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Resonance Raman spectroscopy of human sickle cell hemoglobin from transgenic mice

We have previously developed a methodology to measure hemoglobin (Hb) oxygen saturation using Raman spectroscopy in vivo (AJP 289:H488, 2005; JAP 104:1809, 2008). However, the technique has not yet been used in sickle cell disease (SCD). We have established a colony of transgenic (Tg) SCD mice. This strain no longer expresses mouse Hba and Hbb, but express human sickle hemoglobin (HbS). It mimics the genetic, hematologic and histopathologic features found in humans with SCD. Whole blood samples were obtained from anesthetized Tg mice and normal control mice. Raman spectra from these samples were obtained in vitro using the exciting wavelengths in the range of 405 ‐ 532 nm emitted by a tunable laser. The scattered light was sampled at 90 degrees using a 0.5 m spectrometer and a nitrogen cooled CCD detector. In order to induce HbS polymerization, Raman spectra were collected from blood samples exposed to high and low oxygen environments. The findings suggest that strong Raman signals can be obtained from normal and SCD blood, opening the possibility for in vivo noninvasive measurements of hemoglobin Hb and HbS saturation in the microcirculation. The resonance Raman technique should provide additional worthwhile microscopic information in the study of the abnormal hemoglobins in vivo.