Infrared multiphoton microscopy: subcellular-resolved deep tissue imaging.

Multiphoton microscopy (MPM) is the method of choice for investigating cells and cellular functions in deep tissue sections and organs. Here we present the setup and applications of infrared-(IR-)MPM using excitation wavelengths above 1080 nm. IR-MPM enables the use of red fluorophores and fluorescent proteins, doubles imaging depth, improves second harmonic generation of tissue structures, and strongly reduces phototoxicity and photobleaching, compared with conventional MPM. Furthermore, it still provides subcellular resolution at depths of several hundred micrometers and thus will enhance long-term live cell and deep tissue microscopy.

[1]  J. Schuman,et al.  Optical coherence tomography. , 2000, Science.

[2]  Stephanie Alexander,et al.  Dynamic imaging of cancer growth and invasion: a modified skin-fold chamber model , 2008, Histochemistry and Cell Biology.

[3]  J. Girkin,et al.  Practical implementation of adaptive optics in multiphoton microscopy. , 2003, Optics express.

[4]  Yaron Silberberg,et al.  Nonlinear scanning laser microscopy by third harmonic generation , 1997 .

[5]  Conor L Evans,et al.  Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[6]  R. Balaban,et al.  Skeletal muscle NAD(P)H two-photon fluorescence microscopy in vivo: topology and optical inner filters. , 2005, Biophysical journal.

[7]  Ruben M Sandoval,et al.  Functional Studies of the Kidney of Living Animals Using Multicolor 2-photon Microscopy , 2022 .

[8]  Heinrich Spiecker,et al.  The power of single and multibeam two-photon microscopy for high-resolution and high-speed deep tissue and intravital imaging. , 2007, Biophysical journal.

[9]  Wilson,et al.  3D microscopy of transparent objects using third‐harmonic generation , 1998, Journal of microscopy.

[10]  S. Henrickson,et al.  T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases , 2004, Nature.

[11]  K. Fujita [Two-photon laser scanning fluorescence microscopy]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[12]  Scott E Fraser,et al.  Multiphoton excitation spectra in biological samples. , 2003, Journal of biomedical optics.

[13]  G. Wahl,et al.  Cellular Dynamics Visualized in Live Cells in Vitro and in Vivo by Differential Dual-Color Nuclear-Cytoplasmic Fluorescent-Protein Expression , 2004, Cancer Research.

[14]  P. Török,et al.  Point-spread function reconstruction in high aperture lenses focusing ultra-short laser pulses , 2002 .

[15]  Rosa Cossart,et al.  A Parturition-Associated Nonsynaptic Coherent Activity Pattern in the Developing Hippocampus , 2007, Neuron.

[16]  W. Webb,et al.  Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Segall,et al.  Intravital imaging of cell movement in tumours , 2003, Nature Reviews Cancer.

[18]  Iris Riemann,et al.  High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution. , 2003, Journal of biomedical optics.

[19]  T. Wilson,et al.  Characterizing specimen induced aberrations for high NA adaptive optical microscopy. , 2004, Optics express.

[20]  B R Masters,et al.  Multiphoton excitation fluorescence microscopy and spectroscopy of in vivo human skin. , 1997, Biophysical journal.

[21]  B. Tromberg,et al.  Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Peter Friedl,et al.  Biological Second and Third Harmonic Generation Microscopy , 2007, Current protocols in cell biology.

[23]  W. Denk,et al.  Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing , 2006, Proceedings of the National Academy of Sciences.

[24]  K. König,et al.  Multiphoton microscopy in life sciences , 2000, Journal of microscopy.

[25]  W. Denk,et al.  Two-photon imaging to a depth of 1000 microm in living brains by use of a Ti:Al2O3 regenerative amplifier. , 2003, Optics letters.

[26]  E. Neher,et al.  Highly nonlinear photodamage in two-photon fluorescence microscopy. , 2001, Biophysical journal.

[27]  Andreas Volkmer,et al.  Molecular photobleaching kinetics of Rhodamine 6G by one- and two-photon induced confocal fluorescence microscopy. , 2005, Chemphyschem : a European journal of chemical physics and physical chemistry.

[28]  V. Dashkevich,et al.  Tunable optical parametric oscillator based on a KTP crystal, pumped by a pulsed Ti3+:Al2O3 laser , 2007 .

[29]  G. M. Hale,et al.  Optical Constants of Water in the 200-nm to 200-microm Wavelength Region. , 1973, Applied optics.

[30]  W. Denk,et al.  Deep tissue two-photon microscopy , 2005, Nature Methods.