Raman spectroscopic investigations on optical trap induced deoxygenation of red blood cells

Ahlawat, Sunita; Kumar, Nitin; Dasgupta, Ranjan; Verma, Ravi Shanker; Uppal, Abha; Gupta, Pradeep Kumar

doi:10.1063/1.4828706

Cited by 23 publications

(21 citation statements)

References 22 publications

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“…[28] The peaks present in spin marker region (1,500-1,650 cm -1 ) is originating from the C-C bonds in the porphyrin ring, which in turn is influenced by the spin state of central Fe atom. [29] As mentioned previously, the iron atom at the center of porphyrin ring is pulled apart during oxy-deoxy transition, which results in the decrease in coordination strength between it and the ring. This is sufficient to impart variation in the molecular vibrations.…”

Section: Resultsmentioning

confidence: 88%

“…Due to the close proximity of this vibrations to protein subunits, any conformational alterations occurring inside hemoglobin can be immediately tracked from the Raman oxygenation signatures present in this region . The peaks present in spin marker region (1,500–1,650 cm ‐1 ) is originating from the CC bonds in the porphyrin ring, which in turn is influenced by the spin state of central Fe atom . As mentioned previously, the iron atom at the center of porphyrin ring is pulled apart during oxy‐deoxy transition, which results in the decrease in coordination strength between it and the ring.…”

Section: Resultsmentioning

confidence: 99%

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Optical tweezers combined with micro‐Raman investigation of alcohol‐induced changes on single, live red blood cells in blood plasma

Lukose

Mithun

Mohan

et al. 2019

J Raman Spectroscopy

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Alcohol is commonly used as sterilizing agent in hospital settings. Alcohol abuse is also associated with numerous medical complications. It influences the coagulation mechanics and erythropoiesis. Optical tweezers are being extensively used in research labs for the trapping as well as manipulation of biological cells. The biochemical investigation of trapped biological cells can be realized with the integration of Raman spectroscopic technique with optical tweezers, which is termed as Raman tweezers. In this work, we interrogate the influence of ethanol exposure on human red blood cells at single cell level using micro‐Raman spectroscopy. In order to mimic in vivo condition to an extent, the experiment is performed by optically trapping the live cell in blood plasma. In the present work, a 785‐nm diode laser was used for trapping and probing of single, live red blood cell. An intensity deduction was noted for porphyrin breathing mode at 752 cm‐1 in the Raman spectra as a function of ethanol concentration, which indicates the depletion in hemoglobin. Hemoglobin deoxygenation process in the presence of alcohol is also evident from the variations in major oxygenation marker bands at 1,209; 1,222; 1,544; 1,561 cm‐1, etc., in the Raman spectrum of red blood cell. The effect of alcohol on red blood cells in blood plasma induces membrane depletion and hemoglobin degradation. This is also accompanied by the conversion of oxygenated hemoglobin to more deoxygenated state. Present study has also demonstrated the capability of micro‐Raman spectroscopy as a promising tool for probing red blood cell oxygenation status.

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Section: Resultsmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Optical tweezers combined with micro‐Raman investigation of alcohol‐induced changes on single, live red blood cells in blood plasma

Lukose

Mithun

Mohan

et al. 2019

J Raman Spectroscopy

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“…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.…”

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confidence: 99%

Raman Spectroscopy of Blood and Blood Components

et al. 2017

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Blood is a bodily fluid that is vital for a number of life functions in animals. To a first approximation, blood is a mildly alkaline aqueous fluid (plasma) in which a large number of free-floating red cells (erythrocytes), white cells (leucocytes), and platelets are suspended. The primary function of blood is to transport oxygen from the lungs to all the cells of the body and move carbon dioxide in the return direction after it is produced by the cells' metabolism. Blood also carries nutrients to the cells and brings waste products to the liver and kidneys. Measured levels of oxygen, nutrients, waste, and electrolytes in blood are often used for clinical assessment of human health. Raman spectroscopy is a nondestructive analytical technique that uses the inelastic scattering of light to provide information on chemical composition, and hence has a potential role in this clinical assessment process. Raman spectroscopic probing of blood components and of whole blood has been on-going for more than four decades and has proven useful in applications ranging from the understanding of hemoglobin oxygenation, to the discrimination of cancerous cells from healthy lymphocytes, and the forensic investigation of crime scenes. In this paper, we review the literature in the field, collate the published Raman spectroscopy studies of erythrocytes, leucocytes, platelets, plasma, and whole blood, and attempt to draw general conclusions on the state of the field.

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“…4(e) and 4(f) which displays the mean of (n = 14 time series) difference spectra between the mean of the first 10 spectra and the last 10 spectra for each time series, along with 95% confidence interval, for static and flowed blood respectively. The longer laser exposure time of 200 s for the static blood volume could be causing photoinduced oxidation in the static blood as shown in previous reports [21,[36][37][38]. In contrast each flowed blood volume is exposed to the focused laser spot for only 0.4 s at the 100 µm depth inside the quartz flow cell.…”

Section: Comparison Of the Raman Spectra Of Flowed And Static Bloodmentioning

confidence: 80%

Development of a flow cell based Raman spectroscopy technique to overcome photodegradation in human blood

Hansson

Allen

Qutob

et al. 2019

Biomed. Opt. Express

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Raman spectroscopy of blood offers significant potential for label-free diagnostics of disease. However, current techniques are limited by the use of low laser power to avoid photodegradation of blood; this translates to a low signal to noise ratio in the Raman spectra. We developed a novel flow cell based Raman spectroscopy technique that provides reproducible Raman spectra with a high signal to noise ratio and low data acquisition time while ensuring a short dwell time in the laser spot to avoid photodamage in blood lysates. We show that our novel setup is capable of detecting minute changes in blood lysate spectral features from natural aging. Moreover, we demonstrate that by rigorously controlling the experimental conditions, the aging effect due to natural oxidation does not confound the Raman spectral measurements and that blood treated with hydrogen peroxide to induce oxidative stress can be discriminated from normal blood with a high accuracy of greater than 90% demonstrating potential for use in a clinical setting.

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Raman spectroscopic investigations on optical trap induced deoxygenation of red blood cells

Cited by 23 publications

References 22 publications

Optical tweezers combined with micro‐Raman investigation of alcohol‐induced changes on single, live red blood cells in blood plasma

Optical tweezers combined with micro‐Raman investigation of alcohol‐induced changes on single, live red blood cells in blood plasma

Raman Spectroscopy of Blood and Blood Components

Development of a flow cell based Raman spectroscopy technique to overcome photodegradation in human blood

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