Researching | Methods and Applications of Miniature Two-Photon Microscopes and Endomicroscopes (Invited)

Laser & Optoelectronics Progress, Volume. 62, Issue 18, 1817008(2025)

Methods and Applications of Miniature Two-Photon Microscopes and Endomicroscopes (Invited)

Kaiyan Li^1,2, Yankan Huang^1,2, Zixian Cao^1,2, Haiyan Huang^1,2, and Wenxuan Liang^1,2,3、*

Author Affiliations

¹School of Biomedical Engineering, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, Anhui , China

²Suzhou Institute for Advanced Research, University of Science and Technology of China, Suzhou 215123, Jiangsu , China

³School of Physical Sciences, University of Science and Technology of China, Hefei 230026, Anhui , China

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References(162)

[1] So P T C, Dong C Y, Masters B R et al. Two-photon excitation fluorescence microscopy[J]. Annual Review of Biomedical Engineering, 2, 399-429(2000).

[2] Zipfel W R, Williams R M, Webb W W. Nonlinear magic: multiphoton microscopy in the biosciences[J]. Nature Biotechnology, 21, 1369-1377(2003).

[3] Campagnola P J, Loew L M. Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms[J]. Nature Biotechnology, 21, 1356-1360(2003).

[4] Grienberger C, Giovannucci A, Zeiger W et al. Two-photon calcium imaging of neuronal activity[J]. Nature Reviews: Methods Primers, 2, 67(2022).

[5] Skala M C, Riching K M, Gendron-Fitzpatrick A et al. In vivo multiphoton microscopy of NADH and FAD redox states, fluorescence lifetimes, and cellular morphology in precancerous epithelia[J]. Proceedings of the National Academy of Sciences of the United States of America, 104, 19494-19499(2007).

[6] Skala M C, Riching K M, Bird D K et al. In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia[J]. Journal of Biomedical Optics, 12, 024014(2007).

[7] Georgakoudi I, Quinn K P. Optical imaging using endogenous contrast to assess metabolic state[J]. Annual Review of Biomedical Engineering, 14, 351-367(2012).

[8] Vergen J, Hecht C, Zholudeva L V et al. Metabolic imaging using two-photon excited NADH intensity and fluorescence lifetime imaging[J]. Microscopy and Microanalysis, 18, 761-770(2012).

[9] Walsh A J, Cook R S, Manning H C et al. Optical metabolic imaging identifies glycolytic levels, subtypes, and early-treatment response in breast cancer[J]. Cancer Research, 73, 6164-6174(2013).

[10] Yaseen M A, Sakadžić S, Wu W C et al. In vivo imaging of cerebral energy metabolism with two-photon fluorescence lifetime microscopy of NADH[J]. Biomedical Optics Express, 4, 307-321(2013).

[11] Hato T, Winfree S, Day R et al. Two-photon intravital fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures[J]. Journal of the American Society of Nephrology, 28, 2420-2430(2017).

[12] Kolenc O I, Quinn K P. Evaluating cell metabolism through autofluorescence imaging of NAD(P)H and FAD[J]. Antioxidants & Redox Signaling, 30, 875-889(2019).

[13] Dunn K W, Sandoval R M, Kelly K J et al. Functional studies of the kidney of living animals using multicolor two-photon microscopy[J]. American Journal of Physiology: Cell Physiology, 283, C905-C916(2002).

[14] Hall A M, Unwin R J, Parker N et al. Multiphoton imaging reveals differences in mitochondrial function between nephron segments[J]. Journal of the American Society of Nephrology, 20, 1293-1302(2009).

[15] Hall A M, Rhodes G J, Sandoval R M et al. In vivo multiphoton imaging of mitochondrial structure and function during acute kidney injury[J]. Kidney International, 83, 72-83(2013).

[16] Ranjit S, Lanzanò L, Libby A E et al. Advances in fluorescence microscopy techniques to study kidney function[J]. Nature Reviews Nephrology, 17, 128-144(2020).

[17] Provenzano P P, Eliceiri K W, Campbell J M et al. Collagen reorganization at the tumor-stromal interface facilitates local invasion[J]. BMC Medicine, 4, 38(2006).

[18] Provenzano P P, Inman D R, Eliceiri K W et al. Collagen density promotes mammary tumor initiation and progression[J]. BMC Medicine, 6, 11(2008).

[19] Akins M L, Luby-Phelps K, Mahendroo M. Second harmonic generation imaging as a potential tool for staging pregnancy and predicting preterm birth[J]. Journal of Biomedical Optics, 15, 026020(2010).

[20] James D S, Campagnola P J. Recent advancements in optical harmonic generation microscopy: applications and perspectives[J]. BME Frontiers, 2021, 3973857(2021).

[21] Bishop K W, Maitland K C, Rajadhyaksha M et al. In vivo microscopy as an adjunctive tool to guide detection, diagnosis, and treatment[J]. Journal of Biomedical Optics, 27, 040601(2022).

[22] Agrawal G P[M]. Nonlinear fiber optics(2001).

[23] Weiner A[M]. Ultrafast optics(2011).

[24] Helmchen F, Fee M S, Tank D W et al. A miniature head-mounted two-photon microscope high-resolution brain imaging in freely moving animals[J]. Neuron, 31, 903-912(2001).

[25] Rivera D R, Brown C M, Ouzounov D G et al. Compact and flexible raster scanning multiphoton endoscope capable of imaging unstained tissue[J]. Proceedings of the National Academy of Sciences of the United States of America, 108, 17598-17603(2011).

[26] Zhang Y Y, Akins M L, Murari K et al. A compact fiber-optic SHG scanning endomicroscope and its application to visualize cervical remodeling during pregnancy[J]. Proceedings of the National Academy of Sciences of the United States of America, 109, 12878-12883(2012).

[27] Oberthaler M, Höpfel R A. Special narrowing of ultrashort laser pulses by self‐phase modulation in optical fibers[J]. Applied Physics Letters, 63, 1017-1019(1993).

[28] Planas S A, Mansur N L, Cruz C H et al. Spectral narrowing in the propagation of chirped pulses in single-mode fibers[J]. Optics Letters, 18, 699-701(1993).

[29] Clark S W, Ilday F Ö, Wise F W. Fiber delivery of femtosecond pulses from a Ti: sapphire laser[J]. Optics Letters, 26, 1320(2001).

[30] Lelek M, Suran E, Louradour F et al. Coherent femtosecond pulse shaping for the optimization of a non-linear micro-endoscope[J]. Optics Express, 15, 10154-10162(2007).

[31] Lefort C, Mansuryan T, Louradour F et al. Pulse compression and fiber delivery of 45 fs Fourier transform limited pulses at 830 nm[J]. Optics Letters, 36, 292-294(2011).

[32] Liang W X, Hall G, Li X D. Spectro-temporal dispersion management of femtosecond pulses for fiber-optic two-photon endomicroscopy[J]. Optics Express, 26, 22877-22893(2018).

[33] Gibson E A, Gaudiosi D M, Kapteyn H C et al. Efficient reflection grisms for pulse compression and dispersion compensation of femtosecond pulses[J]. Optics Letters, 31, 3363-3365(2006).

[34] Li A, Guan H H, Park H C et al. Twist-free ultralight two-photon fiberscope enabling neuroimaging on freely rotating/walking mice[J]. Optica, 8, 870-879(2021).

[35] Udovich J A, Kirkpatrick N D, Kano A et al. Spectral background and transmission characteristics of fiber optic imaging bundles[J]. Applied Optics, 47, 4560-4568(2008).

[36] Chang Y C, Ye J Y, Baker J R et al. Two-photon fluorescence correlation spectroscopy through a dual-clad optical fiber[C], 1-2(2008).

[37] Ducourthial G, Leclerc P, Mansuryan T et al. Development of a real-time flexible multiphoton microendoscope for label-free imaging in a live animal[J]. Scientific Reports, 5, 18303(2015).

[38] Liang W X, Hall G, Messerschmidt B et al. Nonlinear optical endomicroscopy for label-free functional histology in vivo[J]. Light: Science & Applications, 6, e17082(2017).

[39] Knight J C, Birks T A, Russell P St J et al. All-silica single-mode optical fiber with photonic crystal cladding[J]. Optics Letters, 21, 1547-1549(1996).

[40] Birks T A, Knight J C, Russell P J, St. Endlessly single-mode photonic crystal fiber[J]. Optics Letters, 22, 961-963(1997).

[41] Fu L, Jain A, Xie H K et al. Nonlinear optical endoscopy based on a double-clad photonic crystal fiber and a MEMS mirror[J]. Optics Express, 14, 1027-1032(2006).

[42] Fu L, Jain A, Cranfield C et al. Three-dimensional nonlinear optical endoscopy[J]. Journal of Biomedical Optics, 12, 040501(2007).

[43] Fu L, Gu M. Fibre-optic nonlinear optical microscopy and endoscopy[J]. Journal of Microscopy, 226, 195-206(2007).

[44] Cregan R F, Mangan B J, Knight J C et al. Single-mode photonic band gap guidance of light in air[J]. Science, 285, 1537-1539(1999).

[45] Smith C M, Venkataraman N, Gallagher M T et al. Low-loss hollow-core silica/air photonic bandgap fibre[J]. Nature, 424, 657-659(2003).

[46] Amezcua-Correa R, Broderick N G R, Petrovich M N et al. Optimizing the usable bandwidth and loss through core design in realistic hollow-core photonic bandgap fibers[J]. Optics Express, 14, 7974-7985(2006).

[47] Poletti F, Petrovich M N, Richardson D J. Hollow-core photonic bandgap fibers: technology and applications[J]. Nanophotonics, 2, 315-340(2013).

[48] Zong W J, Wu R L, Li M L et al. Fast high-resolution miniature two-photon microscopy for brain imaging in freely behaving mice[J]. Nature Methods, 14, 713-719(2017).

[49] Wu Y C, Leng Y X, Xi J F et al. Scanning all-fiber-optic endomicroscopy system for 3D nonlinear optical imaging of biological tissues[J]. Optics Express, 17, 7907-7915(2009).

[50] Couny F, Benabid F, Roberts P J et al. Generation and photonic guidance of multi-octave optical-frequency combs[J]. Science, 318, 1118-1121(2007).

[51] Wei C L, Joseph Weiblen R, Menyuk C R et al. Negative curvature fibers[J]. Advances in Optics and Photonics, 9, 504-561(2017).

[52] Lombardini A, Mytskaniuk V, Sivankutty S et al. High-resolution multimodal flexible coherent Raman endoscope[J]. Light: Science & Applications, 7, 10(2018).

[53] Kudlinski A, Cassez A, Vanvincq O et al. Double clad tubular anti-resonant hollow core fiber for nonlinear microendoscopy[J]. Optics Express, 28, 15062-15070(2020).

[54] Septier D, Mytskaniuk V, Habert R et al. Label-free highly multimodal nonlinear endoscope[J]. Optics Express, 30, 25020-25033(2022).

[55] Helmchen F, Denk W. Deep tissue two-photon microscopy[J]. Nature Methods, 2, 932-940(2005).

[56] Sofroniew N J, Flickinger D, King J et al. A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging[J]. eLife, 5, e14472(2016).

[57] Matz G, Messerschmidt B, Gross H. Design and evaluation of new color-corrected rigid endomicroscopic high NA GRIN-objectives with a sub-micron resolution and large field of view[J]. Optics Express, 24, 10987-11001(2016).

[58] Jung J C, Mehta A D, Aksay E et al. In vivo mammalian brain imaging using one- and two-photon fluorescence microendoscopy[J]. Journal of Neurophysiology, 92, 3121-3133(2004).

[59] Myaing M T, MacDonald D J, Li X D. Fiber-optic scanning two-photon fluorescence endoscope[J]. Optics Letters, 31, 1076-1078(2006).

[60] Hoy C L, Durr N J, Chen P Y et al. Miniaturized probe for femtosecond laser microsurgery and two-photon imaging[J]. Optics Express, 16, 9996-10005(2008).

[61] Engelbrecht C J, Johnston R S, Seibel E J et al. Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo[J]. Optics Express, 16, 5556-5564(2008).

[62] Flusberg B A, Jung J C, Cocker E D et al. In vivo brain imaging using a portable 3.9 gram two-photon fluorescence microendoscope[J]. Optics Letters, 30, 2272-2274(2005).

[63] Piyawattanametha W, Cocker E D, Burns L D et al. In vivo brain imaging using a portable 2.9 g two-photon microscope based on a microelectromechanical systems scanning mirror[J]. Optics Letters, 34, 2309-2311(2009).

[64] Wu Y C, Xi J F, Cobb M J et al. Scanning fiber-optic nonlinear endomicroscopy with miniature aspherical compound lens and multimode fiber collector[J]. Optics Letters, 34, 953-955(2009).

[65] Zhou Z C, Gordon-Fennell A, Piantadosi S C et al. Deep-brain optical recording of neural dynamics during behavior[J]. Neuron, 111, 3716-3738(2023).

[66] Li A, Hall G, Chen D F et al. A biopsy-needle compatible varifocal multiphoton rigid probe for depth-resolved optical biopsy[J]. Journal of Biophotonics, 12, e201800229(2019).

[67] Zhao Y B, Nakamura H, Gordon R J. Development of a versatile two-photon endoscope for biological imaging[J]. Biomedical Optics Express, 1, 1159-1172(2010).

[68] Li G M, Duan X Y, Lee M et al. Ultra-compact microsystems-based confocal endomicroscope[J]. IEEE Transactions on Medical Imaging, 39, 2406-2414(2020).

[69] Wu D L, Feng L S, Wang A M. Optical design of two-photon endoscopy objective with high collection efficiency[J]. Laser & Optoelectronics Progress, 55, 071801(2018).

[70] Zhao C Z, Zhu Y F, Zhang D et al. Millimeter field-of-view miniature two-photon microscopy for brain imaging in freely moving mice[J]. Optics Express, 31, 32925-32934(2023).

[71] Barretto R P J, Messerschmidt B, Schnitzer M J. In vivo fluorescence imaging with high-resolution microlenses[J]. Nature Methods, 6, 511-512(2009).

[72] Le Harzic R, Weinigel M, Riemann I et al. Nonlinear optical endoscope based on a compact two axes piezo scanner and a miniature objective lens[J]. Optics Express, 16, 20588-20596(2008).

[73] Le Harzic R, Riemann I, Weinigel M et al. Rigid and high-numerical-aperture two-photon fluorescence endoscope[J]. Applied Optics, 48, 3396-3400(2009).

[74] Brown C M, Rivera D R, Ouzounov D G et al. In vivo imaging of unstained tissues using a compact and flexible multiphoton microendoscope[J]. Journal of Biomedical Optics, 17, 040505(2012).

[75] Liang W X, Murari K, Zhang Y Y et al. Increased illumination uniformity and reduced photodamage offered by the Lissajous scanning in fiber-optic two-photon endomicroscopy[J]. Journal of Biomedical Optics, 17, 021108(2012).

[76] Wang Y, Li Z, Liang X B et al. Four-plate piezoelectric actuator driving a large-diameter special optical fiber for nonlinear optical microendoscopy[J]. Optics Express, 24, 19949-19960(2016).

[77] Lukic A, Dochow S, Bae H et al. Endoscopic fiber probe for nonlinear spectroscopic imaging[J]. Optica, 4, 496-501(2017).

[78] Pshenay-Severin E, Bae H, Reichwald K et al. Multimodal nonlinear endomicroscopic imaging probe using a double-core double-clad fiber and focus-combining micro-optical concept[J]. Light: Science & Applications, 10, 207(2021).

[79] Li Z Y, Lin P, Huang Y W et al. Meta-optics achieves RGB-achromatic focusing for virtual reality[J]. Science Advances, 7, eabe4458(2021).

[80] Luo Y, Chu C H, Vyas S et al. Varifocal metalens for optical sectioning fluorescence microscopy[J]. Nano Letters, 21, 5133-5142(2021).

[81] Pan M Y, Fu Y F, Zheng M J et al. Dielectric metalens for miniaturized imaging systems: progress and challenges[J]. Light: Science & Applications, 11, 195(2022).

[82] Arbabi E, Li J Q, Hutchins R J et al. Two-photon microscopy with a double-wavelength metasurface objective lens[J]. Nano Letters, 18, 4943-4948(2018).

[83] Sun D Q, Yang Y J, Liu S J et al. Excitation and emission dual-wavelength confocal metalens designed directly in the biological tissue environment for two-photon micro-endoscopy[J]. Biomedical Optics Express, 11, 4408-4418(2020).

[84] Chen X H, Jin L, Chen C et al. Single-wavelength focusing metalens for multichannel two-photon microscopy[J]. IEEE Photonics Technology Letters, 35, 1407-1410(2023).

[85] Wang C H, Chen Q M, Liu H L et al. Miniature two-photon microscopic imaging using dielectric metalens[J]. Nano Letters, 23, 8256-8263(2023).

[86] Mertz J[M]. Introduction to optical microscopy(2009).

[87] Sawinski J, Denk W. Miniature random-access fiber scanner for in vivo multiphoton imaging[J]. Journal of Applied Physics, 102, 034701(2007).

[88] Sawinski J, Wallace D J, Greenberg D S et al. Visually evoked activity in cortical cells imaged in freely moving animals[J]. Proceedings of the National Academy of Sciences of the United States of America, 106, 19557-19562(2009).

[89] Zong W J, Wu R L, Chen S Y et al. Miniature two-photon microscopy for enlarged field-of-view, multi-plane and long-term brain imaging[J]. Nature Methods, 18, 46-49(2021).

[90] Camli B, Andrus L, Roy A et al. Two photon imaging probe with highly efficient autofluorescence collection at high scattering and deep imaging conditions[J]. Biomedical Optics Express, 15, 3163-3182(2024).

[91] Saar B G, Freudiger C W, Reichman J et al. Video-rate molecular imaging in vivo with stimulated Raman scattering[J]. Science, 330, 1368-1370(2010).

[92] Saar B G, Johnston R S, Freudiger C W et al. Coherent Raman scanning fiber endoscopy[J]. Optics Letters, 36, 2396-2398(2011).

[93] Utzinger U, Richards-Kortum R R. Fiber optic probes for biomedical optical spectroscopy[J]. Journal of Biomedical Optics, 8, 121-147(2003).

[94] Park H C, Seo Y H, Jeong K H. Lissajous fiber scanning for forward viewing optical endomicroscopy using asymmetric stiffness modulation[J]. Optics Express, 22, 5818-5825(2014).

[95] Park H C, Seo Y H, Hwang K et al. Micromachined tethered silicon oscillator for an endomicroscopic Lissajous fiber scanner[J]. Optics Letters, 39, 6675-6678(2014).

[96] Kim D Y, Hwang K, Ahn J et al. Lissajous scanning two-photon endomicroscope for in vivo tissue imaging[J]. Scientific Reports, 9, 3560(2019).

[97] Hwang K, Seo Y H, Kim D Y et al. Handheld endomicroscope using a fiber-optic harmonograph enables real-time and in vivo confocal imaging of living cell morphology and capillary perfusion[J]. Microsystems & Nanoengineering, 6, 72(2020).

[98] Zhang N, Tsai T H, Ahsen O O et al. Compact piezoelectric transducer fiber scanning probe for optical coherence tomography[J]. Optics Letters, 39, 186-188(2014).

[99] Zong W J, Obenhaus H A, Skytøen E R et al. Large-scale two-photon calcium imaging in freely moving mice[J]. Cell, 185, 1240-1256(2022).

[100] Holmström S T S, Baran U, Urey H. MEMS laser scanners: a review[J]. Journal of Microelectromechanical Systems, 23, 259-275(2014).

[101] Tang S, Jung W, McCormick D et al. Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning[J]. Journal of Biomedical Optics, 14, 034005(2009).

[102] Ghosh K K, Burns L D, Cocker E D et al. Miniaturized integration of a fluorescence microscope[J]. Nature Methods, 8, 871-878(2011).

[103] Bumstead J R, Park J J, Rosen I A et al. Designing a large field-of-view two-photon microscope using optical invariant analysis[J]. Neurophotonics, 5, 025001(2018).

[104] Lohmann A W, Dorsch R G, Mendlovic D et al. Space-bandwidth product of optical signals and systems[J]. Journal of the Optical Society of America A, 13, 470-473(1996).

[105] Klioutchnikov A, Wallace D J, Frosz M H et al. Three-photon head-mounted microscope for imaging deep cortical layers in freely moving rats[J]. Nature Methods, 17, 509-513(2020).

[106] Kinsler L E, Frey A R, Coppens A B et al[M]. Fundamentals of acoustics(2000).

[107] Liang W X, Park H C, Li K Y et al. Throughput-speed product augmentation for scanning fiber-optic two-photon endomicroscopy[J]. IEEE Transactions on Medical Imaging, 39, 3779-3787(2020).

[108] Wang C H, Liu H L, Cui H D et al. Two-photon endomicroscopy with microsphere-spliced double-cladding antiresonant fiber for resolution enhancement[J]. Optics Express, 30, 26090-26101(2022).

[109] McLeod E, Arnold C B. Mechanics and refractive power optimization of tunable acoustic gradient lenses[J]. Journal of Applied Physics, 102, 033104(2007).

[110] Chen L H, Ghilardi M, Busfield J J C et al. Electrically tunable lenses: a review[J]. Frontiers in Robotics and AI, 8, 678046(2021).

[111] Wu Y C, Zhang Y Y, Xi J F et al. Fiber-optic nonlinear endomicroscopy with focus scanning by using shape memory alloy actuation[J]. Journal of Biomedical Optics, 15, 060506(2010).

[112] Li A, Liang W X, Guan H H et al. Focus scanning with feedback-control for fiber-optic nonlinear endomicroscopy[J]. Biomedical Optics Express, 8, 2519-2527(2017).

[113] Ozbay B N, Futia G L, Ma M et al. Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning[J]. Scientific Reports, 8, 8108(2018).

[114] Elson D, Requejo-Isidro J, Munro I et al. Time-domain fluorescence lifetime imaging applied to biological tissue[J]. Photochemical & Photobiological Sciences, 3, 795-801(2004).

[115] Datta R, Heaster T M, Sharick J T et al. Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications[J]. Journal of Biomedical Optics, 25, 1-43(2020).

[116] Lakowicz J R, Szmacinski H, Nowaczyk K et al. Fluorescence lifetime imaging of free and protein-bound NADH[J]. Proceedings of the National Academy of Sciences of the United States of America, 89, 1271-1275(1992).

[117] Vishwasrao H D, Heikal A A, Kasischke K A et al. Conformational dependence of intracellular NADH on metabolic state revealed by associated fluorescence anisotropy[J]. Journal of Biological Chemistry, 280, 25119-25126(2005).

[118] Bird D K, Yan L, Vrotsos K M et al. Metabolic mapping of MCF10A human breast cells via multiphoton fluorescence lifetime imaging of the coenzyme NADH[J]. Cancer Research, 65, 8766-8773(2005).

[119] Chia T H, Williamson A, Spencer D D et al. Multiphoton fluorescence lifetime imaging of intrinsic fluorescence in human and rat brain tissue reveals spatially distinct NADH binding[J]. Optics Express, 16, 4237-4249(2008).

[120] Liu Z Y, Pouli D, Alonzo C A et al. Mapping metabolic changes by noninvasive, multiparametric, high-resolution imaging using endogenous contrast[J]. Science Advances, 4, eaap9302(2018).

[121] Becker W, Bergmann A, Hink M A et al. Fluorescence lifetime imaging by time-correlated single-photon counting[J]. Microscopy Research and Technique, 63, 58-66(2004).

[122] Hage C H, Leclerc P, Fabert M et al. A readily usable two-photon fluorescence lifetime microendoscope[J]. Journal of Biophotonics, 12, e201800276(2019).

[123] Liang W X, Chen D F, Guan H H et al. Label-free metabolic imaging in vivo by two-photon fluorescence lifetime endomicroscopy[J]. ACS Photonics, 9, 4017-4029(2022).

[124] Conklin M W, Eickhoff J C, Riching K M et al. Aligned collagen is a prognostic signature for survival in human breast carcinoma[J]. The American Journal of Pathology, 178, 1221-1232(2011).

[125] Lakowicz J R, Gryczynski I, Malak H et al. Two-color two-photon excitation of fluorescence[J]. Photochemistry and Photobiology, 64, 632-635(1996).

[126] Guan H H, Liang W X, Li A et al. Multicolor fiber-optic two-photon endomicroscopy for brain imaging[J]. Optics Letters, 46, 1093-1096(2021).

[127] Xi J F, Chen Y P, Zhang Y Y et al. Integrated multimodal endomicroscopy platform for simultaneous en face optical coherence and two-photon fluorescence imaging[J]. Optics Letters, 37, 362-364(2012).

[128] Huang S H, Heikal A A, Webb W W. Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein[J]. Biophysical Journal, 82, 2811-2825(2002).

[129] Abir A H, Weckwerth L, Wilhelm A et al. Metabolic profiling of single cells by exploiting NADH and FAD fluorescence via flow cytometry[J]. Molecular Metabolism, 87, 101981(2024).

[130] Hall G, Liang W X, Bhujwalla Z M et al. SHG fiberscopy assessment of collagen morphology and its potential for breast cancer optical histology[J]. IEEE Transactions on Bio-Medical Engineering, 71, 2414-2420(2024).

[131] Hall G, Liang W X, Li X D. Fitting-free algorithm for efficient quantification of collagen fiber alignment in SHG imaging applications[J]. Biomedical Optics Express, 8, 4609-4620(2017).

[132] Akins M L, Luby-Phelps K, Bank R A et al. Cervical softening during pregnancy: regulated changes in collagen cross-linking and composition of matricellular proteins in the mouse[J]. Biology of Reproduction, 84, 1053-1062(2011).

[133] Liang W X, Liu Y H, Guan H H et al. Cervical collagen network porosity assessed by SHG endomicroscopy distinguishes preterm and normal pregnancy-a pilot study[J]. IEEE Transactions on Bio-Medical Engineering, 72, 777-785(2025).

[134] Maioli V, Boniface A, Mahou P et al. Fast in vivo multiphoton light-sheet microscopy with optimal pulse frequency[J]. Biomedical Optics Express, 11, 6012-6026(2020).

[135] Podgorski K, Ranganathan G. Brain heating induced by near-infrared lasers during multiphoton microscopy[J]. Journal of Neurophysiology, 116, 1012-1023(2016).

[136] Patterson G H, Piston D W. Photobleaching in two-photon excitation microscopy[J]. Biophysical Journal, 78, 2159-2162(2000).

[137] Hopt A, Neher E. Highly nonlinear photodamage in two-photon fluorescence microscopy[J]. Biophysical Journal, 80, 2029-2036(2001).

[138] Ji N, Magee J C, Betzig E. High-speed, low-photodamage nonlinear imaging using passive pulse splitters[J]. Nature Methods, 5, 197-202(2008).

[139] Čižmár T, Dholakia K. Exploiting multimode waveguides for pure fibre-based imaging[J]. Nature Communications, 3, 1027(2012).

[140] Papadopoulos I N, Farahi S, Moser C et al. High-resolution, lensless endoscope based on digital scanning through a multimode optical fiber[J]. Biomedical Optics Express, 4, 260-270(2013).

[141] Plöschner M, Tyc T, Čižmár T. Seeing through chaos in multimode fibres[J]. Nature Photonics, 9, 529-535(2015).

[142] Sivankutty S, Andresen E R, Cossart R et al. Ultra-thin rigid endoscope: two-photon imaging through a graded-index multi-mode fiber[J]. Optics Express, 24, 825-841(2016).

[143] Farahi S, Ziegler D, Papadopoulos I N et al. Dynamic bending compensation while focusing through a multimode fiber[J]. Optics Express, 21, 22504-22514(2013).

[144] Boonzajer Flaes D E, Stopka J, Turtaev S et al. Robustness of light-transport processes to bending deformations in graded-index multimode waveguides[J]. Physical Review Letters, 120, 233901(2018).

[145] Zhao J, Hu X W, Gausmann S et al. Learning-based image transport through disordered optical fibers with transverse Anderson localization[J]. Frontiers in Physics, 9, 710351(2021).

[146] Karbasi S, Mirr C R, Yarandi P G et al. Observation of transverse Anderson localization in an optical fiber[J]. Optics Letters, 37, 2304-2306(2012).

[147] Karbasi S, Frazier R J, Koch K W et al. Image transport through a disordered optical fibre mediated by transverse Anderson localization[J]. Nature Communications, 5, 3362(2014).

[148] Patel K B, Liang W X, Casper M J et al. High-speed light-sheet microscopy for the in situ acquisition of volumetric histological images of living tissue[J]. Nature Biomedical Engineering, 6, 569-583(2022).

[149] Dunsby C. Optically sectioned imaging by oblique plane microscopy[J]. Optics Express, 16, 20306-20316(2008).

[150] Bouchard M B, Voleti V, Mendes C S et al. Swept confocally-aligned planar excitation (SCAPE) microscopy for high-speed volumetric imaging of behaving organisms[J]. Nature Photonics, 9, 113-119(2015).

[151] Power R M, Huisken J. A guide to light-sheet fluorescence microscopy for multiscale imaging[J]. Nature Methods, 14, 360-373(2017).

[152] Voleti V, Patel K B, Li W Z et al. Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0[J]. Nature Methods, 16, 1054-1062(2019).

[153] Stelzer E H K, Strobl F, Chang B J et al. Light sheet fluorescence microscopy[J]. Nature Reviews Methods Primers, 1, 73(2021).

[154] Abrahamsson S, Usawa S, Gustafsson M. A new approach to extended focus for high-speed high-resolution biological microscopy[J]. Proceedings of SPIE, 6090, 60900N(2006).

[155] Botcherby E J, Juškaitis R, Wilson T. Scanning two photon fluorescence microscopy with extended depth of field[J]. Optics Communications, 268, 253-260(2006).

[156] Thériault G, de Koninck Y, McCarthy N. Extended depth of field microscopy for rapid volumetric two-photon imaging[J]. Optics Express, 21, 10095-10104(2013).

[157] Chen B Y, Chakraborty T, Daetwyler S et al. Extended depth of focus multiphoton microscopy via incoherent pulse splitting[J]. Biomedical Optics Express, 11, 3830-3842(2020).

[158] Beaulieu D R, Davison I G, Kılıç K et al. Simultaneous multiplane imaging with reverberation two-photon microscopy[J]. Nature Methods, 17, 283-286(2020).

[159] Demas J, Manley J, Tejera F et al. High-speed, cortex-wide volumetric recording of neuroactivity at cellular resolution using light beads microscopy[J]. Nature Methods, 18, 1103-1111(2021).

[160] Belthangady C, Royer L A. Applications, promises, and pitfalls of deep learning for fluorescence image reconstruction[J]. Nature Methods, 16, 1215-1225(2019).

[161] Guan H H, Li D W, Park H C et al. Deep-learning two-photon fiberscopy for video-rate brain imaging in freely-behaving mice[J]. Nature Communications, 13, 1534(2022).

[162] Li K Y, Yang J Y, Liang W X et al. O-PRESS: Boosting OCT axial resolution with Prior guidance, Recurrence, and Equivariant Self-Supervision[J]. Medical Image Analysis, 99, 103319(2025).

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Kaiyan Li, Yankan Huang, Zixian Cao, Haiyan Huang, Wenxuan Liang. Methods and Applications of Miniature Two-Photon Microscopes and Endomicroscopes (Invited)[J]. Laser & Optoelectronics Progress, 2025, 62(18): 1817008

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Paper Information

Category: Medical Optics and Biotechnology

Received: Jul. 24, 2025

Accepted: Aug. 19, 2025

Published Online: Sep. 15, 2025

The Author Email: Wenxuan Liang (liangwenxuan@ustc.edu.cn)

DOI:10.3788/LOP251757

CSTR:32186.14.LOP251757

Topics

laser devices and laser physics

Lasers and Laser Optics

laser manufacturing

Instrumentation, Measurement and Metrology