Abstract

We demonstrate a novel miniaturized multimodal coherent anti-Stokes Raman scattering (CARS) microscope based on microelectromechanical systems (MEMS) scanning mirrors and custom miniature optics. A single Ti:sapphire femtosecond pulsed laser is used as the light source to produce the CARS, two photon excitation fluorescence (TPEF) and second harmonic generation (SHG) images using this miniaturized microscope. The high resolution and distortion-free images obtained from various samples such as a USAF target, fluorescent and polystyrene microspheres and biological tissue successfully demonstrate proof of concept, and pave the path towards future integration of parts into a handheld multimodal CARS probe for non- or minimally-invasive in vivo imaging.

© 2010 OSA

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2010

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

H. Bao, A. Boussioutas, R. Jeremy, S. Russell, and M. Gu, “Second harmonic generation imaging via nonlinear endomicroscopy,” Opt. Express 18(2), 1255–1260 (2010).
[CrossRef] [PubMed]

M. Balu, G. Liu, Z. Chen, B. J. Tromberg, and E. O. Potma, “Fiber delivered probe for efficient CARS imaging of tissues,” Opt. Express 18(3), 2380–2388 (2010).
[CrossRef] [PubMed]

2009

H. Chen, H. Wang, M. N. Slipchenko, Y. Jung, Y. Shi, J. Zhu, K. K. Buhman, and J. X. Cheng, “A multimodal platform for nonlinear optical microscopy and microspectroscopy,” Opt. Express 17(3), 1282–1290 (2009).
[CrossRef] [PubMed]

W. Piyawattanametha, E. D. Cocker, L. D. Burns, R. P. Barretto, J. C. Jung, H. Ra, O. Solgaard, and M. J. Schnitzer, “In vivo brain imaging using a portable 2.9 g two-photon microscope based on a microelectromechanical systems scanning mirror,” Opt. Lett. 34(15), 2309–2311 (2009).
[CrossRef] [PubMed]

S. Murugkar, C. L. Evans, X. S. Xie, and H. Anis, “Chemically specific imaging of cryptosporidium oocysts using coherent anti-Stokes Raman scattering (CARS) microscopy,” J. Microsc. 233(2), 244–250 (2009).
[CrossRef] [PubMed]

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

2008

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

C. L. Evans and X. S. Xie, “Coherent anti-stokes Raman scattering microscopy: chemical imaging for biology and medicine,” Annu Rev Anal Chem (Palo Alto Calif) 1(1), 883–909 (2008).
[CrossRef]

T. B. Huff, Y. Shi, Y. Fu, H. Wang, and J. X. Cheng, “Multimodal nonlinear optical microscopy and applications to central nervous system,” IEEE J. Sel. Top. Quantum Electron. 14(1), 4–9 (2008).
[CrossRef] [PubMed]

C. J. Engelbrecht, R. S. Johnston, E. J. Seibel, and F. Helmchen, “Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo,” Opt. Express 16(8), 5556–5564 (2008).
[CrossRef] [PubMed]

H. Bao, J. Allen, R. Pattie, R. Vance, and M. Gu, “Fast handheld two-photon fluorescence microendoscope with a 475 µm x 475 µm field of view for in vivo imaging,” Opt. Lett. 33(12), 1333–1335 (2008).
[CrossRef] [PubMed]

C. L. Hoy, N. J. Durr, P. Chen, W. Piyawattanametha, H. Ra, O. Solgaard, and A. Ben-Yakar, “Miniaturized probe for femtosecond laser microsurgery and two-photon imaging,” Opt. Express 16(13), 9996–10005 (2008).
[CrossRef] [PubMed]

T. M. Liu, M. C. Chan, I. H. Chen, S. H. Chia, and C. K. Sun, “Miniaturized multiphoton microscope with a 24Hz frame-rate,” Opt. Express 16(14), 10501–10506 (2008).
[CrossRef] [PubMed]

2007

H. Wang, T. B. Huff, Y. Fu, K. Y. Jia, and J. X. Cheng, “Increasing the imaging depth of coherent anti-Stokes Raman scattering microscopy with a miniature microscope objective,” Opt. Lett. 32(15), 2212–2214 (2007).
[CrossRef] [PubMed]

S. Murugkar, C. Brideau, A. Ridsdale, M. Naji, P. K. Stys, and H. Anis, “Coherent anti-Stokes Raman scattering microscopy using photonic crystal fiber with two closely lying zero dispersion wavelengths,” Opt. Express 15(21), 14028–14037 (2007).
[CrossRef] [PubMed]

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12(5), 054007 (2007).
[CrossRef] [PubMed]

L. Fu and M. Gu, “Fibre-optic nonlinear optical microscopy and endoscopy,” J. Microsc. 226(3), 195–206 (2007).
[CrossRef] [PubMed]

2006

2005

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

2003

X. L. Nan, J. X. Cheng, and X. S. Xie, “Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy,” J. Lipid Res. 44(11), 2202–2208 (2003).
[CrossRef] [PubMed]

2001

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope. high-resolution brain imaging in freely moving animals,” Neuron 31(6), 903–912 (2001).
[CrossRef] [PubMed]

Acker, H.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Ahn, Y. C.

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Allen, J.

Anis, H.

S. Murugkar, C. L. Evans, X. S. Xie, and H. Anis, “Chemically specific imaging of cryptosporidium oocysts using coherent anti-Stokes Raman scattering (CARS) microscopy,” J. Microsc. 233(2), 244–250 (2009).
[CrossRef] [PubMed]

S. Murugkar, C. Brideau, A. Ridsdale, M. Naji, P. K. Stys, and H. Anis, “Coherent anti-Stokes Raman scattering microscopy using photonic crystal fiber with two closely lying zero dispersion wavelengths,” Opt. Express 15(21), 14028–14037 (2007).
[CrossRef] [PubMed]

Balu, M.

Bao, H.

Barretto, R. P.

Ben-Yakar, A.

Berchner-Pfannschmidt, U.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Bestvater, F.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Boussioutas, A.

Bräuer, A.

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

Brideau, C.

Buckley, J.

Buhman, K. K.

Burns, L. D.

Camarillo, I. G.

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

Chan, M. C.

Chen, H.

Chen, I. H.

Chen, P.

Chen, Z.

M. Balu, G. Liu, Z. Chen, B. J. Tromberg, and E. O. Potma, “Fiber delivered probe for efficient CARS imaging of tissues,” Opt. Express 18(3), 2380–2388 (2010).
[CrossRef] [PubMed]

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Cheng, J. X.

H. Chen, H. Wang, M. N. Slipchenko, Y. Jung, Y. Shi, J. Zhu, K. K. Buhman, and J. X. Cheng, “A multimodal platform for nonlinear optical microscopy and microspectroscopy,” Opt. Express 17(3), 1282–1290 (2009).
[CrossRef] [PubMed]

T. B. Huff, Y. Shi, Y. Fu, H. Wang, and J. X. Cheng, “Multimodal nonlinear optical microscopy and applications to central nervous system,” IEEE J. Sel. Top. Quantum Electron. 14(1), 4–9 (2008).
[CrossRef] [PubMed]

H. Wang, T. B. Huff, Y. Fu, K. Y. Jia, and J. X. Cheng, “Increasing the imaging depth of coherent anti-Stokes Raman scattering microscopy with a miniature microscope objective,” Opt. Lett. 32(15), 2212–2214 (2007).
[CrossRef] [PubMed]

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12(5), 054007 (2007).
[CrossRef] [PubMed]

H. Wang, T. B. Huff, and J. X. Cheng, “Coherent anti-Stokes Raman scattering imaging with a laser source delivered by a photonic crystal fiber,” Opt. Lett. 31(10), 1417–1419 (2006).
[CrossRef] [PubMed]

X. L. Nan, J. X. Cheng, and X. S. Xie, “Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy,” J. Lipid Res. 44(11), 2202–2208 (2003).
[CrossRef] [PubMed]

Chia, S. H.

Chong, A.

Cocker, E. D.

Contag, C. H.

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

Denk, W.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope. high-resolution brain imaging in freely moving animals,” Neuron 31(6), 903–912 (2001).
[CrossRef] [PubMed]

Durr, N. J.

Engelbrecht, C. J.

Evans, C. L.

S. Murugkar, C. L. Evans, X. S. Xie, and H. Anis, “Chemically specific imaging of cryptosporidium oocysts using coherent anti-Stokes Raman scattering (CARS) microscopy,” J. Microsc. 233(2), 244–250 (2009).
[CrossRef] [PubMed]

C. L. Evans and X. S. Xie, “Coherent anti-stokes Raman scattering microscopy: chemical imaging for biology and medicine,” Annu Rev Anal Chem (Palo Alto Calif) 1(1), 883–909 (2008).
[CrossRef]

F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS Endoscopy,” Opt. Express 14(10), 4427–4432 (2006).
[CrossRef] [PubMed]

Fee, M. S.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope. high-resolution brain imaging in freely moving animals,” Neuron 31(6), 903–912 (2001).
[CrossRef] [PubMed]

Feurer, T.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Frommhagen, K.

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

Fu, L.

L. Fu and M. Gu, “Fibre-optic nonlinear optical microscopy and endoscopy,” J. Microsc. 226(3), 195–206 (2007).
[CrossRef] [PubMed]

Fu, Y.

T. B. Huff, Y. Shi, Y. Fu, H. Wang, and J. X. Cheng, “Multimodal nonlinear optical microscopy and applications to central nervous system,” IEEE J. Sel. Top. Quantum Electron. 14(1), 4–9 (2008).
[CrossRef] [PubMed]

H. Wang, T. B. Huff, Y. Fu, K. Y. Jia, and J. X. Cheng, “Increasing the imaging depth of coherent anti-Stokes Raman scattering microscopy with a miniature microscope objective,” Opt. Lett. 32(15), 2212–2214 (2007).
[CrossRef] [PubMed]

Ganikhanov, F.

Gerwig, Ch.

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

Gu, M.

Hacker, M.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Haeberle, H.

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

Heckel-Pompey, A.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Helmchen, F.

C. J. Engelbrecht, R. S. Johnston, E. J. Seibel, and F. Helmchen, “Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo,” Opt. Express 16(8), 5556–5564 (2008).
[CrossRef] [PubMed]

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope. high-resolution brain imaging in freely moving animals,” Neuron 31(6), 903–912 (2001).
[CrossRef] [PubMed]

Hoy, C. L.

Huff, T. B.

T. B. Huff, Y. Shi, Y. Fu, H. Wang, and J. X. Cheng, “Multimodal nonlinear optical microscopy and applications to central nervous system,” IEEE J. Sel. Top. Quantum Electron. 14(1), 4–9 (2008).
[CrossRef] [PubMed]

H. Wang, T. B. Huff, Y. Fu, K. Y. Jia, and J. X. Cheng, “Increasing the imaging depth of coherent anti-Stokes Raman scattering microscopy with a miniature microscope objective,” Opt. Lett. 32(15), 2212–2214 (2007).
[CrossRef] [PubMed]

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

H. Wang, T. B. Huff, and J. X. Cheng, “Coherent anti-Stokes Raman scattering imaging with a laser source delivered by a photonic crystal fiber,” Opt. Lett. 31(10), 1417–1419 (2006).
[CrossRef] [PubMed]

Jeremy, R.

Jia, K. Y.

Johnston, R. S.

Jung, J. C.

Jung, W.

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Jung, Y.

Kino, G. S.

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

Knobbe, J.

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

Lakner, H.

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

Langohr, I. M.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12(5), 054007 (2007).
[CrossRef] [PubMed]

Le, T. T.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12(5), 054007 (2007).
[CrossRef] [PubMed]

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

Lee, D.

Légaré, F.

Liu, G.

Liu, J. T. C.

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

Liu, T. M.

Locker, M. J.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12(5), 054007 (2007).
[CrossRef] [PubMed]

Loewke, N. O.

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

Maitland, K. C.

Mandella, M. J.

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

McCormick, D.

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Murugkar, S.

S. Murugkar, C. L. Evans, X. S. Xie, and H. Anis, “Chemically specific imaging of cryptosporidium oocysts using coherent anti-Stokes Raman scattering (CARS) microscopy,” J. Microsc. 233(2), 244–250 (2009).
[CrossRef] [PubMed]

S. Murugkar, C. Brideau, A. Ridsdale, M. Naji, P. K. Stys, and H. Anis, “Coherent anti-Stokes Raman scattering microscopy using photonic crystal fiber with two closely lying zero dispersion wavelengths,” Opt. Express 15(21), 14028–14037 (2007).
[CrossRef] [PubMed]

Naji, M.

Nan, X. L.

X. L. Nan, J. X. Cheng, and X. S. Xie, “Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy,” J. Lipid Res. 44(11), 2202–2208 (2003).
[CrossRef] [PubMed]

Nichols, M. B.

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

Pattie, R.

Piyawattanametha, W.

Porwol, T.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Potma, E. O.

Ra, H.

Rehrer, C. W.

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

Renninger, W.

Richards-Kortum, R.

Ridsdale, A.

Russell, S.

Schenk, H.

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

Schlebusch, D.

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

Schnitzer, M. J.

Scholles, M.

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

Schwarzenberg, M.

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

Seibel, E. J.

Shi, Y.

H. Chen, H. Wang, M. N. Slipchenko, Y. Jung, Y. Shi, J. Zhu, K. K. Buhman, and J. X. Cheng, “A multimodal platform for nonlinear optical microscopy and microspectroscopy,” Opt. Express 17(3), 1282–1290 (2009).
[CrossRef] [PubMed]

T. B. Huff, Y. Shi, Y. Fu, H. Wang, and J. X. Cheng, “Multimodal nonlinear optical microscopy and applications to central nervous system,” IEEE J. Sel. Top. Quantum Electron. 14(1), 4–9 (2008).
[CrossRef] [PubMed]

Shin, H. J.

Slipchenko, M. N.

Solgaard, O.

Spiess, E.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Stobrawa, G.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Sturek, M.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12(5), 054007 (2007).
[CrossRef] [PubMed]

Stys, P. K.

Su, J.

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Sun, C. K.

Tang, S.

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Tank, D. W.

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope. high-resolution brain imaging in freely moving animals,” Neuron 31(6), 903–912 (2001).
[CrossRef] [PubMed]

Toth, K.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Tromberg, B. J.

M. Balu, G. Liu, Z. Chen, B. J. Tromberg, and E. O. Potma, “Fiber delivered probe for efficient CARS imaging of tissues,” Opt. Express 18(3), 2380–2388 (2010).
[CrossRef] [PubMed]

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Vance, R.

Vogel, U.

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

Wang, H.

Wise, F.

Wotzlaw, C.

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

Xie, T.

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

Xie, X. S.

S. Murugkar, C. L. Evans, X. S. Xie, and H. Anis, “Chemically specific imaging of cryptosporidium oocysts using coherent anti-Stokes Raman scattering (CARS) microscopy,” J. Microsc. 233(2), 244–250 (2009).
[CrossRef] [PubMed]

C. L. Evans and X. S. Xie, “Coherent anti-stokes Raman scattering microscopy: chemical imaging for biology and medicine,” Annu Rev Anal Chem (Palo Alto Calif) 1(1), 883–909 (2008).
[CrossRef]

F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS Endoscopy,” Opt. Express 14(10), 4427–4432 (2006).
[CrossRef] [PubMed]

X. L. Nan, J. X. Cheng, and X. S. Xie, “Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy,” J. Lipid Res. 44(11), 2202–2208 (2003).
[CrossRef] [PubMed]

Zhu, J.

Annu Rev Anal Chem (Palo Alto Calif)

C. L. Evans and X. S. Xie, “Coherent anti-stokes Raman scattering microscopy: chemical imaging for biology and medicine,” Annu Rev Anal Chem (Palo Alto Calif) 1(1), 883–909 (2008).
[CrossRef]

IEEE J. Sel. Top. Quantum Electron.

T. B. Huff, Y. Shi, Y. Fu, H. Wang, and J. X. Cheng, “Multimodal nonlinear optical microscopy and applications to central nervous system,” IEEE J. Sel. Top. Quantum Electron. 14(1), 4–9 (2008).
[CrossRef] [PubMed]

J. Biomed. Opt.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12(5), 054007 (2007).
[CrossRef] [PubMed]

J. T. C. Liu, M. J. Mandella, N. O. Loewke, H. Haeberle, H. Ra, W. Piyawattanametha, O. Solgaard, G. S. Kino, and C. H. Contag, “Micromirror-scanned dual-axis confocal microscope utilizing a gradient-index relay lens for image guidance during brain surgery,” J. Biomed. Opt. 15(2), 026029 (2010).
[CrossRef] [PubMed]

S. Tang, W. Jung, D. McCormick, T. Xie, J. Su, Y. C. Ahn, B. J. Tromberg, and Z. Chen, “Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning,” J. Biomed. Opt. 14(3), 034005 (2009).
[CrossRef] [PubMed]

J. Lipid Res.

X. L. Nan, J. X. Cheng, and X. S. Xie, “Vibrational imaging of lipid droplets in live fibroblast cells with coherent anti-Stokes Raman scattering microscopy,” J. Lipid Res. 44(11), 2202–2208 (2003).
[CrossRef] [PubMed]

J. Micro/ Nanolith MEMS MOEMS

M. Scholles, A. Bräuer, K. Frommhagen, Ch. Gerwig, H. Lakner, H. Schenk, and M. Schwarzenberg, “Ultracompact laser projection systems based on two-dimensional resonant microscanning mirrors,” J. Micro/ Nanolith MEMS MOEMS 7(2), 021001 (2008).
[CrossRef]

J. Microsc.

L. Fu and M. Gu, “Fibre-optic nonlinear optical microscopy and endoscopy,” J. Microsc. 226(3), 195–206 (2007).
[CrossRef] [PubMed]

E. Spiess, F. Bestvater, A. Heckel-Pompey, K. Toth, M. Hacker, G. Stobrawa, T. Feurer, C. Wotzlaw, U. Berchner-Pfannschmidt, T. Porwol, and H. Acker, “Two-photon excitation and emission spectra of the green fluorescent protein variants ECFP, EGFP and EYFP,” J. Microsc. 217(3), 200–204 (2005).
[CrossRef] [PubMed]

S. Murugkar, C. L. Evans, X. S. Xie, and H. Anis, “Chemically specific imaging of cryptosporidium oocysts using coherent anti-Stokes Raman scattering (CARS) microscopy,” J. Microsc. 233(2), 244–250 (2009).
[CrossRef] [PubMed]

Mol. Imaging

T. T. Le, C. W. Rehrer, T. B. Huff, M. B. Nichols, I. G. Camarillo, and J. X. Cheng, “Nonlinear optical imaging to evaluate the impact of obesity on mammary gland and tumor stroma,” Mol. Imaging 6(3), 205–211 (2007).
[PubMed]

Neuron

F. Helmchen, M. S. Fee, D. W. Tank, and W. Denk, “A miniature head-mounted two-photon microscope. high-resolution brain imaging in freely moving animals,” Neuron 31(6), 903–912 (2001).
[CrossRef] [PubMed]

Opt. Express

F. Légaré, C. L. Evans, F. Ganikhanov, and X. S. Xie, “Towards CARS Endoscopy,” Opt. Express 14(10), 4427–4432 (2006).
[CrossRef] [PubMed]

K. C. Maitland, H. J. Shin, H. Ra, D. Lee, O. Solgaard, and R. Richards-Kortum, “Single fiber confocal microscope with a two-axis gimbaled MEMS scanner for cellular imaging,” Opt. Express 14(19), 8604–8612 (2006).
[CrossRef] [PubMed]

A. Chong, J. Buckley, W. Renninger, and F. Wise, “All-normal-dispersion femtosecond fiber laser,” Opt. Express 14(21), 10095–10100 (2006).
[CrossRef] [PubMed]

C. L. Hoy, N. J. Durr, P. Chen, W. Piyawattanametha, H. Ra, O. Solgaard, and A. Ben-Yakar, “Miniaturized probe for femtosecond laser microsurgery and two-photon imaging,” Opt. Express 16(13), 9996–10005 (2008).
[CrossRef] [PubMed]

T. M. Liu, M. C. Chan, I. H. Chen, S. H. Chia, and C. K. Sun, “Miniaturized multiphoton microscope with a 24Hz frame-rate,” Opt. Express 16(14), 10501–10506 (2008).
[CrossRef] [PubMed]

H. Chen, H. Wang, M. N. Slipchenko, Y. Jung, Y. Shi, J. Zhu, K. K. Buhman, and J. X. Cheng, “A multimodal platform for nonlinear optical microscopy and microspectroscopy,” Opt. Express 17(3), 1282–1290 (2009).
[CrossRef] [PubMed]

S. Murugkar, C. Brideau, A. Ridsdale, M. Naji, P. K. Stys, and H. Anis, “Coherent anti-Stokes Raman scattering microscopy using photonic crystal fiber with two closely lying zero dispersion wavelengths,” Opt. Express 15(21), 14028–14037 (2007).
[CrossRef] [PubMed]

C. J. Engelbrecht, R. S. Johnston, E. J. Seibel, and F. Helmchen, “Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo,” Opt. Express 16(8), 5556–5564 (2008).
[CrossRef] [PubMed]

H. Bao, A. Boussioutas, R. Jeremy, S. Russell, and M. Gu, “Second harmonic generation imaging via nonlinear endomicroscopy,” Opt. Express 18(2), 1255–1260 (2010).
[CrossRef] [PubMed]

M. Balu, G. Liu, Z. Chen, B. J. Tromberg, and E. O. Potma, “Fiber delivered probe for efficient CARS imaging of tissues,” Opt. Express 18(3), 2380–2388 (2010).
[CrossRef] [PubMed]

Opt. Lett.

Proc. SPIE

M. Scholles, K. Frommhagen, Ch. Gerwig, J. Knobbe, H. Lakner, D. Schlebusch, M. Schwarzenberg, and U. Vogel, “Recent advancements in system design for miniaturized MEMS-based laser projectors,” Proc. SPIE 6911, 69110U (2008).
[CrossRef]

Other

I. Veilleux, M. Doucet, P. Coté, S. Verreault, M. Fortin, P. Paradis, S. Leclair, R. S. Da Costa, B. C. Wilson, E. Seibel, O. Mermut, and J. Cormier, “ Design and modeling of a prototype fiber scanning CARS endoscope,” Proc. SPIE, 7558, 75580D1 - 12 (2010).

B. G. Saar, R. S. Johnston, M. Roeffaers, E. J. Seibel and X. S. Xie, “CARS endoscopy,” presented at SPIE Photonics West, San Francisco, California, USA, 23 - 28 Jan. 2010.

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Figures (4)

Fig. 1
Fig. 1

Bench top miniaturized multimodal CARS microscope. Details of components are as follows: (2) Faraday isolator (3) Prism compressor (4) MEMS mirror (5) Miniature optics inside stainless steel tube (barrel) (6) Sample for imaging (7) microscope air objective (20x) for collecting nonlinear optical signal from sample (8) Lens for coupling light into collection fiber (9) Multimode fiber (10) Voltage Amplifier circuit to amplify electrical signal, for driving the MEMS mirror, provided by the Field Programmable Gate Array (FPGA).

Fig. 2
Fig. 2

(a) Ray-trace diagram of the optical beam as it enters from the right after the MEMS mirror and is relayed to the miniature focusing optics on the left, through a 5x beam expander assembly including the field lens. (b) Computer-aided-design of the barrel. (c) Photograph of the fully packaged barrel. A penny is included for size comparison.

Fig. 3
Fig. 3

(a) Transmission image in 720 nm light of the smallest bars in Group 7, element 6 on a USAF1951 target obtained with the miniature multimodal microscope. Part of the number ‘6’ can be seen in the top left side corner of the image. (b) TPEF image of a sample of 1 µm fluorescent microspheres on a thin #1 coverslip (c) CARS image of 20 µm polystyrene beads

Fig. 4
Fig. 4

Multimodal imaging using miniature CARS microscope (a) forward - CARS image at a Raman shift of 2845 cm−1 from label free myelin surrounding YFP labeled axons in fixed dorsal roots of mouse. (b) TPEF image of YFP labeled axons at excitation wavelength of 870 nm. (c) SHG image from unlabeled rat tail collagen.

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