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Method for detecting pH value in red blood cells by using autofluorescence of red blood cells

An autofluorescence and red blood cell technology, applied in the field of analytical chemistry, can solve the problems of being easily affected and large detection errors, and achieve the effects of short time-consuming, high precision and simple operation

Pending Publication Date: 2021-06-08
ZHONGSHAN HOSPITAL FUDAN UNIV
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Problems solved by technology

However, the existing common intracellular pH detection method mainly adopts incubating pH-sensitive ratiometric nanoprobes, but red blood cells have autofluorescence effect, which is easily affected when it is applied to the detection of red blood cell intracellular pH, and the detection error is large.

Method used

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  • Method for detecting pH value in red blood cells by using autofluorescence of red blood cells
  • Method for detecting pH value in red blood cells by using autofluorescence of red blood cells
  • Method for detecting pH value in red blood cells by using autofluorescence of red blood cells

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Embodiment

[0036] Embodiment: A kind of method utilizing red blood cell autofluorescence to detect the pH value in red blood cell, it comprises the following steps:

[0037] S1: Take 50 μl of whole blood, centrifuge at 1500 rpm for 5 minutes, and discard the supernatant to obtain red blood cells;

[0038] S2: Wash the erythrocytes prepared in step S1 with PBS, centrifuge at 1500 rpm for 5 min, and discard the supernatant;

[0039] S3: Add PBS to resuspend the suspension of S2 after discarding the supernatant, and then divide the same amount into several EP tubes;

[0040] S4: Centrifuge the liquid in the EP tube at 300×g for 10 min, discard the supernatant;

[0041] S5: Add calibration solutions and sample buffers with different pH values ​​into the EP tube, incubate at 37°C for 15 minutes, and prepare red blood cell calibration solution samples with different pH values; characterize the distribution of red blood cells in the calibration samples, and the characterization results See ...

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Abstract

The invention discloses a method for detecting pH value in red blood cells by using autofluorescence of red blood cells, which comprises the following steps: preparing calibration solutions with different pH values, collecting fluorescence intensity signals of the calibration solutions, drawing a standard curve of average fluorescence intensity and pH to obtain a linear equation, and substituting the average fluorescence intensity in a red blood cell sample to be detected into the standard curve. The pH value in the red blood cells to be detected can be obtained. The method for detecting the pH value in red blood cells has the advantages of simplicity in operation, short time consumption, stable detection result and the like. According to the method, the autofluorescence characteristic of the red blood cells is utilized, fluorescent dye does not need to be additionally added or prepared, a standard curve of the change of the fluorescence intensity of the red blood cells in different pH environments is established, and the corresponding pH values in the red blood cells with different fluorescence intensities are calculated in turn. The accuracy value of the pH value of the method can reach 0.01 pH unit, and the accuracy is high.

Description

technical field [0001] The invention relates to the technical field of analytical chemistry, in particular to a method for detecting pH value in red blood cells by using red blood cell autofluorescence. Background technique [0002] Body fluid accounts for 60%-70% of the human body, 2 / 3 of which are located in the cells. Acidity and alkalinity (pH), especially the pH of intracellular fluid, plays an important regulatory role in various cell biological behaviors, such as enzyme activity, cell proliferation and apoptosis, information transmission and tumor cell drug resistance, etc., and is an important microenvironment that affects cell function index. [0003] However, the current clinical blood gas analysis technology measures the pH in serum, which is equivalent to the pH of extracellular fluid, and the acid-base change of intracellular fluid, which accounts for the vast majority of body fluids, and the relationship between the acid-base balance of intracellular and extra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N15/14G01N1/28G01N1/34G01N1/40
CPCG01N21/6486G01N15/14G01N1/28G01N1/34G01N1/40G01N1/4077
Inventor 丁小强宋娜娜张健陆雨菲
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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