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Method for detecting apoptosis of dinoflagellate cells

A dinoflagellate and cell technology, applied in the field of cell detection, can solve the problems of inability to distinguish dinoflagellate cells and the low accuracy of cell apoptosis detection results, and achieve the effects of quantitative reliability, accurate analysis and high result accuracy.

Active Publication Date: 2019-07-16
SHENZHEN UNIV
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Problems solved by technology

[0005] The purpose of the present invention is to overcome the described deficiency of prior art, provide a kind of method for detecting dinoflagellate cell apoptosis, to solve existing common flow cytometer can't distinguish dinoflagellate cell spontaneous Chla red fluorescence and the fluorescence that PI sends, As a result, the technical problem of low accuracy of cell apoptosis detection results

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  • Method for detecting apoptosis of dinoflagellate cells
  • Method for detecting apoptosis of dinoflagellate cells
  • Method for detecting apoptosis of dinoflagellate cells

Examples

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Embodiment 1

[0055] This example provides a method for detecting the apoptosis of dinoflagellate cells. The method for detecting the apoptosis of dinoflagellates comprises the following steps:

[0056] S11: Get 4 volumetric flasks and add dinoflagellate Tamar Alexandrina liquid respectively (the culture medium of Tamar Alexander dinoflagellate liquid is 2216E medium), divide described 4 volumetric flasks into 2 groups: experimental group (2 bottles), blank Control group (2 bottles); And make the dinoflagellate cell number of experimental group, contrast blank control group consistent; Wherein, add the 6A1 of volume 5% (wherein, 6A1 contains 10 8 cells / ml bacteriostatic cells, substratum is 2216E substratum); Add the 2216E substratum of the 6A1 of volume 5% in the volumetric flask of described control group (that is, the bacteriostatic cells that do not contain 6A1 in the control group); The experimental group and the control group were subjected to supernatant induction for 120min under t...

Embodiment 2

[0064] This example provides a method for detecting the apoptosis of dinoflagellate cells. The method for detecting the apoptosis of dinoflagellates comprises the following steps:

[0065] S11: Get 4 volumetric flasks and add dinoflagellate Tamar Alexandrina liquid respectively (the culture medium of Tamar Alexander dinoflagellate liquid is 2216E medium), divide described 4 volumetric flasks into 2 groups: experimental group (2 bottles), control group (2 bottles); and make the number of dinoflagellate cells of the experimental group and the control group consistent; wherein, add volume 10% 6A1 (wherein, 6A1 contains 10 8 cells / ml bacteriostatic cells, substratum is 2216E substratum); Add the 2216E substratum of the 6A1 of volume 10% in the volumetric flask of described control group (that is, the bacteriostatic cells that do not contain 6A1 in the control group); The experimental group and the control group were subjected to supernatant induction for 60 min under the same con...

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Abstract

The invention discloses a method for detecting apoptosis of dinoflagellate cells. The method for detecting the apoptosis of the dinoflagellate cells comprises the following steps: carrying out apoptosis processing on the dinoflagellate cells; carrying out concentration processing on the dinoflagellate cells subjected to the apoptosis processing; carrying out AnnexinV and propidine iodide (PI) dyeing processing on a concentrated dinoflagellate cell solution; carrying out imaging flow detection processing on a dyed dinoflagellate cell solution; and calculating an apoptosis rate of the dinoflagellate cells according to an AnnexinV / PI scatter diagram. The method for detecting the apoptosis of the dinoflagellate cells disclosed by the invention has the advantages that AnnexinV fluorescence, PIfluorescence and chlorophyll autofluorescence can be effectively distinguished through an imaging flow cytometer, so that the number of the dinoflagellate cells experiencing the apoptosis, which are among the dinoflagellate cells to be detected, can be directly and quantitatively measured, and an early cell apoptosis rate can be further obtained; and the quantification is reliable, the analysis isaccurate, and high-precision results can be obtained.

Description

technical field [0001] The invention belongs to the technical field of cell detection, and in particular relates to a method for detecting dinoflagellate cell apoptosis. Background technique [0002] Red tide is a harmful ecological phenomenon that causes the water body to change color after a large number of plankton blooms. There are more than 300 species that can cause red tides, such as the harmful dinoflagellate Alexandrium tamarense (Alexandriumtamarense), which can release paralytic shellfish poisoning ( paralyticshell fish poisoning, PSP), shellfish poisoning into the environment poses a serious threat to the marine economy and human health. Programmed cell death (PCD) is an autonomous and orderly death regulated by genes when cells grow or are subjected to environmental stress. In order to effectively control red tide, it is of great significance to understand the apoptosis of red tide dinoflagellates. [0003] At present, the widely used detection methods of cell ...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 陈辉蓉吴梓涵黎双飞胡章立陈秀锐李城彦
Owner SHENZHEN UNIV
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