Stain-free histopathology by programmable supercontinuum pulses.
The preparation, staining, visualization, and interpretation of histological images of tissue is well-accepted as the gold standard process for the diagnosis of disease. These methods were developed historically, and are used ubiquitously in pathology, despite being highly time and labor intensive. Here we introduce a unique optical imaging platform and methodology for label-free multimodal multiphoton microscopy that uses a novel photonic crystal fiber source to generate tailored chemical contrast based on programmable supercontinuum pulses. We demonstrate collection of optical signatures of the tumor microenvironment, including evidence of mesoscopic biological organization, tumor cell migration, and (lymph-)angiogenesis collected directly from freshex vivomammary tissue. Acquisition of these optical signatures and other cellular or extracellular features, which are largely absent from histologically processed and stained tissue, combined with an adaptable platform for optical alignment-free programmable-contrast imaging, offers the potential to translate stain-free molecular histopathology into routine clinical use.
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Haohua
H
Tu
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Yuan
Y
Liu
Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA, Max Planck Institute for Polymer Research, Ackermannweg 10, Mainz, 55128, Germany.
Dmitry
D
Turchinovich
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA, Max Planck Institute for Polymer Research, Ackermannweg 10, Mainz, 55128, Germany.
Marina
M
Marjanovic
NKT Photonics A/S, Blokken 84, 3460 Birkerød, Denmark.
Jens
J
Lyngsø
DTU Fotonik, Technical University of Denmark, Ørsteds Plads 343, 2800 Lyngby, Denmark.
Jesper
J
Lægsgaard
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Eric J
EJ
Chaney
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Youbo
Y
Zhao
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA, Max Planck Institute for Polymer Research, Ackermannweg 10, Mainz, 55128, Germany.
Sixian
S
You
Department of Materials Science and Engineering, Frederick Seitz Materials Research Laboratory, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
William L
WL
Wilson
Biophotonic Solutions Inc., East Lansing, Michigan 48823, USA.
Bingwei
B
Xu
Department of Chemistry and Department of Physics, Michigan State University, East Lansing, Michigan 48824, USA.
Marcos
M
Dantus
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA, Max Planck Institute for Polymer Research, Ackermannweg 10, Mainz, 55128, Germany; Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; College of Medicine, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Stephen A
SA
Boppart
Nature. 1983 Dec 15-21;306(5944):658-61
6318112
Science. 1990 Apr 6;248(4951):73-6
2321027
Ann Diagn Pathol. 2007 Apr;11(2):137-41
17349575
Proc Natl Acad Sci U S A. 2007 Dec 4;104(49):19494-9
18042710
BMC Cancer. 2009 Jan 30;9:42
19183472
J Lightwave Technol. 2015 May 1;33(9):1814-1820
26166939
Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7075-80
12756303
Sci Transl Med. 2013 Sep 4;5(201):201ra119
24005159
Nature. 2009 Oct 22;461(7267):1105-9
19847261
Science. 1991 Nov 22;254(5035):1178-81
1957169
Cell. 2011 Mar 4;144(5):646-74
21376230
Nat Rev Cancer. 2002 Aug;2(8):573-83
12154350
J Surg Res. 1992 Mar;52(3):205-8
1311399
Opt Express. 2009 Feb 16;17(4):2984-96
19219203
Sci Transl Med. 2011 Feb 23;3(71):71ra15
21346168
Nat Rev Mol Cell Biol. 2009 Jul;10(7):445-57
19546857
Lab Invest. 2000 Feb;80(2):221-31
10701691
Opt Lett. 2004 Apr 1;29(7):775-7
15072388
Cancer Res. 2010 Dec 1;70(23):9562-9
21098699
IEEE J Sel Top Quantum Electron. 2012 May;18(3):null
24187481
J Am Acad Dermatol. 2002 Dec;47(6):869-74
12451371
Nat Med. 2011 Nov 07;17(11):1359-70
22064426
BMC Med. 2006 Dec 26;4(1):38
17190588
Proc Natl Acad Sci U S A. 2014 Oct 28;111(43):15304-9
25313045
Nature. 2000 Sep 14;407(6801):249-57
11001068
BMC Med. 2008 Apr 28;6:11
18442412
Opt Lett. 2007 Sep 15;32(18):2641-3
17873920
Nat Methods. 2012 Jul 08;9(8):815-8
22772730
J Biol Chem. 1979 Jun 10;254(11):4764-71
220260
Ann Diagn Pathol. 2008 Dec;12(6):387-96
18995201
Science. 2009 May 22;324(5930):1029-33
19460998
Endocrinology. 1982 Aug;111(2):584-8
7094887
Nat Biotechnol. 2006 Jul;24(7):848-51
16823374
Cell. 2009 Nov 25;139(5):891-906
19931152
Proc Natl Acad Sci U S A. 2015 Sep 15;112(37):11624-9
26324899
Nat Biotechnol. 2003 Nov;21(11):1369-77
14595365
Nature. 1980 Dec 11;288(5791):551-6
6160403
Nat Rev Cancer. 2007 Oct;7(10):763-77
17882277
Ann Diagn Pathol. 2009 Aug;13(4):246-56
19608083