Picosecond resonance Raman spectrum of the oxyhemoglobin photoproduct. Evidence for an electronically excited state

Terner, James; Voss, David; Paddock, Carolyn A.; Miles, Richard B.; Spiro, Thomas G.

doi:10.1021/j100395a001

Cited by 22 publications

(19 citation statements)

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“…The respective Soret maxima of oxy-and deoxy-Hb occur at 415 and 430 nm, and excitation within the Soret band of heme proteins provides strong enhancement in the 1,350-to 1,380-cm Ϫ1 range, also known as 4 (1, 36). While the oxidation state marker band 4 is the highest intensity heme Raman band under Soret excitation, the 19 and 10 bands (1,500 -1,650 cm Ϫ1 range) are usually more prominent under 532-nm excitation (35,37,39).The determination of Hb O 2 saturation (SO 2 ) using Raman spectroscopy has the advantage that the oxygenation information is only related to the specific interaction of the photons, with the Hb molecules generating a unique Raman spectrum (4,5,9,34,38,39). Our laboratory has recently reported that this technology can be adapted to the measurement of SO 2 in flowing blood of single microvessels of living tissues, in a configuration that provides the measurement of other microvascular parameters, like diameter and blood flow, using other noninvasive optical techniques (42).…”

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“…The determination of Hb O 2 saturation (SO 2 ) using Raman spectroscopy has the advantage that the oxygenation information is only related to the specific interaction of the photons, with the Hb molecules generating a unique Raman spectrum (4,5,9,34,38,39). Our laboratory has recently reported that this technology can be adapted to the measurement of SO 2 in flowing blood of single microvessels of living tissues, in a configuration that provides the measurement of other microvascular parameters, like diameter and blood flow, using other noninvasive optical techniques (42).…”

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confidence: 99%

“…The availability of components for wavelengths such as 532 nm is presently less limited than for 406 nm, and most commercially available systems employ 532-nm lasers. In addition, Raman spectra with 532-nm excitation are also well known and distinctive for oxy-and deoxy-Hb (39). Finally, the penetration of 532-nm excitation is deeper than that using a 406-nm laser, allowing a larger fraction of a blood vessel (or vessels located deeper within the tissue) to be probed for SO 2 levels.…”

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Measurement of hemoglobin oxygen saturation using Raman microspectroscopy and 532-nm excitation

Filho¹,

Terner²,

Pittman³

et al. 2008

Journal of Applied Physiology

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The resonant Raman enhancement of hemoglobin (Hb) in the Q band region allows simultaneous identification of oxy- and deoxy-Hb. The heme vibrational bands are well known at 532 nm, but the technique has never been used to determine microvascular Hb oxygen saturation (So(2)) in vivo. We implemented a system for in vivo noninvasive measurements of So(2). A laser light was focused onto areas of 15-30 microm in diameter. Using a microscope coupled to a spectrometer and a cooled detector, Raman spectra were obtained in backscattering geometry. Calibration was performed in vitro using blood at several Hb concentrations, equilibrated at various oxygen tensions. So(2) was estimated by measuring the intensity of Raman signals (peaks) in the 1,355- to 1,380-cm(-1) range (oxidation state marker band nu(4)), as well as from the nu(19) and nu(10) bands (1,500- to 1,650-cm(-1) range). In vivo observations were made in microvessels of anesthetized rats. Glass capillary path length and Hb concentration did not affect So(2) estimations from Raman spectra. The Hb Raman peaks observed in blood were consistent with earlier Raman studies using Hb solutions and isolated cells. The correlation between Raman-based So(2) estimations and So(2) measured by CO-oximetry was highly significant for nu(4), nu(10), and nu(19) bands. The method allowed So(2) determinations in all microvessel types, while diameter and erythrocyte velocity could be measured in the same vessels. Raman microspectroscopy has advantages over other techniques by providing noninvasive and reliable in vivo So(2) determinations in thin tissues, as well as in solid organs and tissues in which transillumination is not possible.

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confidence: 99%

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Measurement of hemoglobin oxygen saturation using Raman microspectroscopy and 532-nm excitation

Filho¹,

Terner²,

Pittman³

et al. 2008

Journal of Applied Physiology

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“…In terms of HbO 2 Sat determination, this method has the advantage that the oxygenation information is related only to the specific interaction of the photons with the Hb molecules, generating a unique Raman spectrum (6,7,12,36,40,41). The vibrational bands of heme are well known (1,5), and previous Raman investigations of Hb have been carried out on isolated and purified Hb or in isolated red blood cells (27,33,34).…”

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Hemoglobin oxygen saturation measurements using resonance Raman intravital microscopy

Filho

Terner

Pittman

et al. 2005

American Journal of Physiology-Heart and Circulatory Physiology

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A system is described for in vivo noninvasive measurements of hemoglobin oxygen saturation (HbO2Sat) at the microscopic level. The spectroscopic basis for the application is resonant Raman enhancement of Hb in the violet/ultraviolet region, allowing simultaneous identification of oxy- and deoxyhemoglobin with the same excitation wavelength. The heme vibrational bands are well known, but the technique has never been used to determine microvascular HbO2Sat in vivo. A diode laser light (power: 0.3 mW) was focused onto sample areas 15–30 μm in diameter. Raman spectra were obtained in backscattering geometry by using a microscope coupled to a spectrometer and a cooled detector. Calibration was performed in vitro by using glass capillaries containing blood at several Hb concentrations, equilibrated at various oxygen tensions. HbO2Sat was estimated using the Raman band intensities at 1,360 and 1,375 cm−1. Glass capillary path length and Hb concentration had no effect on HbO2Sat estimated from Raman spectra. In vivo observations were made in blood flowing in microvessels of the rat mesentery. The Hb Raman peaks observed in oxygenated and deoxygenated blood were consistent with earlier Raman studies that used Hb solutions and isolated cells. The method allowed HbO2Sat determinations in the whole range of arterioles, venules, and capillaries. Tissue transillumination allowed diameter and erythrocyte velocity measurements in the same vessels. Raman microspectroscopy offers distinct advantages over other currently used techniques by providing noninvasive and reliable in vivo determinations of HbO2Sat in thin tissues as well as in solid organs and tissues, which are unsuitable for techniques requiring transillumination.

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Applications in Bioinorganic Chemistry

Nakamoto

2008

Infrared and Raman Spectra of Inorganic and Coordination Compounds

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Picosecond resonance Raman spectrum of the oxyhemoglobin photoproduct. Evidence for an electronically excited state

Cited by 22 publications

References 1 publication

Measurement of hemoglobin oxygen saturation using Raman microspectroscopy and 532-nm excitation

Measurement of hemoglobin oxygen saturation using Raman microspectroscopy and 532-nm excitation

Hemoglobin oxygen saturation measurements using resonance Raman intravital microscopy

Applications in Bioinorganic Chemistry

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