Quantitative phase microscopy for Biology

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using wavefront imaging

Principal investigators : Serge Monneret, Guillaume Baffou, Julien Savatier

keywords : lateral shearing interferometry, wavefront microscopy

Quantitative phase imaging (QPI) techniques are now conventionally used in microscopy for measuring specific properties of semi-transparent samples without any labelling. By placing a wavefront imaging system on the exit image plane of a microscope, we measure the complex field spatial distribution in this plane, and then retrieve the quantitative optical path difference (OPD) of the observed sample.

The QPI technique we use is a wavefront imaging technique based on Quadriwave Lateral Shearing Interferometry (QLSI). It consists of a 2D-grating placed at a millimetric distance from a regular camera. QLSI can reach the diffraction limit, it is achromatic, and thus compatible with the white-light illumination pathway of conventional wide-field microscopes.

In the last years, we extended the technique to new applications, where different physical phenomena produced a given sample-induced change in the phase of the exit optical beam that modulates the incident wavefront. More precisely, we used direct refractive-induced, thermal-induced, and resonant vibrational-induced OPD variations to study physical or biological samples, the retardance, temperature distribution in living cells, the optical properties of nanoparticles and 2D-materials, and the dry mass of bacteria and neurons in culture.

Application area

Quantitative phase microscopy on living cells, optical waveguides, laser damage sites
Dry mass determination of cells / Cell cycle follow-up
Temperature microscopy
Quantitative phase imaging with nonlinear microscopy (CARS)
Refractive index and thermo-refractive index (dn/dT) measurement of liquids
Phase tomography
Optical retardance imaging / specific imaging of birefringent samples
3D localization of metallic nanoparticles
Optical profilometry on etched surfaces, laser damage craters


Single-shot quantitative phase-fluorescence imaging using cross-grating wavefront microscopy
M. Bénéfice, A. Gorlas, B. Marthy, V. Da Cunha, P. Forterre, A. Sentenac, P. C. Chaumet, G. Baffou
Scientific Report 14, 2142 (2024)
Dry mass photometry of single bacteria using quantitative wavefront microscopy
M. Bénéfice, A. Gorlas, B. Marthy, V. Da Cunha, P. Forterre, A. Sentenac, P. C. Chaumet, G. Baffou
Biophysical Journal 122, 1-14 (2023)
Biomass measurements of single neurites in vitro using optical wavefront microscopy
L. Durdevic, A. Relaño Ginés, A. Roueff, G. Blivet, G. Baffou
Biomedical Optics Express 13, 6550 (2022).
Life at high temperature observed in vitro upon laser heating of gold nanoparticles
C. Molinaro, M. Bénéfice, A. Gorlas, V. Da Cunha, H. M. L. Robert, R. Catchpole, L. Gallais, P. Forterre, G. Baffou
Nature Communications 13, 5342 (2022).
Quantitative phase imaging of adherent mammalian cells : a comparative study
Allier, C ; Herve, L ; (...) ; Morales, S
Biomed. Opt. Exp. 10, 2768-2783 (2019).
Wavefront sensing applied to determine the temperature dependence of the refractive index of liquids
Radhakrishnan, R ; Gallais, L and Monneret, S
Appl. Opt. 58, 3646-3651 (2019).
Large field-of-view phase and fluorescence mesoscope with microscopic resolution
de Kernier, I ; Ali-Cherif, A ; (...) ; Blandin, P ; Rongeat, N. ; Cioni, O. ; Morales, S. ; Savatier, S. ; Monneret, S. ; Blandin, P.
J. Biomed. Opt. 24, 036501 (2019).
Quantitative retardance imaging by means of quadri-wave lateral shearing interferometry for label-free fiber imaging in tissues
Aknoun, S ; Aurrand-Lions, M ; Wattellier, B. ; Monneret, S
Opt. Commun. 422, 17-27 (2018).
Laser-induced birefringence measurements by quantitative polarized-phase microscopy
Doualle, T ; Olle, A ; Cormont, P. ; Monneret, S. ; Gallais, L
Optics Letters 42(8), 1616-1619 (2017).
Time-resolved quantitative phase microscopy of laser-material interactions using a wavefront sensor
L. Gallais, S. Monneret
Optics Letters 41(14), 3245-3248 (2016).
Living cells dry mass measurements using Quantitative Phase Imaging with Quadri Wave Lateral Shearing Interferometry. An accuracy and sensitivity discussion
S. Aknoun, J. Savatier, P. Bon, F. Galland, L. Abdeladim, B. Wattellier, S. Monneret
Journal of Biomedical Optics 20(12), 126009 (2015).
Quantitative phase imaging applied to laser damage detection and analysis
D.-B. Douti, M. Chrayteh, S. Aknoun, T. Doualle, C. Hecquet, S. Monneret, L. Gallais
Applied Optics 28 (54), 8375-8382 (2015)
Three dimensional nanometer localization of nanoparticles to stabilize super-resolution microscopy
P. Bon, N. Bourg, S. Lécart, S. Monneret, E. Fort, J. Wenger, S. Lévêque-Fort
Nature Communications 6, 7764 (2015)
Quantitative Retardance Imaging of biological samples using Quadri-Wave Lateral Shearing Interferometry
S. Aknoun, P. Bon, J. Savatier, B. Wattellier, S. Monneret
Optics Express 23 (12), 16383-16406 (2015).
Photo-induced heating of nanoparticle arrays
G. Baffou, P. Berto, E. Bermudez, R. Quidant, S. Monneret, J. Polleux, H. Rigneault
ACS Nano 7 (11), 6478-6488 (2013)
Three-dimensional temperature imaging around a gold microwire
P. Bon, N. Belaid, D. Lagrange, C. Bergaud, H. Rigneault, S. Monneret, G. Baffou
Applied Physics Letters 102, 244103 (2013)
Wide-field vibrational phase imaging in an extremely folded box-coherent anti-Stokes Raman scattering geometry
P. Berto, A. Jesacher, C. Roider, S. Monneret, H. Rigneault, M. Ritsch-Marte
Optics Letters 38 (5), 709-711 (2013).
Wide-field vibrational phase imaging
P. Berto, D. Gachet, P. Bon, S. Monneret, H. Rigneault
Physical Review Letters 109, 093902 (2012).
Imaging the Gouy phase shift in photonic jets with a wavefront sensor
P. Bon, B. Rolly, N. Bonod, J. Wenger, B. Stout, S. Monneret, H. Rigneault
Optics Letters 37 (17), 3531-3533 (2012).
Non-iterative boundary-artifact free wavefront reconstruction from its derivatives
P. Bon, S. Monneret, B. Wattellier
Applied Optics 51 (23), 5698-5704 (2012)
Optical detection and measurement of living cell morphometric features with single-shot quantitative phase microscopy
P. Bon, J. Savatier, M. Merlin, B. Wattellier, S. Monneret
Journal of Biomedical Optics 17 (7), 076004 (2012)
Modeling quantitative phase image formation under tilted illuminations
P. Bon, B. Wattellier, S. Monneret
Optics Letters 37 (10), 1718-1720 (2012)
Selected in Virtual Journal for Biomedical Optics 7 (7), 2012
Tomographic diffractive microscopy with a wavefront sensor
Y. Ruan, P. Bon, E. Mudry, G. Maire, P. Chaumet, H. Giovannini, K. Belkebir, A. Talneau, B. Wattellier, S. Monneret, A. Sentenac
Optics Letters 37 (10), 1631-1633 (2012)
Selected in Virtual Journal for Biomedical Optics 7 (7), 2012
Quantitative absorption spectroscopy of nano-objects
P. Berto, E. Bermúdes Ureña, P. Bon, R. Quidant, H. Rigneault, G. Baffou
Physical Review B 86, 165417 (2012)
Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells
P. Bon, G. Maucort, B. Wattellier, S. Monneret
Optics Express 17 (15), 13080-13094 (2009)
Selected in Virtual Journal for Biomedical Optics 4 (9), 2009