Method for dissolving paraffin tissue sections, and application thereof

A tissue section and paraffin technique, applied in the field of dissolving and digesting paraffin-embedded tissue sections and dissolving paraffin tissue sections, can solve the problems of reduced DNA and RNA extraction, representative deviation, residual solid tissue at the bottom of the container, etc. Complete representation, the effect of shortening the time required for digestion and dissolution

Inactive Publication Date: 2017-10-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally, the proteinase K digestion incubation time for paraffin tissue nucleic acid extraction is 15 minutes to 3 hours, but for thicker and larger paraffin sections, even if the proteinase K incubation time is extended to 24 hours, it cannot be completely dissolved. solid tissue remains
Excessive extension of the incubation time will degrade some nucleic acid substances, and the existence of tissue residues will not only reduce the amount of DNA and RNA extracted, but also cause deviations in the representativeness of the DNA and RNA extracted from the dissolved and digested part of the tissue to the entire tissue, thereby Affect the measurement results

Method used

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  • Method for dissolving paraffin tissue sections, and application thereof

Examples

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Embodiment 1

[0020] see figure 1 , two breast cancer paraffin tissues cut from the same wax block were taken, labeled A and B, and the amount of tissue in the two tubes was basically the same. After the paraffin tissue sections in tube A and tube B were subjected to the same xylene dewaxing, alcohol cleaning and tissue drying, the tissue volume was about 5 mm×4 mm×6 mm (0.12 ml). Add 200 microliters of tissue lysate to each of the centrifuge tubes A and B containing the dry tissue to be lysed, and the formula of the lysate is a conventional SDS-containing solution. Add 15 microliters of proteinase K solution at a concentration of 20 mg / ml to centrifuge tube A, and add 25 microliters of the same proteinase K solution to centrifuge tube B to form a dissolution reaction system. Cap the centrifuge tube containing the lysis reaction system tightly and perform the first incubation on a metal bath thermostat at 56°C. After 60 minutes, the dissolution of the tissue was observed. It can be seen t...

Embodiment 2

[0024] see figure 1 , take a centrifuge tube containing a large number of paraffin sections of colorectal cancer tissue, after xylene dewaxing, alcohol cleaning and tissue drying, the tissue volume is about 8 mm × 5 mm × 10 mm (0.4 ml). Add 600 microliters of tissue lysate into the centrifuge tube containing the dried tissue to be dissolved, and the formula of the lyse is a conventional SDS-containing solution. Then add 25 microliters of proteinase K solution at a concentration of 20 mg / ml into the centrifuge tube to form a dissolution reaction system. Cap the centrifuge tube containing the lysis reaction system tightly and perform the first incubation on a metal bath thermostat at 56°C. After 60 minutes, observe the dissolution of the tissue, and it can be seen that the tissue in the centrifuge tube shrinks and settles at the bottom. Take out the centrifuge tube containing the dissolving reaction system and shake vigorously up and down to mix well, flick the bottom of the c...

Embodiment 3

[0027] see figure 1 , take a centrifuge tube containing gastric cancer paraffin section tissue, after xylene dewaxing, alcohol cleaning and tissue drying, the tissue volume is about 5 mm × 3 mm × 4 mm (0.06 ml). Add 200 microliters of tissue lysate to each centrifuge tube containing the dried tissue to be dissolved, and the formula of the lyse is a conventional SDS-containing solution. Then add 15 microliters of proteinase K solution at a concentration of 20 mg / ml into the centrifuge tube to form a dissolution reaction system. Cap the centrifuge tube containing the lysis reaction system tightly and perform the first incubation on a metal bath thermostat at 56°C. After 30 minutes, observe the dissolution of the tissue, and it can be seen that the tissue in the centrifuge tube shrinks. Take out the centrifuge tube containing the dissolved reaction system and shake it up and down vigorously to mix well, and flick the bottom of the centrifuge tube with your fingers to mix it eve...

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Abstract

The invention provides a method for dissolving paraffin tissue sections. The method comprises the following steps: heating an incubation dissolving system to a high temperature after routine primary protease K incubation is completed, keeping the temperature for 3-5 min, taking out obtained paraffin tissue sections, cooling the paraffin tissue sections to room temperature, adding protease K to a dissolving reaction system, and incubating the paraffin tissue sections for 10-30 min to completely dissolve and disappear the tissues. The method specially aims at a problem that thick or massive paraffin tissue sections cannot be thoroughly dissolved and digested within a short time. The method can completely digest and dissolve the massive thick paraffin tissue sections within 3 h to clarify a tissue solution without destroying nucleic acid substances and realize no obvious particles. The method remarkably shortens the time taken by the digestion and dissolving of massive thick paraffin tissue sections, no destroys to the nucleic acid substances, and reduction of nucleic acid substance degradation caused by long-time incubation, makes the extracted nucleic acid substances have a complete representative effect on the whole tissue sections, and can be applied to the dissolving of the paraffin tissue sections.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. The invention relates to the processing of paraffin samples of biological tissues, in particular to a method for dissolving paraffin tissue sections, which is a method for dissolving and digesting paraffin embedded tissue sections, and is especially suitable for use in extracting nucleic acid substances from paraffin samples of biological tissues. Background technique [0002] Tumor gene detection is currently a popular field of clinical testing. Through the detection of nucleic acid substances such as DNA, RNA, and miRNA in tumor tissue, the molecular characteristics of tumors can be obtained, so as to predict the clinical characteristics of tumor patients such as recurrence, survival, and treatment response. Paraffin-embedded tumor tissue sections are the most commonly used tumor gene detection samples. For example, the detection of EGFR mutation in paraffin-embedded tissue DNA of lung canc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
CPCG01N1/28
Inventor 黄彦钦彭佳萍葛维挺
Owner ZHEJIANG UNIV
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