomenon of water-loss shrinkage cannot happen, and re-watering treatment is not required.

Description

Technical field: [0001] The invention belongs to the technical field of plant microscopy, and in particular relates to an efficient and high-definition fluorescence microscopy method for observing the growth of plant pollen tubes in pistils. Background technique: [0002] The growth of pollen tubes in the pistil is generally observed by fluorescence microscopy. The unique callose in the pollen tube is combined with the fluorescent pigment in the colorless aniline blue, which excites bright blue-green fluorescence under ultraviolet light, while other tissues in the pistil do not contain or contain little callose, so they do not emit light or weakly emit light. Through this technique, the growth of pollen tubes in the pistil can be effectively observed, and it is a necessary means for studying important physiological processes such as plant self-compatibility and fertilization process. [0003] The traditional pollen tube fluorescence microscopy technique simply softens and m...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N1/28G01N1/30
CPCG01N1/28G01N1/30G01N21/6428G01N2021/6439
Inventor 徐苑卿罗中莱张奠湘
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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