Abstract

Histochemistry is a microscopy-based technology widely used to visualize the molecular distribution in biological tissue. Recent developments in label-free optical imaging has demonstrated the potential to replace the conventional histochemical labels/markers (fluorescent antibodies, organic dyes, nucleic acid probes, and other contrast agents) with diverse optical interactions to generate histochemical contrasts, allowing “virtual” histochemistry in three spatial dimensions without preparing a microscope slide (i.e. labor-intensive sample preparation). However, the histochemical information in a label-free optical image has often been rather limited due to the difficulty in simultaneously generating multiple histochemical contrasts with strict spatial co-registration. Here, in the first part (Part I) of this two-part series study, we develop a technique of slide-free virtual histochemistry based on label-free multimodal multiphoton microscopy, and simultaneously generate up to four histochemical contrasts from in vivo animal and ex vivo human tissue. To enable this functionality, we construct and demonstrate a robust fiber-based laser source for clinical translation and phenotype a wide variety of vital cells in unperturbed mammary tissue.

© 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

Full Article  |  PDF Article
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    [Crossref] [PubMed]

2018 (2)

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

2017 (6)

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

R. He, Y. Xu, L. Zhang, S. Ma, X. Wang, D. Ye, and M. Ji, “Dual-phase stimulated Raman scattering microscopy for real-time two-color imaging,” Optica 4(1), 44–47 (2017).
[Crossref]

W. Liu, S. H. Chia, H. Y. Chung, R. Greinert, F. X. Kärtner, and G. Chang, “Energetic ultrafast fiber laser sources tunable in 1030-1215 nm for deep tissue multi-photon microscopy,” Opt. Express 25(6), 6822–6831 (2017).
[Crossref] [PubMed]

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

2016 (2)

C. A. Alonzo, S. Karaliota, D. Pouli, Z. Liu, K. P. Karalis, and I. Georgakoudi, “Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function,” Sci. Rep. 6(1), 31012 (2016).
[Crossref] [PubMed]

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

2015 (2)

2014 (2)

A. C. Croce and G. Bottiroli, “Autofluorescence spectroscopy and imaging: a tool for biomedical research and diagnosis,” EJH 58, 4 (2014).

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

2013 (2)

H. Tu and S. A. Boppart, “Coherent fiber supercontinuum for biophotonics,” Laser Photonics Rev. 7(5), 628–645 (2013).
[Crossref] [PubMed]

C. Xu and F. W. Wise, “Recent advances in fibre lasers for nonlinear microscopy,” Nat. Photonics 7(11), 875–882 (2013).
[Crossref] [PubMed]

2011 (1)

2010 (2)

A. Yagci, W. Murk, J. Stronk, and G. Huszar, “Spermatozoa bound to solid state hyaluronic acid show chromatin structure with high DNA chain integrity: an acridine orange fluorescence study,” J. Androl. 31(6), 566–572 (2010).
[Crossref] [PubMed]

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

2007 (2)

J. E. Wagenseil and R. P. Mecham, “New insights into elastic fiber assembly,” Birth Defects Res. C Embryo Today 81(4), 229–240 (2007).
[Crossref] [PubMed]

N. Nishizawa and J. Takayanagi, “Octave spanning high-quality supercontinuum generation in all-fiber system,” J. Opt. Soc. Am. B 24(8), 1786–1792 (2007).
[Crossref]

2006 (1)

T. Sakimoto, M. I. Rosenblatt, and D. T. Azar, “Laser eye surgery for refractive errors,” Lancet 367(9520), 1432–1447 (2006).
[Crossref] [PubMed]

2003 (1)

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003).
[Crossref] [PubMed]

2002 (1)

S. Huang, A. A. Heikal, and W. W. Webb, “Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein,” Biophys. J. 82(5), 2811–2825 (2002).
[Crossref] [PubMed]

2001 (1)

Aft, R. L.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Ahsen, O. O.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Alfonso-García, A.

R. Datta, A. Alfonso-García, R. Cinco, and E. Gratton, “Fluorescence lifetime imaging of endogenous biomarker of oxidative stress,” Sci. Rep. 5(1), 9848 (2015).
[Crossref] [PubMed]

Alonzo, C. A.

C. A. Alonzo, S. Karaliota, D. Pouli, Z. Liu, K. P. Karalis, and I. Georgakoudi, “Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function,” Sci. Rep. 6(1), 31012 (2016).
[Crossref] [PubMed]

Aptel, F.

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

Azar, D. T.

T. Sakimoto, M. I. Rosenblatt, and D. T. Azar, “Laser eye surgery for refractive errors,” Lancet 367(9520), 1432–1447 (2006).
[Crossref] [PubMed]

Beaurepaire, E.

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

Boppart, S. A.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

H. Tu and S. A. Boppart, “Coherent fiber supercontinuum for biophotonics,” Laser Photonics Rev. 7(5), 628–645 (2013).
[Crossref] [PubMed]

S. You, Y. Sun, E. J. Chaney, Y. Zhao, J. Chen, S. A. Boppart, and H. Tu, “Slide-free virtual histochemistry (Part II): Detection of field cancerization,” Biomed. Opt. Express (to be published).

Bottiroli, G.

A. C. Croce and G. Bottiroli, “Autofluorescence spectroscopy and imaging: a tool for biomedical research and diagnosis,” EJH 58, 4 (2014).

Bower, A. J.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

Boyle, J.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Brooker, J. S.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Cable, A. E.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Cahill, L. C.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

Camelo-Piragua, S.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Cao, R.

Chaney, E. J.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

S. You, Y. Sun, E. J. Chaney, Y. Zhao, J. Chen, S. A. Boppart, and H. Tu, “Slide-free virtual histochemistry (Part II): Detection of field cancerization,” Biomed. Opt. Express (to be published).

Chang, G.

Chen, I. H.

Chen, J.

S. You, Y. Sun, E. J. Chaney, Y. Zhao, J. Chen, S. A. Boppart, and H. Tu, “Slide-free virtual histochemistry (Part II): Detection of field cancerization,” Biomed. Opt. Express (to be published).

Chen, P. C.

Chen, R.

Chen, Y.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Chia, S. H.

Christie, R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003).
[Crossref] [PubMed]

Chu, S. W.

Chung, H. Y.

Cinco, R.

R. Datta, A. Alfonso-García, R. Cinco, and E. Gratton, “Fluorescence lifetime imaging of endogenous biomarker of oxidative stress,” Sci. Rep. 5(1), 9848 (2015).
[Crossref] [PubMed]

Connolly, J. L.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Croce, A. C.

A. C. Croce and G. Bottiroli, “Autofluorescence spectroscopy and imaging: a tool for biomedical research and diagnosis,” EJH 58, 4 (2014).

Dagher, P. C.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Dantus, M.

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

Datta, R.

R. Datta, A. Alfonso-García, R. Cinco, and E. Gratton, “Fluorescence lifetime imaging of endogenous biomarker of oxidative stress,” Sci. Rep. 5(1), 9848 (2015).
[Crossref] [PubMed]

Day, R.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Deniset-Besseau, A.

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

Dixon, A. J.

Dunn, K. W.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Faulkner-Jones, B. E.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

Fisher-Hubbard, A.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Freudiger, C. W.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Fujimoto, J. G.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Garrard, M.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Garton, H. J. L.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Georgakoudi, I.

C. A. Alonzo, S. Karaliota, D. Pouli, Z. Liu, K. P. Karalis, and I. Georgakoudi, “Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function,” Sci. Rep. 6(1), 31012 (2016).
[Crossref] [PubMed]

Giacomelli, M. G.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

Glaser, A. K.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Gratton, E.

R. Datta, A. Alfonso-García, R. Cinco, and E. Gratton, “Fluorescence lifetime imaging of endogenous biomarker of oxidative stress,” Sci. Rep. 5(1), 9848 (2015).
[Crossref] [PubMed]

Greinert, R.

Hai, P.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Hato, T.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

He, R.

Heikal, A. A.

S. Huang, A. A. Heikal, and W. W. Webb, “Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein,” Biophys. J. 82(5), 2811–2825 (2002).
[Crossref] [PubMed]

Hervey-Jumper, S. L.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Heth, J. A.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Hollon, T. C.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Hooper, L. E.

Hossack, J. A.

Hu, S.

Huang, S.

S. Huang, A. A. Heikal, and W. W. Webb, “Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein,” Biophys. J. 82(5), 2811–2825 (2002).
[Crossref] [PubMed]

Huszar, G.

A. Yagci, W. Murk, J. Stronk, and G. Huszar, “Spermatozoa bound to solid state hyaluronic acid show chromatin structure with high DNA chain integrity: an acridine orange fluorescence study,” J. Androl. 31(6), 566–572 (2010).
[Crossref] [PubMed]

Hyman, B. T.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003).
[Crossref] [PubMed]

Ji, M.

Johnson, N. B.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Johnson, T. D.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Karaliota, S.

C. A. Alonzo, S. Karaliota, D. Pouli, Z. Liu, K. P. Karalis, and I. Georgakoudi, “Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function,” Sci. Rep. 6(1), 31012 (2016).
[Crossref] [PubMed]

Karalis, K. P.

C. A. Alonzo, S. Karaliota, D. Pouli, Z. Liu, K. P. Karalis, and I. Georgakoudi, “Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function,” Sci. Rep. 6(1), 31012 (2016).
[Crossref] [PubMed]

Kärtner, F. X.

Kennedy, M. J.

Kirk Shung, K.

Knight, J. C.

Lægsgaard, J.

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

Legeais, J. M.

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

Lewis, S.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Lieberman, A. P.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Lin, B. L.

Liu, J. T. C.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Liu, T. M.

Liu, W.

Liu, Y.

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

Liu, Y. Z.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

Liu, Z.

C. A. Alonzo, S. Karaliota, D. Pouli, Z. Liu, K. P. Karalis, and I. Georgakoudi, “Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function,” Sci. Rep. 6(1), 31012 (2016).
[Crossref] [PubMed]

Lyngsø, J.

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

Ma, S.

Maher, C. O.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Marjanovic, M.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

McCarty, E. F.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

McFadden, K. A.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Mecham, R. P.

J. E. Wagenseil and R. P. Mecham, “New insights into elastic fiber assembly,” Birth Defects Res. C Embryo Today 81(4), 229–240 (2007).
[Crossref] [PubMed]

Molitoris, B. A.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Mosley, P. J.

Muir, A. C.

Murk, W.

A. Yagci, W. Murk, J. Stronk, and G. Huszar, “Spermatozoa bound to solid state hyaluronic acid show chromatin structure with high DNA chain integrity: an acridine orange fluorescence study,” J. Androl. 31(6), 566–572 (2010).
[Crossref] [PubMed]

Nikitin, A. Y.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003).
[Crossref] [PubMed]

Niknafs, Y. S.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Ning, B.

Nishizawa, N.

Novack, D. V.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Olivier, N.

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

Orringer, D. A.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Pandian, B.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Plamann, K.

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

Pleitez, M. A.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Pouli, D.

C. A. Alonzo, S. Karaliota, D. Pouli, Z. Liu, K. P. Karalis, and I. Georgakoudi, “Two-photon excited fluorescence of intrinsic fluorophores enables label-free assessment of adipose tissue function,” Sci. Rep. 6(1), 31012 (2016).
[Crossref] [PubMed]

Pu, Y.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

Ramkissoon, S. H.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Reder, N. P.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Rosenblatt, M. I.

T. Sakimoto, M. I. Rosenblatt, and D. T. Azar, “Laser eye surgery for refractive errors,” Lancet 367(9520), 1432–1447 (2006).
[Crossref] [PubMed]

Sagher, O.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Sakimoto, T.

T. Sakimoto, M. I. Rosenblatt, and D. T. Azar, “Laser eye surgery for refractive errors,” Lancet 367(9520), 1432–1447 (2006).
[Crossref] [PubMed]

Sandoval, R. M.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Schanne-Klein, M. C.

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

Schmolze, D. B.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Sheikine, Y.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Shen, D.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Sinha, S.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

Snuderl, M.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Soetikno, B. T.

Stronk, J.

A. Yagci, W. Murk, J. Stronk, and G. Huszar, “Spermatozoa bound to solid state hyaluronic acid show chromatin structure with high DNA chain integrity: an acridine orange fluorescence study,” J. Androl. 31(6), 566–572 (2010).
[Crossref] [PubMed]

Sun, C. K.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
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S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, “Multimodal nonlinear spectral microscopy based on a femtosecond Cr:forsterite laser,” Opt. Lett. 26(23), 1909–1911 (2001).
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Sun, N.

Sun, Y.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

S. You, Y. Sun, E. J. Chaney, Y. Zhao, J. Chen, S. A. Boppart, and H. Tu, “Slide-free virtual histochemistry (Part II): Detection of field cancerization,” Biomed. Opt. Express (to be published).

Takayanagi, J.

Tao, Y. K.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Trautman, J. K.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

True, L. D.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Tu, H.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

H. Tu and S. A. Boppart, “Coherent fiber supercontinuum for biophotonics,” Laser Photonics Rev. 7(5), 628–645 (2013).
[Crossref] [PubMed]

S. You, Y. Sun, E. J. Chaney, Y. Zhao, J. Chen, S. A. Boppart, and H. Tu, “Slide-free virtual histochemistry (Part II): Detection of field cancerization,” Biomed. Opt. Express (to be published).

Turchinovich, D.

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

Vardeh, H.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

Venneti, S.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Wadsworth, W. J.

Wagenseil, J. E.

J. E. Wagenseil and R. P. Mecham, “New insights into elastic fiber assembly,” Birth Defects Res. C Embryo Today 81(4), 229–240 (2007).
[Crossref] [PubMed]

Wang, H. H.

Y. K. Tao, D. Shen, Y. Sheikine, O. O. Ahsen, H. H. Wang, D. B. Schmolze, N. B. Johnson, J. S. Brooker, A. E. Cable, J. L. Connolly, and J. G. Fujimoto, “Assessment of breast pathologies using nonlinear microscopy,” Proc. Natl. Acad. Sci. U.S.A. 111(43), 15304–15309 (2014).
[Crossref] [PubMed]

Wang, L. V.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Wang, X.

Wang, Y.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Webb, W. W.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003).
[Crossref] [PubMed]

S. Huang, A. A. Heikal, and W. W. Webb, “Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein,” Biophys. J. 82(5), 2811–2825 (2002).
[Crossref] [PubMed]

Wei, L.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Wiggins, R. C.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Wilkinson, D. A.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Williams, R. M.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003).
[Crossref] [PubMed]

Wilson, W. L.

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

Winfree, S.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Wise, F. W.

C. Xu and F. W. Wise, “Recent advances in fibre lasers for nonlinear microscopy,” Nat. Photonics 7(11), 875–882 (2013).
[Crossref] [PubMed]

Wong, T. T. W.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Xie, X. S.

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

Xu, B.

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

Xu, C.

C. Xu and F. W. Wise, “Recent advances in fibre lasers for nonlinear microscopy,” Nat. Photonics 7(11), 875–882 (2013).
[Crossref] [PubMed]

Xu, Y.

Yagci, A.

A. Yagci, W. Murk, J. Stronk, and G. Huszar, “Spermatozoa bound to solid state hyaluronic acid show chromatin structure with high DNA chain integrity: an acridine orange fluorescence study,” J. Androl. 31(6), 566–572 (2010).
[Crossref] [PubMed]

Ye, D.

Yin, C.

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
[Crossref] [PubMed]

Yoder, M. C.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Yoshitake, T.

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

You, S.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

S. You, Y. Sun, E. J. Chaney, Y. Zhao, J. Chen, S. A. Boppart, and H. Tu, “Slide-free virtual histochemistry (Part II): Detection of field cancerization,” Biomed. Opt. Express (to be published).

Zhang, C.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Zhang, L.

Zhang, R.

T. T. W. Wong, R. Zhang, P. Hai, C. Zhang, M. A. Pleitez, R. L. Aft, D. V. Novack, and L. V. Wang, “Fast label-free multilayered histology-like imaging of human breast cancer by photoacoustic microscopy,” Sci. Adv. 3(5), e1602168 (2017).
[Crossref] [PubMed]

Zhao, Y.

S. You, H. Tu, E. J. Chaney, Y. Sun, Y. Zhao, A. J. Bower, Y. Z. Liu, M. Marjanovic, S. Sinha, Y. Pu, and S. A. Boppart, “Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy,” Nat. Commun. 9(1), 2125 (2018).
[Crossref] [PubMed]

H. Tu, Y. Liu, D. Turchinovich, M. Marjanovic, J. Lyngsø, J. Lægsgaard, E. J. Chaney, Y. Zhao, S. You, W. L. Wilson, B. Xu, M. Dantus, and S. A. Boppart, “Stain-free histopathology by programmable supercontinuum pulses,” Nat. Photonics 10(8), 534–540 (2016).
[Crossref] [PubMed]

S. You, Y. Sun, E. J. Chaney, Y. Zhao, J. Chen, S. A. Boppart, and H. Tu, “Slide-free virtual histochemistry (Part II): Detection of field cancerization,” Biomed. Opt. Express (to be published).

Zheng, Y.

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

Zhou, Q.

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. U.S.A. 100(12), 7075–7080 (2003).
[Crossref] [PubMed]

Biophys. J. (1)

S. Huang, A. A. Heikal, and W. W. Webb, “Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein,” Biophys. J. 82(5), 2811–2825 (2002).
[Crossref] [PubMed]

Birth Defects Res. C Embryo Today (1)

J. E. Wagenseil and R. P. Mecham, “New insights into elastic fiber assembly,” Birth Defects Res. C Embryo Today 81(4), 229–240 (2007).
[Crossref] [PubMed]

EJH (1)

A. C. Croce and G. Bottiroli, “Autofluorescence spectroscopy and imaging: a tool for biomedical research and diagnosis,” EJH 58, 4 (2014).

Invest. Ophthalmol. Vis. Sci. (1)

F. Aptel, N. Olivier, A. Deniset-Besseau, J. M. Legeais, K. Plamann, M. C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[Crossref] [PubMed]

J. Am. Soc. Nephrol. (1)

T. Hato, S. Winfree, R. Day, R. M. Sandoval, B. A. Molitoris, M. C. Yoder, R. C. Wiggins, Y. Zheng, K. W. Dunn, and P. C. Dagher, “Two-photon intravital imaging fluorescence lifetime imaging of the kidney reveals cell-type specific metabolic signatures,” J. Am. Soc. Nephrol. 28(8), 2420–2430 (2017).
[Crossref] [PubMed]

J. Androl. (1)

A. Yagci, W. Murk, J. Stronk, and G. Huszar, “Spermatozoa bound to solid state hyaluronic acid show chromatin structure with high DNA chain integrity: an acridine orange fluorescence study,” J. Androl. 31(6), 566–572 (2010).
[Crossref] [PubMed]

J. Opt. Soc. Am. B (1)

Lab. Invest. (1)

L. C. Cahill, M. G. Giacomelli, T. Yoshitake, H. Vardeh, B. E. Faulkner-Jones, J. L. Connolly, C. K. Sun, and J. G. Fujimoto, “Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope,” Lab. Invest. 98(1), 150–160 (2018).
[Crossref] [PubMed]

Lancet (1)

T. Sakimoto, M. I. Rosenblatt, and D. T. Azar, “Laser eye surgery for refractive errors,” Lancet 367(9520), 1432–1447 (2006).
[Crossref] [PubMed]

Laser Photonics Rev. (1)

H. Tu and S. A. Boppart, “Coherent fiber supercontinuum for biophotonics,” Laser Photonics Rev. 7(5), 628–645 (2013).
[Crossref] [PubMed]

Nat. Biomed. Eng. (2)

D. A. Orringer, B. Pandian, Y. S. Niknafs, T. C. Hollon, J. Boyle, S. Lewis, M. Garrard, S. L. Hervey-Jumper, H. J. L. Garton, C. O. Maher, J. A. Heth, O. Sagher, D. A. Wilkinson, M. Snuderl, S. Venneti, S. H. Ramkissoon, K. A. McFadden, A. Fisher-Hubbard, A. P. Lieberman, T. D. Johnson, X. S. Xie, J. K. Trautman, C. W. Freudiger, and S. Camelo-Piragua, “Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy,” Nat. Biomed. Eng. 1(2), 0027 (2017).
[Crossref] [PubMed]

A. K. Glaser, N. P. Reder, Y. Chen, E. F. McCarty, C. Yin, L. Wei, Y. Wang, L. D. True, and J. T. C. Liu, “Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens,” Nat. Biomed. Eng. 1(7), 0084 (2017).
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Supplementary Material (4)

NameDescription
» Visualization 1       Volumetric image corresponding to Fig. 3b.
» Visualization 2       Volumetric image corresponding to Fig. 3c.
» Visualization 3       Volumetric image corresponding to Fig. 3g.
» Visualization 4       Volumetric image corresponding to Fig. 3t.

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Figures (5)

Fig. 1
Fig. 1 Slide-free virtual histochemistry of fresh untreated tissue using a fiber laser-induced supercontinuum source. (a) Schematic of tetra-modal inverted microscope, with specific sets of dichroic mirrors, optical filters, photomultipliers (PMTs) (upper panel) that dictate 4 optical markers/colors (lower panel). (b) Upper panel: transmission bands of detection channels against emission spectra of NADH and FAD; lower panel: signal generation from an FAD solution, an NADH solution, and saline (control), with intensity computed by averaging the pixel values of the image in corresponding channels. (c) Comparison of supercontinuum source generated by a solid-state laser (upper panel) and the fiber laser in this study (lower panel). (d) Images of epithelial cells in a mouse kidney showing combined 2PAF-3PAF contrasts of the tetra-modal imaging (left panel), 2PAF contrast alone (middle panel), and 2PAF contrast at reprogrammed (920-nm) excitation detected by the SHG channel of the tetra-modal imaging. (e) Images of collagen and elastin fibers in human breast tissue showing combined 2PAF-3PAF contrasts of the tetra-modal imaging (left panel), 2PAF contrast alone (middle panel), and 2PAF contrast at reprogrammed (920-nm) excitation detected by the SHG channel of the tetra-modal imaging. (f) Tetra-modal images of human breast tissue showing cyan-colored lipid vacuoles of adipocytes (arrows) and magenta-colored cytoplasm/mitochondria of certain cells (arrowheads) before acridine orange labeling (upper panel) and after the labeling (lower panels). An average focusing power of ~20 mW was used on the tissue. Scale bars: 50 µm.
Fig. 2
Fig. 2 Comparison of the tetra-modal images of normal mammary tissue before and after acridine orange labeling. Although labeling of nuclei is validated with green contrast (see Fig. 1(f)), many cells exhibit increased production of FAD (yellow contrast) associated with the labeling. Scale bars: 100 µm.
Fig. 3
Fig. 3 Optical phenotyping of related rat mammary and human breast cells in different microenvironments by slide-free virtual histochemistry. (a) Rat immune cells in stroma. (b) Rat endothelial cells in a developing blood vessel (Visualization 1). (c) Rat tubular structure (Visualization 2). (d) Rat tumor cells with a largely cyan-colored cytoplasm. (e) Rat tumor structure. (f) Normal rat stromal cells. (g) Rat fibroblasts aligned with a blood vessel (Visualization 3). (h) Cyan-colored rat cancer-associated cells. (i) Magenta-colored rat cancer-associated cells (arrow) near mammary ducts (arrowhead). (j) Rat tumor cells with a magenta-colored cell body. (k) Normal human stromal cells. (l) Human fibroblasts aligned with a blood vessel. (m) Cyan-colored human cancer-associated cells. (n) Magenta-colored human cancer-associated cells (arrow) near mammary ducts. (o) Human tumor cells with a magenta-colored cytoplasm. (p) Human immune cells in stroma. (q) A plausible developing human blood vessel. (r) Human tubular structure. (s) Human tumor cells with a largely cyan-colored cytoplasm. (t) Human tumor structure (Visualization 4). Scale bars: 50 µm.
Fig. 4
Fig. 4 Tetra-modal images of normal mammary tissue and tumors from different rats. Tumors from three different rats exhibit large heterogeneity in optical phenotype. Dashed box 1 is also plotted in Fig. 3(j). Scale bars: 100 µm.
Fig. 5
Fig. 5 Tetra-modal images of normal breast tissue and tumors from different human subjects. Tumors from three different patients exhibit large heterogeneity in optical phenotype. Dashed boxes 1 and 2 are also plotted in Figs. 3(o) and 3(s), respectively. Scale bars: 100 µm. Dash box 3 shows magenta-colored stromal cells (arrows).

Tables (2)

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Table 1 Some coherent fiber supercontinuum sources for general multiphoton microscopy.

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Table 2 Classified vital stromal or tumor cells in mammary tissue with distinct optical phenotypes.

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