Dynamic molecular monitoring of retina inflammation byin vivoRaman spectroscopy coupled with multivariate analysis

Abstract

Retinal tissue is damaged during inflammation in Multiple Sclerosis. We assessed molecular changes in inflamed murine retinal cultures by Raman spectroscopy. Partial Least Squares‐Discriminant analysis (PLS‐DA) was able to classify retina cultures as inflamed with high accuracy. Using Multivariate Curve Resolution (MCR) analysis, we deconvolved 6 molecular components suffering dynamic changes along inflammatory process. Those include the increase of immune mediators (Lipoxygenase, iNOS and TNFα), changes in molecules involved in energy production (Cytochrome C, phenylalanine and NADH/NAD+) and decrease of Phosphatidylcholine. Raman spectroscopy combined with multivariate analysis allows monitoring the evolution of retina inflammation.Raman spectroscopy analysis of the Retinal Ganglion Cell layer of the retina. (A) Design of the analysis of the Ganglion cell layer (GCL) and Retinal Nerve Fiber Layer (RNFL) of the retina based in the physical properties of laser light and anatomical structure of retinal layers. (B) Examples of raw Raman spectra from representative retina sample after 10 hours incubation time (black) and LPS treated retina sample after 10 hours incubation time (red) and 12 hours incubation time (blue). (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Combining hyperspectral imaging and chemometrics to assess and interpret the effects of environmental stressors on zebrafish eye images at tissue level

“…The so‐called fingerprint region of Raman spectra (900‐1800 cm −1 ) is usually acquired for biological samples because it contains wide information about different families of biological molecules , for example, lipids, proteins, DNA and so forth. Raman spectroscopy coupled with multivariate curve resolution (MCR) analysis has been used to monitor different biological molecular components in biochemical processes in tissues or cells .…”

Biological and Medical Applications of Multivariate Curve Resolution Assisted Raman Spectroscopy

“…93 Dynamic changes of molecular components such as increase of immune mediators, changes in energy producing molecules and decrease of phosphatidylcholine were observed during the evolution of retina inflammation which may become a promising application to study retina disease in vivo.…”

Rapid spontaneous Raman light sheet microscopy using cw-lasers and tunable filters

“…Although the signal of typical CMOS sensitivity is compromised due to the low quantum efficiency at the scattered Raman wavelengths (800-900 nm) it has been demonstrated that no damage is induced in cells when Raman experiments are conducted during 40 minutes of exposure using a 785 nm source at 115 mW with a 63x 0.9 NA objective [26]. An alternative way to extract the Raman molecular information from the low resolved spectra is applying modern deconvolution methods based on, for example, multivariate curve resolution (MCR) [27], which has been used to resolve multiple pure responses and concentrations of the Raman components present in unknown mixtures [28]. Here we are working in the CH stretching region where many molecular components share bands or are highly overlapped, thus the combination of the Raman LiShMS imaging technique here presented with MCR would enormously enhance the results and the chemical knowledge of the sample under analysis.…”