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An optimized method for making paraffin sections of different floral tissues of Phalaenopsis

A production method and paraffin sectioning technology, which is applied in the preparation, sampling, and instrumentation of test samples, can solve the problems of less observation of flower stalks and petals, and achieve short production methods, save time, and maintain integrity Effect

Active Publication Date: 2020-10-27
ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are few reports on paraffin sections of Phalaenopsis, and most of them focus on embryogenic callus and leaf tissue, while there are few observations on flower parts such as pedicels and petals.

Method used

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  • An optimized method for making paraffin sections of different floral tissues of Phalaenopsis
  • An optimized method for making paraffin sections of different floral tissues of Phalaenopsis
  • An optimized method for making paraffin sections of different floral tissues of Phalaenopsis

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Experimental program
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Embodiment 1

[0046] The making of embodiment 1 Phalaenopsis petal tissue paraffin section

[0047] 1. Fixing: Wash the Phalaenopsis petals, take fresh tissues and fix them in FAA fixative solution containing 50% alcohol. Aspirate the fixative until no more air bubbles are produced to remove intercellular air from the material. After the fixation time is more than 24 hours, take the tissue out of the fixative solution and use a scalpel in the fume hood to smooth the tissue at the target site, thin the tissue to 2-3mm, and put the trimmed tissue into the dehydration box.

[0048] 2. Dehydration: Put the dehydration box into the hanging basket and dehydrate with gradient alcohol in the dehydrator. The alcohol gradient and dehydration time are as follows: 75% alcohol for 4 hours-85% alcohol for 2 hours-90% alcohol for 2 hours-95% Alcohol for 1 hour-absolute ethanol for 30 minutes-absolute ethanol for 30 minutes-alcohol benzene for 5 minutes-xylene for 5 minutes-xylene for 5 minutes.

[0049]...

Embodiment 2

[0055] The making of embodiment 2 Phalaenopsis non-lignified pedicel tissue paraffin section

[0056] 1. Fixing: Clean the non-lignified pedicel (young pedicel) of Phalaenopsis, take fresh tissue and fix it in standard fixative solution (FAA fixative solution) containing 70% alcohol. Aspirate the fixative until no more air bubbles are produced to remove intercellular air from the material. After the fixation time is more than 24 hours, take the tissue out of the fixative and use a scalpel in the fume hood to flatten the tissue at the target site, thin the tissue to 2-3mm, and put the trimmed tissue into the dehydration tray.

[0057] 2. Softening: Submerge the embedding frame with ethylenediamine softener, seal it, and put it in a blast drying oven at 55°C for softening. The softener replacement cycle is 7 days, and the softening degree is observed once a week, and the softening time is about 14 days.

[0058] 3. Dehydration: Put the dehydration box into the hanging basket a...

Embodiment 3

[0060] The making of embodiment 3 Phalaenopsis lignified pedicel tissue paraffin section

[0061] Referring to Example 2, wherein the softening time of step 2 is about 30 days.

[0062] Phalaenopsis lignified pedicel tissue sections stained blue-green ( image 3 ), which can be clearly distinguished from petal tissue and unlignified pedicel tissue.

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Abstract

The invention provides an optimized manufacturing method of paraffin sections of different flower tissues of phalaenopsis amabilis. The manufacturing method comprises the following steps: (1) fixing;(2) dehydrating; (3) waxing; (4) embedding; (5) slicing and unfolding; (6) dewaxing and rehydrating; (7) toluidine blue dyeing and microscopic examination, and carrying out proper differentiation anddrying according to tissue coloring depth; and (8) performing transparent sealing. Components and operation time of reagents in fixing, softening and dewatering links are optimized for different flower tissues of the phalaenopsis amabilis, the completeness of cells in slices is better kept, and unlignified soft tissues and lignified hard tissues can be clearly distinguished.

Description

technical field [0001] The invention belongs to the technical field of paraffin sections, and in particular relates to an optimized preparation method of paraffin sections of different floral tissues of Phalaenopsis. Background technique [0002] Paraffin section refers to the preparation of tissues by paraffin embedding, which is a commonly used way of preparation in histology. [0003] There are some deficiencies in the current conventional methods of making paraffin sections. One of them is that the cycle is long, it takes a long time to make a batch, and the procedures are complicated and there are many steps. The traditional paraffin section includes drawing, fixing, dehydrating, transparent, dipping in wax, embedding, slicing, spreading and sticking. , Dewaxing, staining, and mounting. In addition, in addition to the long cycle, each step is also related to each other at the same time. A mistake in a certain step will directly affect the observation results, such as ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30G01N1/36
CPCG01N1/30G01N1/36G01N2001/302G01N2001/305
Inventor 李佐肖文芳陈和明吕复兵
Owner ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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