In vivo hyperspectral CARS and FWM microscopy of carotenoid accumulation in H. pluvialis

  1. Get@NRC: In vivo hyperspectral CARS and FWM microscopy of carotenoid accumulation in H. pluvialis (Opens in a new window)
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Proceedings titleProgress in Biomedical Optics and Imaging - Proceedings of SPIE
ConferenceMultimodal Biomedical Imaging IX, 1 February 2014 through 2 February 2014, San Francisco, CA
Article number893709
SubjectAstaxanthin; Carotenogenesis; Coherent anti-Stokes Raman scattering; Haematococcus pluvialis; Nonlinear optical microscopy; Algae; Chlorophyll; Fluorescence; Medical imaging; Optical data storage; Optical microscopy; Pigments; Raman scattering; Raman spectroscopy; Coherent scattering
AbstractCoherent anti-Stokes Raman scattering (CARS) and four-wave-mixing (FWM) microscopy are a related pair of powerful nonlinear optical characterization tools. These techniques often yield strong signals from concentrated samples, but because of their quadratic dependence on concentration, they are not typically employed for imaging or identifying dilute cellular constituents. We report here that, depending on the excitation wavelengths employed, both CARS and degenerate-FWM signals from carotenoid accumulations in alga cysts can be exceptionally large, allowing for low-power imaging of astaxanthin (AXN) deposits in Haematococcus pluvialis microalga. By use of a broadband laser pulse scheme for CARS and FWM, we are able to simultaneously collect strong intrinsic two-photon-excitation fluorescence signals from cellular chlorophyll in vivo. We show that CARS signals from astaxanthin (AXN) samples in vitro strictly follow the expected quadratic dependence on concentration, and we demonstrate the collection of wellresolved CARS spectra in the fingerprint region with sensitivity below 2mM. We suggest that multimodal nonlinear optical microscopy is sufficiently sensitive to AXN and chlorophyll concentrations that it will allow for non-invasive monitoring of carotenogenesis in live H. pluvialis microalgae.
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AffiliationNational Research Council Canada; Security and Disruptive Technologies; Aquatic and Crop Resource Development
Peer reviewedYes
NPARC number21272669
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Record identifierd22efeba-5dbd-4c70-80a6-fe254c4f6b13
Record created2014-12-03
Record modified2016-05-09
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