„Quantitative Phase Imaging, from white light imaging to fluorescence super-resolution “

Quantitative Phase Imaging has been developed to retrieve information from biological samples. We have developed an approach based on generating self-interferences from the light emerging from an optical microscope [1]. More than a simple contrast enhancement technique, the quantitative information provided allows many possible applications, both in the coherent (transmission white-light) and the incoherent (fluorescence) world.
Initially developed for label-free microscopy, we have applied our approach to label-free live cells and tissue imaging. The sensitivity, its non-invasiveness as well as the versatility of this technique makes it applicable to multiple investigations such as label-free cytoskeleton imaging [2], identification of subcellular compounds [3] or dry mass measurement [4].
We have recently extended this idea to super-resolution by 3D super-localization of, first, absorbing (gold) nanoparticles [5] and then fluorescence single molecules [6]. The access to the phase of light unravels the axial position of a single emitter while the intensity gives the lateral position with neither PSF shaping nor multiple plane imaging. Our technique has the major advantage to work deep inside a tissue, allowing 3D super-resolution even at 25 µm without any adaptive optics.

References: [1] P. Bon, G. Maucort, B. Wattellier, and S. Monneret, „Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells,“ Opt. Express 17, (2009) 13080-13094 [2] P. Bon, S. Lécart, E. Fort and S. Lévêque-Fort „Fast label-free cytoskeleton network imaging in living mammalian cells”, Biophys. J. 106, (2014) 1588-1595 [3] P. Bon, J. Savatier, M. Merlin, B. Wattellier, S. Monneret, „Optical detection and measurement of living cell morphometric features with single-shot  uantitative phase microscopy,“ J. of Biomedical Optics, (2012), 17, 076004 [4] S. Aknoun, J. Savatier, P. Bon, F. Galland, L. Abdeladim, B. Wattellier, S. Monneret, “Living cell dry mass measurement using quantitative phase imaging with quadriwave lateral shearing interferometry: an accuracy and sensitivity discussion”, J. of Biomedical Optics, (2015), 20, 126009 [5] P. Bon, N. Bourg, S. Lécart, S. Monneret, E. Fort, J. Wenger, S. Lévêque-Fort “Three-dimensional nanometre localization of nanoparticles to enhance  super-resolution microscopy”, Nature Communications (2015), 6, 7764 [6] P. Bon et al, (2017)

Gastgeber:    

Microscopy Club of the Cluster of Excellence & DFG-Research Center
Nanoscale Microscopy and Molecular Physiology of the Brain
Alexander Egner &  Jörg Enderlein  & Stefan Jakobs
If you want to meet the speaker:  Please contact Tatjana Kasten (tatjana.kasten@llg-ev.de)

DFG Research Center CMPB  & Cluster of Excellence
Nanoscale Microscopy and Molecular Physiology of the Brain
Humboldtallee 23
37073 Göttingen
www.cmpb.de

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