Raman spectroscopy is a highly sensitive and specific technique for identifying tissue compositions. Raman-based characterization of normal and abnormal tissues is impeded due to the variability in routine tissue preparation techniques, fluorescent background, and molecular heterogeneity. Thus, sample preparation and Raman measurement conditions for tissue sections must be optimized. Here, we present an optimized Raman protocol and sample preparation method for brain tissue sections. This protocol allows the characterization of tissues and recognition of brain tumors by refining laser power, accumulation/exposure times, excitation wavelength, glass/CaF2 substrate, deparaffinization solvent, and the thickness of sections.
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