Second harmonic generation in skin and cornea

Second harmonic generation in skin and cornea


Marie-Claire Schanne-Klein / DR CNRS
Gaël Latour / Paris-Saclay University
Hester Colboc / M.D., AP-HP, SU
Giulia Galante / PhD student
Poncia Nyembo / PhD student

Internship proposals / Propositions de stage :

Polarization-resolved SHG structural imaging of collagen remodeling

Numerical simulations of pSHG microscopy in cornea

ANR CorMecha : Multiscale mapping and biomechanics of healthy and pathological Human corneas


Second Harmonic Generation (SHG) microscopy is an efficient technique to probe the 3D architecture of collagen, which is the main structural protein in mammals and determines the architecture of many tissues (skin, cornea, artery, bone…).  Our main recent results are the following:

- We deciphered the physical origin of SHG signal from collagen at the molecular and fibrillar scales and we measured quantitatively this response.
- We implemented polarization-resolved SHG microscopy and showed that this technique probes the fibrils orientation and the orientational disorder at sub-µm scale within tissues.
- We assessed the progression of fibrotic pathologies in lung and renal tissues and characterized the 3D organization of collagen-based tissue substitutes.
- We characterized endogenous multiphoton signals in Human corneas, we demonstrated in vivo structural imaging of rat corneas and we mapped the lamellae structure of Human corneas using polarization-resolved SHG microscopy.
- We combined SHG imaging and mechanical assays to decipher the relationships between macroscopic tissue response and microscopic structure in skin and cornea.
- We implemented circular-dichroism SHG microscopy and demonstrated that it probes the polarity distribution of collagen fibrils.
- We correlated multiphoton microscopy to Infra-Red nanoscopy to characterize the denaturation of collagen to gelatin (see also application to parchments degradation).

Selected recent publications

[All publications on HAL]

Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy, C. Raoux, A. Chessel, P. Mahou, G. Latour, and M.-C. Schanne-Klein, Light: Science and Applications 12, 190 (2023). 10.1038/s41377-023-01224-0

Kidney stone classification using multimodal multiphoton microscopy, M. Gleeson, J. Morizet, P. Mahou, M. Daudon, D. Bazin, C. Stringari, M.-C. Schanne-Klein, and E. Beaurepaire, ACS Photonics 10, 3594 (2023). 10.1021/acsphotonics.3c00651

Structural imaging of keratoconic corneas using polarization-resolved SHG microscopy , C. Raoux, M. Schmeltz, M. Bied, M. Alnawaiseh, U. Hansen, G. Latour, and M.-C. Schanne-Klein, Biomed. Opt. Express 12, 4163-4178 (2021) - 10.1364/BOE.426145

Electric field measurements in plasmas: how focusing strongly distorts the E-FISH signal , T.L. Chng, S. Starikovskaia, M.-C. Schanne-Klein, Plasma Sources Sci. Technol. 29, 125002 (2020) - 10.1088/1361-6595/abbf93

Native collagen: electrospinning of pure, cross-linker free, self-supported membrane, D. Dems, J. Rodrigues da Silva, C. Hélary, F. Wien, M. Marchand, N. Debons, L. Muller, Y. Chen, M.-C. Schanne-Klein, C. Laberty-Robert, N. Krins, and C. Aimé, ACS Applied Bio Materials 3, 2948-2957 (2020) - 10.1021/acsabm.0c00006

Dichroism-SHG microscopy probes the polarity distribution of collagen fibrils, M. Schmeltz, C. Teulon, M. Pinsard, U. Hansen, M. Alnawaiseh, D. Ghoubay, V. Borderie, G. Mosser, C. Aimé, G. Latour, and M.-C. Schanne-Klein, Optica 7, 1469-1476 (2020) - 10.1364/OPTICA.399246

Monitoring dynamic collagen reorganization during skin stretching with fast polarization-resolved SHG imaging, G. Ducourthial, J.-S. Affagard, M. Schmeltz, X. Solinas, M. Lopez-Poncelas, C. Bonod-Bidaud, R. Rubio-Amador, F. Ruggiero, J.-M. Allain, E. Beaurepaire, M.-C. Schanne-Klein, J. Biophot. 12, e201800336 (2019) - 10.1002/jbio.201800336

High-speed polarization-resolved third harmonic microscopy, J. Morizet, G. Ducourthial, W. Supatto, A. Boutillon, R. Legouis, M.-C. Schanne Klein, C. Stringari, E. Beaurepaire, Optica 6, 385-388 (2019) - 10.1364/OPTICA.6.000385

Highlights / review article

[French] Cartographier la structure de la cornée humaine, Actualités scientifiques de l'Ecole Polytechnique et de l’INP CNRS, 2023
[French] Voir le collagène en 3D, Actualités scientifiques de l'Ecole Polytechnique et de l’INP CNRS, 2020
[English] Combination of Traction Assays and Multiphoton Imaging to Quantify Skin Biomechanics, Schanne-Klein M.-C. et al, In "Collagen: Methods and Protocols", I. Sagi and N. A. Afratis Eds (Springer Science, 2019)
[English] Multiscale Characterization of Skin Mechanics through in Situ Imaging, 5. J.-M. Allain, B. Lynch, M.-C. Schanne-Klein, In "Skin Biophysics - From Experimental Characterisation to Advanced Modelling", G. Limbert, ed (Springer Nature, 2019)
[English] SHG imaging and quantization of fibrosis, Schanne-Klein M.-C., In "Second Harmonic Generation Imaging", F. S. Pavone and P. J. Campagnola Editors, p.349-371 (Taylor and Francis, 2013)

Main ongoing collaborations

- C. Aimé, ENS Paris (CNRS, SU)
- J.-M. Allain, LMS (CNRS, Ecole Polytechnique - Palaiseau)
- A. Dazzi, A. Deniset-Besseau, ICP (CNRS, Université Paris-Sud - Orsay)
- V. Borderie, K. Grieve, D. Ghoubay, Hôpital des 15-20, Paris
- U. Hansen, University-Hospital Muenster, Germany
- F. Légaré, INRS, Montréal, Canada

Alumni/PhD students

- Thierry Boulesteix : Microscopie non linéaire: chiralité et génération de second harmonique (07/2004) - Professeur en classes préparatoires
- Ana-Maria Pena : Génération de seconde harmonique par le collagène et application à l'étude de fibroses par microscopie multiphoton (09/2006) - Research Engineer at L'Oreal-Advanced research
- M. Strupler : Imagerie du collagène par microscopie multiphotonique. Application aux fibroses rénales (11/2008) - Research Associate at Polytechnique Montreal
- Ivan Gusachenko : Polarization-resolved Second Harmonic Microscopy in collagenous tissues (10/2012) - Research engineer at Cailabs, France
- Stéphane Bancelin : Imagerie quantitative du collagène par imagerie de seconde harmonique (12/2013) - CNRS permanent position at Bordeaux, France
- Claire Teulon: Imagerie quantitative du collagène par microscopie par génération de second harmonique résolue en polarisation (10/2016) - Professeur de lycée
- Margaux Schmeltz: Microscopie par génération de second harmonique (SHG) résolue en polarisations linéaire et circulaire pour caractériser l’organisation 3D du collagène (10/2019)
- Clothilde Raoux: Caractérisation de la structure de la cornée humaine par microscopie de second harmonique résolue en polarisation (12/2021)


- Ariane Deniset-Besseau, 2008-2009 - Associate professor at Paris-Sud University
- Yannick Goulam Houssen, 2009 - Research Engineer at Paris 5 University
- Gaël Latour, 2010-2012 - Associate professor at Paris-Sud University
- Sotiris Psilodimitrakopoulos, 2013-2014 - Foundation for Research and Technology, Irákleion, Greece
- Guillaume Ducourthial, 2015-2016 - research engineer at Horiba-Jobin-Yvon, France