Tumor cell detecting method

A technology for tumor cells and detection methods, applied in biochemical equipment and methods, measuring devices, biological testing, etc., can solve problems such as fluctuations in the effect of cell nucleus staining, difficulty in grasping the time and degree of counterstaining, and affecting observation and judgment of results , achieve the effect of shortening the dyeing time, improving the dyeing effect and easy observation

Active Publication Date: 2017-01-04
HUNAN NEWLAND BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hematoxylin stains the nucleus, eosin stains the cytoplasm, any staining effect will affect the pathological diagnosis
In actual work, the staining effect of cell nuclei is prone to fluctuations, especially for tumor cells with heteromorphic nuclei, the time and degree of counterstaining are difficult to grasp, and it is easy to make the cytoplasm adhere to the color, and the background is not clean, which affects the observation and judgment of the results.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Detection of breast cancer cells

[0044] 1. Preparation of horseradish peroxidase (HRP) marker

[0045] (1) Dissolve 6mg of HRP in 1mL of 0.2mol / L pH4.5 sodium acetate buffer to make HRP solution;

[0046] (2) Add 0.2 mL of 0.1 mol / L sodium periodate solution to the HRP solution, and react in the dark for 30 minutes at room temperature;

[0047] (3) G25 gel column chromatography is used for the reaction solution of step (2), and 0.2mol / L carbonate buffer solution of pH9.5 is eluted, and the activated HRP is collected;

[0048] (4) Add 5 mg of alpha-fetoprotein (AFP) or ovalbumin (OVA) to the activated HRP solution, and react overnight at room temperature in the dark;

[0049] (5) Add 100 μl of freshly prepared 0.1mol / L sodium borohydride solution, and react in the dark for 30 minutes at room temperature;

[0050] (6) The reaction solution in step (5) is chromatographed on a G25 gel column again to collect protein elution peaks;

[0051](7) After concentr...

Embodiment 2

[0092] Example 2: Detection of lymphoma cells

[0093] 1. Preparation of HRP markers

[0094] (1) Dissolve 4mg of HRP in 1mL of 0.1mol / L pH4 sodium acetate buffer to make HRP solution;

[0095] (2) Add 0.1 mL of 0.15 mol / L sodium periodate solution to the HRP solution, and react in the dark for 10 min at room temperature;

[0096] (3) G25 gel column chromatography is used for the reaction solution of step (2), and 0.1mol / L carbonate buffer solution of pH 10 is eluted, and the activated HRP is collected;

[0097] (4) Add 1 mg of AFP or OVA to the activated HRP solution, and react overnight at room temperature in the dark;

[0098] (5) Add 50 μl of freshly prepared 0.05mol / L sodium borohydride solution, and react in the dark for 15 minutes at room temperature;

[0099](6) The reaction solution in step (5) is chromatographed on a G25 gel column again to collect protein elution peaks;

[0100] (7) After concentrating the solution in step (6), purify it with a G200 chromatograp...

Embodiment 3

[0115] Example 3: Detection of prostate cancer cells

[0116] 1. Preparation of HRP markers

[0117] (1) Dissolve 8 mg of HRP in 1 mL of 0.5 mol / L pH5 sodium acetate buffer to make HRP solution;

[0118] (2) Add 0.5 mL of 0.2 mol / L sodium periodate solution to the HRP solution, and react in the dark for 60 minutes at room temperature;

[0119] (3) G25 gel column chromatography is used for the reaction solution of step (2), and 0.5mol / L carbonate buffer solution of pH9 is eluted, and the activated HRP is collected;

[0120] (4) Add 10mg of AFP or OVA into the activated HRP solution, and react overnight at room temperature in the dark;

[0121] (5) Add 200 μl of freshly prepared 0.2mol / L sodium borohydride solution, and react in the dark for 45 minutes at room temperature;

[0122] (6) The reaction solution in step (5) is chromatographed on a G25 gel column again to collect protein elution peaks;

[0123] (7) After concentrating the solution in step (6), purify it with a G20...

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Abstract

The invention discloses a tumor cell detecting method, and belongs to the technical field of tumor cytology. The detecting method comprises the following steps of applying alpha fetoprotein with enzyme labeling or alpha fetoprotein heteroplasmon to combine with alpha fetoprotein acceptor heteroplasmon expressed by tumor cell; through the chromogenic reaction between substrate and enzyme, dyeing the tumor cell, and thereby distinguishing the tumor cell and the normal cell. The tumor cell detecting method can specially identify the tumor cell; the method overcomes the shortcomings of the traditional dyeing method, and is applicable to various tissue specimens or cell specimens; therefore, the method has very abroad application prospect.

Description

technical field [0001] The invention belongs to the field of tumor cytology, and in particular relates to a new method for detecting tumor cells. Background technique [0002] Clinically, the detection of alpha-fetoprotein (AFP) is an important indicator for the diagnosis of primary liver cancer, and it is also an important basis for evaluating the efficacy and prognosis. Studies have shown that embryonic and fetal cells can take up alpha-fetoprotein, and this uptake is selective and closely related to the degree of differentiation of cells. A large number of scientific research data show that the specific alpha-fetoprotein receptor mediates the uptake of alpha-fetoprotein, and this mechanism is regulated by cell differentiation. [0003] The concept of alpha-fetoprotein receptor was first proposed by Mriel, J in 1981, but earlier literature reports have indirectly indicated the existence of alpha-fetoprotein receptor-ligand system. Since then, similar receptors have been s...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/68G01N33/535C12Q1/02
CPCG01N33/5017G01N33/535G01N33/574G01N33/68
Inventor 宋光运汪媛媛
Owner HUNAN NEWLAND BIOTECH
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