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Broadband Coherent Anti-Stokes Raman Scattering (BCARS) Microscopy for High-Resolution Imaging of Dental Pulp Tissue.
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Edité par CCSD -
International audience. Dental pulp is a soft, complex tissue found inside the tooth root, made up of cells, blood vessels, and an extracellular matrix filled with organized fibers. This study explores the use of Broadband Coherent Anti-Stokes Raman Scattering (BCARS) microscopy to enhance chemical contrast, helping to better characterize dental pulp and analyze the unique spectral signatures of its individual cells. To obtain samples, pulp was extracted from the third molars of healthy patients. After removing the tooth crown, the pulp was carefully taken out and fixed in 4% paraformaldehyde (PFA). The samples were then embedded in paraffin, cut into ultra-thin 3 μm sections using a Microm HM340E with a Niagara system, and dewaxed with xylene to prepare for imaging. BCARS microscopy was performed using a Piezo X,Y,Z scanner (Nano-LP300, MCL), with imaging controlled through LabView software. Spectral data was extracted from raw BCARS signals using TDKK, a Python-based software (Crikit2, available on GitHub by C. Camp). The Raman spectral images covered a broad range (500–3800 cm⁻¹), capturing signals from weakly scattering modes and C-H vibrations. Spectral analysis revealed specific peaks linked to blood vessels (1651 cm⁻¹, 1440 cm⁻¹), as well as molecular signatures of lipids, proteins, phenylalanine, and nucleic acids in cells. Structural fibers such as elastin and collagen were also identified. These findings demonstrate that BCARS microscopy is a powerful tool for generating detailed chemical images of dental pulp, offering a deeper understanding of its composition and structure. By distinguishing various molecular components, this technique presents exciting possibilities for advanced tissue analysis in dentistry.
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