基于拉曼光谱方法研究不同剪切力下的红细胞亚损伤

doi:10.3964/j.issn.1000-0593(2021)06-1811-05

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Abstract In this study, confocal Raman spectroscopy was used to study the Sublethal damage erythrocytes of artificial heart pump under different shear stress to verify the ability of Raman spectroscopy to evaluate the degree of erythrocyte Sublethal damage. It provides a new idea for the evaluation of blood injury. The standard Raman spectra of hemoglobin and red blood cells were collected and compared in order to determine the peak intensity of erythrocytes. The sheer force of 0, 50, 100, 150, 200, 250 and 300 pa was applied to the tested blood samples for 1 s on the blood shear test platform. Using confocal Raman instrument, the Raman spectra of erythrocytes under shear stress were collected under 10 times long focal lens, 532 nm laser light source wavelength, integration time 10 s, twice, and 2.5 mW. Through the normalization method to compare the changes of the Raman spectrum of red blood cells, evaluate the degree of red blood cell Sublethal damage, and use the curve fitting method to fit the characteristic peak and shear stress to verify the ability of Raman spectrum to evaluate the sub-injury of red blood cells. Comparing the standard Raman spectra of hemoglobin and red blood cells, it is found that the erythrocyte spectra can reflect the internal structure of hemoglobin. The results show that Raman spectroscopy can be used to distinguish the Sublethal damage erythrocytes under different shear stress, and it is inferred that the shear stress can pass through the cell membrane and affect the internal hemoglobin structure. With the increase of shear force, the left spectral line of 1 376 cm^-1 increased. Obviously, the peak intensity of 1 549 and 1 604 cm^-1 increased, and the vibration band of oxygen concentration marker band 10 at 1 639 cm^-1 decreased. Among them, the peak strength of 1 549 cm^-1 position is the high spin band of ferrous ion. Under the action of different shear forces, the difference of peak strength is the most obvious, which has an obvious positive linear relationship with the shear stress, and the fitting effect is good. Raman spectroscopy has the advantages of simple treatment, short time, simple operation and good reproducibility can accurately detect the subtle changes in the internal structure of cells, and evaluate the degree of sub-injury of red blood cells. it makes up for the deficiency of the traditional evaluation of hemolysis, provides a new technical means for the evaluation of blood injury caused by artificial heart pump, and broadens the application field of the Raman detection method.

Key words：Red blood cell; Hemoglobin; Raman spectroscopy; Blood injury; Shear stress; Artificial heart

Received: 2020-11-17 Accepted: 2021-03-09

ZTFLH:

O657.37

Corresponding Authors: GAO Bin E-mail: gaobin@bjut.edu.cn

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https://www.gpxygpfx.com/EN/10.3964/j.issn.1000-0593(2021)06-1811-05 OR https://www.gpxygpfx.com/EN/Y2021/V41/I06/1811