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A cell pretreatment method for electrochemical detection

An electrochemical and cellular technology, applied in the direction of electrochemical variables of materials, etc., can solve the problem that the release of electroactive substances is not complete enough, and cannot truly reflect changes in cell activity, so as to facilitate the detection of cell activity, improve sensitivity, and enhance electricity. Effects of chemical signaling

Active Publication Date: 2018-03-09
JIAMUSI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The present invention aims to solve the problem that the existing cell pretreatment technology does not fully release electroactive substances, so that it cannot truly reflect the change of cell activity, and provides a cell pretreatment method for electrochemical detection

Method used

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  • A cell pretreatment method for electrochemical detection
  • A cell pretreatment method for electrochemical detection
  • A cell pretreatment method for electrochemical detection

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specific Embodiment approach 1

[0011] Specific embodiment 1: In this embodiment, a cell pretreatment method for electrochemical detection is a cell in situ complex lysis method. 2 Cultivate in an incubator with a volume concentration of 5% for 24 hours, then remove the medium, add hypotonic lysate, and adjust the concentration of cells to be detected to 5×10 4 ~5×10 8 cells mL -1 , and then under the condition of 30-70°C water bath, crack for 30-60 minutes, which is complete.

[0012] This specific embodiment combines the hypotonic lysis method on the basis of the cell heating lysis method, that is, replaces the hypertonic PBS solution with a hypotonic solution such as double distilled water, so that the surrounding environment of the cells is a hypotonic solution, and at the same time assists in heating to achieve double effects. , causing the cells to absorb a large amount of water and swell, thereby strengthening the degree of cell lysis until the cell membrane finally ruptures, thereby realizing the c...

specific Embodiment approach 2

[0013] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the medium is DMEM high-glucose medium. Other steps and parameters are the same as those in the first embodiment.

specific Embodiment approach 3

[0014] Embodiment 3: This embodiment differs from Embodiment 1 or Embodiment 2 in that: the method for removing the culture medium is to use a pipette gun to absorb the culture medium, and wash it three times with isotonic PBS buffer. Other steps and parameters are the same as those in Embodiment 1 or 2.

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Abstract

The invention relates to a cell pretreatment method for electrochemical detection, which relates to a cell pretreatment method for electrochemical detection. The purpose of the present invention is to solve the problem that the electroactive substances released by the existing cell pretreatment technology are not complete enough, so that the change of cell activity cannot be truly reflected. Culture in an incubator with a volume concentration of 5%, then remove the medium, then add hypotonic lysate to adjust the cell concentration, and then lyse in a water bath to complete. The cell in-situ compound lysis of the present invention realizes the complete rupture of the cell membrane, greatly enhances the electrochemical signal, and simplifies the processing, which not only strengthens the electrochemical detection signal, but more importantly, the electrochemical detection result more accurately reflects the intracellular Reality of electroactive species content. The invention is applied in the field of biological analysis.

Description

technical field [0001] The invention relates to a cell pretreatment method for electrochemical detection. Background technique [0002] The in vitro cell electrochemical test method is a test method developed in recent years for the detection of cell viability. Compared with traditional methods, such as MTT method and flow cytometry method, it has the characteristics of simplicity, rapidity, sensitivity and low cost. , cell activity can be used as an indicator to evaluate the sensitivity of anticancer drugs, the toxicity of environmental carcinogens and the biocompatibility of biological materials. Therefore, the effective release of electroactive substances in cells is the key factor to complete the test, and the cell pretreatment techniques currently used to achieve the release of intracellular electroactive substances are cell heating lysis method and cell in situ hypotonic lysis method, namely Before the electrochemical detection, the cells are first placed in a device ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26
Inventor 崔继文毕胜野秀玉李锦莲刘继光武冬梅
Owner JIAMUSI UNIVERSITY
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