Research Grants 23/04564-8 - Espectroscopia Raman, Biofísica - BV FAPESP
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PHENOTYPIC CHARACTERIZATION BY RAMAN MICROSCOPY, TERS AND SNOM IN SKELETAL MUSCLE FIBERS OF MICE

Abstract

The typification of skeletal muscle fibers in oxidative or glycolytic has great importance, both for the area of histology and for the area of real-time diagnostic biomarker, notably in the sports area. The classical phenotyping analyses are performed by immunofluorescence microscopy, using compatible primary and secondary antibodies, being a high technical challenge, since it involves high costs and great demand of time in sample preparation. Thus, the identification and characterization of new techniques for muscle cell typing with high accuracy and sensitivity, lower cost and time for sample preparation, have great potential for innovation. In this sense, the Raman microscopy technique may be useful for muscle cell phenotyping, as it allows the acquisition of a specific spectral signature for each type of muscle fiber, aided by microscopic images. For this purpose we will use cryopreserved soleus muscles taken from mice, fixed in paraformaldehyde (4%) and placed on glass slides. The Raman microscopy study will be performed in the spectral range of 500 to 3600 cm-1, using lasers with wavelengths of 473, 532, 633, 785 and 1064 nm to classify the fibers, type I, type II, type IIa, type IIx and type IIb, using primary and secondary antibodies. In addition, other sophisticated Raman spectroscopy techniques, such as TERS (tip enhanced Raman spectroscopy), will also be studied for the purpose of comparison with conventional Raman microscopy technique for muscle cell phenotyping. (AU)

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