Brain-wide multicolor 2P microscopy

Brain-wide multicolor 2P microscopy

The study of brain tissue development requires imaging approaches that ideally provide micrometer resolution, access to mm3 volumes, and multi-contrast capability. We develop methods for imaging multiple structural and biological parameters in complex tissues.

Multicolor two-photon imaging: we introduced a novel approach for simultaneous two-photon imaging of several fluorescent proteins (e.g. CFP, YFP, RFP) in tissues. This method, which is based on nonlinear wavelength mixing, is particularly appropriate for 'brainbow' tissue imaging, which we develop in collaboration with J. Livet's group at Institut de la Vision. By combining this method with automated serial sectioning, we extended it to brain-wide multicolor imaging with micrometric resolution (ChroMS, chromatic multiphoton serial microscopy) in brainbow mice. 

Multimodal imaging:
we optimize multimodal tissue imaging approaches simultaneously providing complementary information based on fluorescence (2P, 3P), harmonic generation (SHG, THG), wave-mixing (TSFG, FWM, CARS), and/or fluorescence lifetime (FLIM) signals.

Related publications

Chromatically corrected multicolor multiphoton microscopy
H. Blanc, G. Kaddour, N.B. David, W. Supatto, J. Livet, E. Beaurepaire, P. Mahou
ACS Photonics (2023)
Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy
L. Abdeladim, K.S. Matho, S. Clavreul, P. Mahou, J.-M. Sintes, X. Solinas, I. Arganda-Carreras, S.G. Turney, J.W. Lichtman, A. Chessel, A.-P. Bemelmans, K. Loulier, W. Supatto, J. Livet, E. Beaurepaire
Nature Communications 10, 1662 (2019)
Cortical astrocytes develop in a plastic manner at both clonal and cellular levels
S. Clavreul, L. Abdeladim, E. Hernández-Garzón, D.Niculescu, J. Durand, S.-H. Ieng, R. Barry, G. Bonvento, E. Beaurepaire, J. Livet, K. Loulier
Nature Communications 10, 4884 (2019)
Multicolor two-photon imaging of endogenous fluorophores in living tissues by wavelength mixing C. Stringari, L. Abdeladim, G. Malkinson, P. Mahou, X. Solinas, I. Lamarre, S. Brizion, J.-B. Galey, W. Supatto, R. Legouis, A.-M. Pena, E. Beaurepaire
Scientific Reports 7, 3792 (2017).
Metrology of multiphoton microscopes using second harmonic generation nanoprobes
P. Mahou, G. Malkinson, E. Chaudan, T. Gacoin, E. Beaurepaire, W. Supatto
Small 13, 1701442 (2017).
Life-long neurogenic activity of individual neural stem cells and continuous growth establish an outside-in architecture in the teleost pallium
G. Furlan, V. Cuccioli, N. Vuillemin, L. Dirian, A. Janue Muntasell, M. Coolen, N. Dray, S. Bedu, C. Houart, E. Beaurepaire, I. Foucher, L. Bally-Cuif
Current Biology 27, 3288-3301 (2017).
Large-scale live imaging of adult neural stem cells in their endogenous niche
N. Dray, S. Bedu, N. Vuillemin, A. Alunni, M. Coolen, M. Krecsmarik, W. Supatto, E. Beaurepaire, and L. Bally-Cuif
Development 142: 3592-3600 (2015).
Multicolor two-photon light-sheet microscopy
P. Mahou, J. vermot, E. Beaurepaire, W. Supatto
Nature Methods 11(6), 600-601 (2014).
Multiplex cell and lineage tracking with combinatorial labels
K. Loulier, R. Barry, P. Mahou, Y. Le Franc, W. Supatto, K.S. Matho, S. Ieng, S. Fouquet, E. Dupin, R. Benosman, A. Chédotal, E. Beaurepaire, X. Morin, J. Livet
Neuron 81(3), 505-520 (2014).
Multicolor two-photon tissue imaging by wavelength mixing
P. Mahou, M. Zimmerley, K. Loulier, K. Matho, G. Labroille, X. Morin, W. Supatto, J. Livet, D. Débarre & E. Beaurepaire
Nature Methods 9(8) 815-818 (2012).
Combined third-harmonic generation and four-wave mixing microscopy of tissues and embryos
P. Mahou, N. Olivier, G. Labroille, L. Duloquin, J-M. Sintes, N. Peyriéras, R. Legouis, D. Débarre, and E. Beaurepaire
Biomed. Opt. Express 2(10), 2837-2849 (2011).
Cell lineage reconstruction of early zebrafish embryos using label-free nonlinear microscopy
N. Olivier, M. Luengo-Oroz, L. Duloquin, E. Faure, T. Savy, I. Veilleux, X. Solinas, D. Débarre, P. Bourgine, A. Santos, N. Peyriéras & E. Beaurepaire
Science 339(5994), 967-71 (2010).