Study of the effect of alcohol on single human red blood cells using near-infrared laser tweezers Raman spectroscopy

Abstract

The effect of alcohol solution on single human red blood Cells (RBCs) was investigated using nearinfrared laser tweezers Raman spectroscopy (LTRS). In our system, a low-power diode laser at 785 nm was applied for the trapping of a living cell and the excitation of its Raman spectrum. Such a design could simultaneously reduce the photo-damage to the cell and suppress the interference from the fluorescence on the Raman signal. The denaturation process of single RBCs in 20% alcohol solution was investigated by detecting the time evolution of the Raman spectra at the single-cell level. The vitality of RBCs was characterized by the Raman band at 752 cm −1 , which corresponds to the porphyrin breathing mode. We found that the intensity of this band decreased by 34.1% over a period of 25 min after the administration of alcohol. In a further study of the dependence of denaturation on alcohol concentration, we discovered that the decrease in the intensity of the 752 cm −1 band became more rapid and more prominent as the alcohol concentration increased. The present LTRS technique may have several potential applications in cell biology and medicine, including probing dynamic cellular processes at the single cell level and diagnosing cell disorders in real time.

Raman spectroscopy: techniques and applications in the life sciences

“…The maximum value of a band is sometimes used as a simple substitute. Using the intensity of a single Raman band, Deng et al measured the effect of alcohol exposure on red blood cells [463] and Huang et al measured concentrations of citric acid in plasma [464]. Using multiple bands, Jung et al measured changes in DNA after cell adhesion [465] and Lee et al measured changes in DNA during apoptosis [466].…”

Sensitivity map of laser tweezers Raman spectroscopy for single-cell analysis of colorectal cancer

“…1,17 Briefly, the excitation beam of a diode laser ͑wavelength 782.5 nm͒ was delivered into the back pupil of the objective ͑100ϫ oil immersion͒ of a Nikon TE2000U dif-ferential interference contrast microscope. A laser trap was formed at the focal point.…”

Raman Spectroscopy of Blood and Blood Components

“…59 Laser tweezers Raman spectroscopy has become an increasingly important technique in the study of living RBCs, for investigations of oxygenation within single cells, [60][61][62] and for investigation of the effects of laser light on RBC biology. [63][64][65] The method has also been integral for characterizing red cell response to stimulus/ stress (e.g., alcohol-induced denaturation, 66 electrical current, 67 oxidative stress, 68 and pH changes 69 ), and has been deployed to compare oxygen uptake of different globin-containing cells. 70 Workers at the Center for Biophotonics, Science and Technology, UC Davis, who contributed to developing the LTRS capability, have studied a number of these phenomena.…”