New Insight into Erythrocyte through In Vivo Surface-Enhanced Raman Spectroscopy

Abstract

The article presents a noninvasive approach to the study of erythrocyte properties by means of a comparative analysis of signals obtained by surface-enhanced Raman spectroscopy (SERS) and resonance Raman spectroscopy (RS). We report step-by-step the procedure for preparing experimental samples containing erythrocytes in their normal physiological environment in a mixture of colloid solution with silver nanoparticles and the procedure for the optimization of SERS conditions to achieve high signal enhancement without affecting the properties of living erythrocytes. By means of three independent techniques, we demonstrate that under the proposed conditions a colloid solution of silver nanoparticles does not affect the properties of erythrocytes. For the first time to our knowledge, we describe how to use the SERS-RS approach to study two populations of hemoglobin molecules inside an intact living erythrocyte: submembrane and cytosolic hemoglobin (Hb(sm) and Hb(c)). We show that the conformation of Hb(sm) differs from the conformation of Hb(c). This finding has an important application, as the comparative study of Hb(sm) and Hb(c) could be successfully used in biomedical research and diagnostic tests.

Building up spectral libraries for mapping erythrocytes by hyperspectral dark field microscopy

“…1 ((A) and (B), respectively). Three representative proteins were also chosen namely, hemoglobin and its heterocyclic organic portion of protoporphyrin IX, as well as spectrin, as cytoskeleton protein, thus providing some of the most relevant spectral characteristics related to membrane and submembrane environment of the RBC complementary to what has been described with other techniques, such as in vivo surface-enhanced Raman spectroscopy [1]. The spectra are shown in Fig.…”

Effect of hypoxia on the composition and state of lipids and oxygen-transport properties of erythrocyte haemoglobin

“…The conformation and properties of Hb were studied by Raman scattering (RS) using inVia confocal Raman microscope (Renishaw, Wotton-under-Edge, UK) [21][22][23][24] with short focus high luminosity monochromator (focal length not more than 250 mm). For excitation of Raman spectra, a laser was used (wavelength of 532 nm, maximum power of 100 mW and 100£ objective).…”

“…The position and intensity of the bands of the RS of Hb spectrum depends on the nature of fluctuations of the porphyrin ring bonds, which allows to evaluate the conformation of HP, which is directly related to the oxygen-binding properties of Hb (i.e. the Hb oxygen saturation) [23][24][25].…”

Violations of oxygen transport function of hemoglobin in surgical endointoxication and their correction using remaxol

“…Относительное ко-личество оксигемоглобина в эритроцитах опреде-ляли как отношение интенсивностей I 1375 /(I 1355 +I 1375 ) (I). Относительная способность гемоглобина свя-зывать лиганды рассчитывалась как отношение I 1355 /I 1550 (II), относительная способность гемогло-бина выделять лиганды -как отношение I 1375 /I 1580 (III), сродство гемоглобина к лигандам (О 2 ) -как отношение (I 1355 /I 1550 )/(I 1375 /I 1580 ) (IV), конформаци-онные изменения пирролов -как отношение I 1375 / I 1172 (V) [12,15]. При оценке тяжести перитонита ис-пользован Мангеймский индекс.…”