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20 May 2022 Nonlinear optical microscopy of developing and brain tissue
Emmanuel Beaurepaire
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Proceedings Volume PC12136, Unconventional Optical Imaging III; PC1213601 (2022) https://doi.org/10.1117/12.2624475
Event: SPIE Photonics Europe, 2022, Strasbourg, France
Abstract
Many questions in developmental biology and neuroscience require tissue-scale measurements of multiple cell parameters. Recent progress in in-depth and large-scale microscopy approaches are transforming brain imaging, but generally lack efficient color contrast modalities. After introducing the basic concepts of multiphoton microscopy, the talk will discuss ongoing work aiming at augmenting the information content of multiphoton microscopy of tissues. First, we will present chromatic multiphoton serial (ChroMS) microscopy (1), a recent approach combining trichromatic two-photon (2P) excitation through wavelength mixing and microtome-based serial block-face image acquisition to acquire large-volume images of color-labelled brain tissue. This approach provides micrometric color imaging with constant resolution over the entire imaged volume. We illustrate its potential of ChroMS for several types of measurements, such as color-based morphological, clonal and connectivity analyses. Then, we will discuss three-photon (3P) microscopy, a recent approach providing superior imaging depth than 2P microscopy in highly scattering tissues, but requiring the development of novel infrared MHz OPA sources (2). Finally, we will discuss the combination of multiphoton fluorescence imaging with coherent contrast modalities providing complementary information such as third harmonic generation (THG) (3,4). References (1) Abdeladim et al, Nat Comm (2019) (2) Guesmi et al, Light Sci App (2018) (3) Morizet et al, Optica (2019) (4) Morizet et al, Optica (2021)
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Emmanuel Beaurepaire "Nonlinear optical microscopy of developing and brain tissue", Proc. SPIE PC12136, Unconventional Optical Imaging III, PC1213601 (20 May 2022); https://doi.org/10.1117/12.2624475
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KEYWORDS
Microscopy
Multiphoton microscopy
Brain
Optical microscopy
Tissue optics
Tissues
Coherence imaging
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