A Method for Characterizing Absolute Intracellular Calcium Ion Concentrations Induced by Plasma

A technology of plasma and induced cells, applied in the preparation of test samples, material excitation analysis, fluorescence/phosphorescence, etc., to achieve wide application prospects, accurate measurement and characterization

Active Publication Date: 2020-09-22
DALIAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this characterization method measures the relative concentration of calcium ions
So far, no direct and accurate method for the determination of absolute calcium ion concentration has been reported

Method used

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  • A Method for Characterizing Absolute Intracellular Calcium Ion Concentrations Induced by Plasma
  • A Method for Characterizing Absolute Intracellular Calcium Ion Concentrations Induced by Plasma
  • A Method for Characterizing Absolute Intracellular Calcium Ion Concentrations Induced by Plasma

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Embodiment 1

[0038] A method for characterizing the absolute intracellular calcium ion concentration induced by plasma comprising the steps of:

[0039] Step 1, Prepare Cell Suspension

[0040] Saccharomyces cerevisiae CGMCC no.6148 was cultured in YPD culture medium (10g / L yeast extract powder, 20g / L peptone, 20g / L glucose) at 28°C, 170rpm; collect Saccharomyces cerevisiae cells in logarithmic growth phase, centrifuge at 4°C, 30000 ×g, 5min, discard the supernatant, resuspend in fresh 2% YPD culture medium, adjust the bacterial concentration OD 600 to 6.0, when the cell concentration is 5×10 5 individual / mL.

[0041] Step 2, Optimizing Fluo-3 AM Use Conditions

[0042] (1) Optimization of water bath temperature and incubation time

[0043] Divide the cell suspension prepared in step 1 into 1.5mL centrifuge tubes, 1mL in each tube, 6 tubes in total, and each 6 tubes is a group, and each group is treated in a water bath at 30°C, 37°C, and 44°C for 0 min , 20min, 40min, 60min, 80min, 10...

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Abstract

The invention relates to a method for representing plasma induced intracellular absolute calcium ion concentration. According to the method, a prepared cell suspension is added into an optimized fluo-3AM, 0.8 mM Pluronic F-127 is added, and a maximum fluorescence intensity Fmax value is obtained, or a calcium ion chelating agent EGTA is added, and a minimum fluorescence intensity Fmin value is obtained; the prepared cell suspension is added into the optimized fluo-3AM and then subjected to atmospheric pressure cold plasma treatment, and by combining with a multifunctional microplate reader todetermine an F value, the absolute calcium ion concentration is calculated according to a formula. The method for measuring the absolute calcium ion concentration of cells by combining the fluorescentprobe flou-3AM with the multifunctional microplate reader is achieved and has the advantages of being sensitive, highly efficient, simple and accurate in determination and representing the intracellular absolute calcium ion concentration.

Description

technical field [0001] The invention belongs to the technical field of quantitative detection of intracellular calcium ion concentration, in particular to a method for characterizing the absolute intracellular calcium ion concentration induced by plasma. Background technique [0002] As an important second messenger, calcium ions participate in various signal transduction in cells and regulate a series of life activities. Intracellular calcium ion concentration within a certain range (1 ~ 10μM) is conducive to microbial growth and product formation, but too high calcium ion concentration will inhibit the growth of bacteria, and even promote the death of bacteria. Atmospheric pressure-cooled plasma, as an enhanced technology, can enhance the ability of microorganisms to transform target products by inducing changes in the concentration of calcium ions in cells. At present, there are many limiting factors in the commonly used methods for the determination of intracellular cal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N1/44
CPCG01N1/44G01N21/6402G01N21/6486
Inventor 董晓宇
Owner DALIAN UNIV
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