Cell pre-treatment method of electrochemical detection

An electrochemical and cellular technology, applied in the direction of electrochemical variables of materials, etc., can solve the problems of incomplete release of electroactive substances and inability to truly reflect changes in cell activity. The effect of the degree of lysis

Active Publication Date: 2015-11-04
JIAMUSI UNIVERSITY
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  • Summary
  • 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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  • Cell pre-treatment method of electrochemical detection
  • Cell pre-treatment method of electrochemical detection
  • Cell pre-treatment method of 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 pre-treatment method of electrochemical detection. In the prior art, in the existing cell pre-treatment technology, electrically active substances can not be completely released, such that the cell activity change can not be truly reflected. A purpose of the present invention is to solve the problem in the prior art. The method comprises: inoculating cells to be detected into a culture dish, placing into an incubator with a temperature of 37 DEG C and a CO2 volume concentration of 5% to culture, removing the culture medium, adding a hypotonic lysis liquid, adjusting the cell concentration, and carrying out lysis under a water bath condition so as to complete the process. According to the cell in situ composite lysis of the present invention, the complete rupture of the cell membrane is achieved, the electrochemical signal is substantially enhanced, and the treatment is simplified, such that the electrochemical detection signal is strengthened, and the importantly the electrochemical detection results accurately reflect the real situation of the intracellular electrically active substance content; and the cell pre-treatment method is used in the biological analysis field.

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 Applications(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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