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Method for slicing heavy leather blade paraffin

A paraffin section and leaf technology, which is used in sampling devices, preparation of samples for testing, etc., can solve the problems of difficult production process, broken materials, and prolonged soaking time of reagents.

Inactive Publication Date: 2011-09-07
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thick leathery blades present considerable difficulties in making due to the exceptional thickness of the blades
First of all, it is difficult for the reagent to enter the tissue, but the goal cannot be achieved by simply prolonging the soaking time of the reagent, because the soaking time is too long, which will bring adverse effects to the material. For example, soaking in xylene for too long will increase the brittleness of the material , the tissue is fragile
Secondly, the dehydration process is not easy, and if the dehydration is not complete, hollow wax flakes will be formed. Therefore, the paraffin slices of thick leathery leaves often have broken materials, resulting in the failure of the slices.

Method used

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  • Method for slicing heavy leather blade paraffin
  • Method for slicing heavy leather blade paraffin
  • Method for slicing heavy leather blade paraffin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Specific steps are as follows:

[0037] 1) Take fresh disease-free Cuban milk tree (Manilkara roxburghiana (Wight) Dubard) leaves (thickness 450~500μm, leathery), and cut 5~6mm on both sides of the main vein with a single-sided blade to cut the central veins And then cross-cut into small sections of 5-6mm.

[0038] 2) Fixation: The fixative adopts universal fixative (also known as FAA fixative, the formula is 70% ethanol 90ml, glacial acetic acid 5ml, formalin 5ml), fixed time 24h, and can be stored for a long time.

[0039] 3) Dehydration: dehydration using ethanol, starting with 60% ethanol, increasing by 5%, each level is 4h, specifically 60% ethanol 4h→65% ethanol 4h→70% ethanol 4h→75% ethanol 4h→80% ethanol Medium 4h→85% ethanol 4h→90% ethanol 4h→95% ethanol 4h→100% ethanol 4h (first time)→100% ethanol 4h (second time).

[0040] 4) Transparent: Take different ratios (volume ratio) of xylene and ethanol for transition, each level is 3 hours, the specific process is xylene:...

Embodiment 2

[0050] 1)~4) As described in Example 1.

[0051] 5) Use soft wax with a melting point of 52~54℃ for penetration wax, adopt a 1:1 (volume ratio) solution of xylene: paraffin wax to immerse the wax in a 37℃ thermostat for 72 hours, then change to pure paraffin wax in a 63℃ thermostat During the immersion of wax, it is necessary to change the pure paraffin one or two times, and then embed.

[0052] 6) As described in Example 1.

[0053] 7) Extend the film: first transition in 25% ethanol, and then extend the film in warm water at 55°C.

[0054] 8) As described in Example 1.

[0055] 9) Baked slices: Baked slices in a 35°C incubator for 36 hours.

[0056] 10) Dewaxing and rehydration: The specific procedure is 100% ethanol: xylene: 1:2 (volume ratio) solution for 1 min→100% ethanol: xylene: 1:1 (volume ratio) solution for 1 min→100% ethanol: Xylene is 2:1 (volume ratio) in a solution for 1 min→100% ethanol for 1 min (first time) → 100% ethanol for 1 min (second time) → 95% ethanol for 1 mi...

Embodiment 3

[0060] 1)~6) As described in Example 1.

[0061] 7) Extend the film: first transition in 35% ethanol, and then extend the film in warm water at a water temperature of 48°C.

[0062] 8) As described in Example 1.

[0063] 9) Baked slices: Baked slices in a 35°C incubator for 36 hours.

[0064] 10) Dewaxing and rehydration: The specific procedure is 100% ethanol: xylene for 1:2 (volume ratio) solution for 2 min → 100% ethanol: xylene for 1:1 (volume ratio) solution for 2 min → 100% ethanol: Xylene is 2:1 (volume ratio) in the solution for 2min→100% ethanol 2min (first time)→100% ethanol 2min (second time)→95% ethanol 2min→85% ethanol 2min→70% ethanol 2min→50 % Ethanol for 2min.

[0065] 11) Staining: the specific procedure is 1% safranin solution for 3h→95% ethanol for 2min→100% ethanol for 2min (first time)→100% ethanol for 2min (second time)→100% ethanol:xylene is 2:1 2min in the solution→100% ethanol:xylene for 2min in a 1:1 solution→100% ethanol:xylene for 2min in a 1:2 solution→xy...

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Abstract

The invention discloses a method for slicing heavy leather blade paraffin and relates to a method for slicing plant paraffin. Operating procedures include fixing, dehydrating, transparentizing, paraffin infiltration, embedding, slicing, dewaxing, dyeing and mounting. In the splicing process of the heavy leather blade paraffin, a fixing agent, a dehydrating procedure, a transparentizing procedure, a paraffin infiltration method, a dyeing method and mounting are improved, i.e., selection of the fixing agent, determination of the dehydrating procedure, determination of the transparentizing procedure, determination of the dewaxing procedure, the dyeing method and the mounting method are improved. The method is also suitable for other blades with different thicknesses and properties. By adopting the method, slice manufacturing of blade paraffin with different thicknesses and properties can be realized practically, and a uniform and practical manufacturing method for theoretical manufacturing of paraffin is popularized.

Description

Technical field [0001] The invention relates to a paraffin sectioning method for plants, in particular to a paraffin sectioning method for thick leathery leaves. Background technique [0002] Paraffin sections are widely used in histological routine preparation techniques, mainly used to observe the morphological structure of cell tissues, and can be stored for a long time. There are many procedures and procedures for paraffin section production, and the cycle is long. Each link will affect the quality of the section. For example, problems such as section bending and section cracks are prone to occur during the sectioning process (Wu Minhua, Ke Zhanhua. Common problems and solutions in the sectioning process in paraffin sectioning Method [J] Journal of Hainan Medical College, 2006, 12(3): 255). Although these problems can be overcome, if the material is broken, it cannot be remedied and the section must be remade. The cause of material fragmentation can be traced back to many st...

Claims

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Application Information

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IPC IPC(8): G01N1/06G01N1/28G01N1/30
Inventor 刘育梅卢昌义
Owner XIAMEN UNIV
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