Open Access Green: Postprint online available 12/2022 as soon as is submitted to ZB.
Wide-field mid-infrared hyperspectral imaging by snapshot phase contrast measurement of optothermal excitation.
Anal. Chem. 93, 15323-15330 (2021)
Vibrational microscopy methods based on Raman scattering or infrared absorption provide a label-free approach for chemical-contrast imaging, but employ point-by-point scanning and impose a compromise between the imaging speed and field-of-view (FOV). Optothermal microscopy has been proposed as a promising imaging modality to avoid this compromise, although at restrictively small FOVs capable of imaging only few cells. Here, we present wide-field optothermal mid-infrared microscopy (WOMiM) for wide-field chemical-contrast imaging based on snapshot pump-probe detection of optothermal signal, using a custom-made condenser-free phase contrast microscopy to capture the phase change of samples after mid-infrared irradiation. We achieved chemical contrast for FOVs up to 180 μm in diameter, yielding 10-fold larger imaging areas than the state-of-the-art, at imaging speeds of 1 ms/frame. The maximum possible imaging speed of WOMiM was determined by the relaxation time of optothermal heat, measured to be 32.8 μs in water, corresponding to a frame rate of μ30 kHz. This proof-of-concept demonstrates that vibrational imaging can be achieved at an unprecedented imaging speed and large FOV with the potential to significantly facilitate label-free imaging of cellular dynamics.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Lipid Bodies; Raman; Association; Transport; Fiber
ISSN (print) / ISBN 0003-2700
Journal Analytical Chemistry
Quellenangaben Volume: 93, Issue: 46, Pages: 15323-15330
Publisher American Chemical Society (ACS)
Publishing Place 1155 16th St, Nw, Washington, Dc 20036 Usa
Reviewing status Peer reviewed
Institute(s) Institute of Biological and Medical Imaging (IBMI)