Sponsored by SPIE Cosponsored by Becker and Hickl (Germany) • Carl Zeiss • Chroma Technology Corporation (United States) • Coherent Inc. • Semrock Inc. (United States) • ISS, Inc. • JenLab GmbH (Germany) • Leica Mircosystems • Spectra-Physics®, a Newport Corporation Brand (United States) • Princeton Instruments • Veroptics (United States) Published by SPIE Volume 9329 Proceedings of SPIE, 1605-7422, V. 9329 SPIE is an international society advancing an interdisciplinary approach to the science and application of light. The papers included in this volume were part of the technical conference cited on the cover and title page. Papers were selected and subject to review by the editors and conference program committee. Some conference presentations may not be available for publication. The papers published in these proceedings reflect the work and thoughts of the authors and are published herein as submitted. The publisher is not responsible for the validity of the information or for any outcomes resulting from reliance thereon. Please use the following format to cite material from this book: Author(s), “Title of Paper,” in Multiphoton Microscopy in the Biomedical Sciences XV, edited by Ammasi Periasamy, Peter T. C. So, Karsten König, Proceedings of SPIE Vol. 9329 (SPIE, Bellingham, WA, 2015) Article CID Number. ISSN: 1605-7422 ISBN: 9781628414196 Published by SPIE P.O. Box 10, Bellingham, Washington 98227-0010 USA Telephone +1 360 676 3290 (Pacific Time) · Fax +1 360 647 1445 SPIE.org Copyright © 2015, Society of Photo-Optical Instrumentation Engineers. Copying of material in this book for internal or personal use, or for the internal or personal use of specific clients, beyond the fair use provisions granted by the U.S. Copyright Law is authorized by SPIE subject to payment of copying fees. The Transactional Reporting Service base fee for this volume is $18.00 per article (or portion thereof), which should be paid directly to the Copyright Clearance Center (CCC), 222 Rosewood Drive, Danvers, MA 01923. Payment may also be made electronically through CCC Online at copyright.com. Other copying for republication, resale, advertising or promotion, or any form of systematic or multiple reproduction of any material in this book is prohibited except with permission in writing from the publisher. The CCC fee code is 1605-7422/15/$18.00. Printed in the United States of America. Publication of record for individual papers is online in the SPIE Digital Library. SPIEDigitalLibrary.org Paper Numbering: Proceedings of SPIE follow an e-First publication model, with papers published first online and then in print. Papers are published as they are submitted and meet publication criteria. A unique citation identifier (CID) number is assigned to each article at the time of the first publication. Utilization of CIDs allows articles to be fully citable as soon as they are published online, and connects the same identifier to all online, print, and electronic versions of the publication. SPIE uses a six-digit CID article numbering system in which: ▪ The first four digits correspond to the SPIE volume number. ▪ The last two digits indicate publication order within the volume using a Base 36 numbering system employing both numerals and letters. These two-number sets start with 00, 01, 02, 03, 04, 05, 06, 07, 08, 09, 0A, 0B … 0Z, followed by 10-1Z, 20-2Z, etc.
The CID Number appears on each page of the manuscript. The complete citation is used on the first page, and an abbreviated version on subsequent pages. AuthorsNumbers in the index correspond to the last two digits of the six-digit citation identifier (CID) article numbering system used in Proceedings of SPIE. The first four digits reflect the volume number. Base 36 numbering is employed for the last two digits and indicates the order of articles within the volume. Numbers start with 00, 01, 02, 03, 04, 05, 06, 07, 08, 09, 0A, 0B...0Z, followed by 10-1Z, 20-2Z, etc. Aimé, C., 2M Allegra Mascaro, A. L., 03 Avti, Pramod, 29 Baldacchini, T., 0V Bancelin, S., 2M Barbano, E. C., 2L Batista, Ana, 0E, 0U, 2P Beaurepaire, Emmanuel, 0Z Beeby, Andrew, 39 Beeson, K., 2O Bélanger, Samuel, 29 Bisinger, D., 0C Blab, Gerhard A., 0H Borisov, Sergei, 0B Börsch, Michael, 0A Brehm, Bernhard, 19 Breunig, Hans Georg, 0E, 0U, 1N, 2P Breymayer, J., 0C Brideau, Craig, 2U, 2X Brinchman, Jan E., 1C Brown-Steinke, Kathleen, 0G Bruncko, J., 0D Bub, G., 36 Bückle, Rainer, 0Q Burton, R. A. B., 36 Campagnola, Paul J., 2A Campbell, Kirby R., 2A Castonguay, Alexandre, 29 Chandrappa, Dayananda, 1J Chen, Jerry L. 2C Chen, Xueqin, 2H Chen, Xun, 2W Chorvat, D., Jr., 0D Cicchi, Riccardo, 19 Coradin, T., 2M Corbett, A. D., 36 Costantini, I., 03 Coura, J. A., 2L Dall, Christopher P., 0P Dana, Hod, 28 Deb, Sanghamitra, 0P Degan, Simone, 0P Dietzek, Benjamin, 19 DiMarzio, Charles A., 2I Dmitriev, Ruslan I., 0B Duport, François, 2G Düser, Monika G., 0A Ehmke, Tobias, 12 Emplit, Philippe, 2G Fereidouni, Farzad, 0H Finnøy, Andreas, 1C Formanek, Florian, 2H Franke, T., 0V Gainey, Christina S., 0P Galey, Jean-Baptiste, 2H Gao, Jian, 0N Garrett, N. L., 22 Gasecka, Alicja, 2H Gerlach, Nicole, 0Q Gerritsen, Hans C., 0H Gladine, C., 0D Godfrey, L., 22 Goldman, Serge, 2G Gorza, Simon-Pierre, 2G Gu, Zetong, 2I Gusachenko, I., 2M Hakulinen, T., 0V Hase, Eiji, 2Q Heinrich, Ulrike, 0Q Heisterkamp, Alexander, 12 Heitkamp, Thomas, 0A Helmchen, Fritjof, 2C Hirosawa, Kenichi, 2N Hirvonen, Liisa M., 39 Horilova, J., 0D Huang, Zhiwei, 0T, 1P Isobe, Keisuke, 2N Kalinina, S., 0C Kannari, Fumihiko, 2N Klein, J., 0V Knebl, Andreas, 12 Knott, G. W., 03 König, Karsten, 0E, 0Q, 0U, 1N, 2P, 33 Kowalczuk, L., 2M Krüppel, Roland, 2C Lai, Zhenhua, 2I Lajdova, I., 0D Lalatsa, A., 22 Lamb, Erin S., 20 Latour, G., 2M Lattermann, Annika, 19 Lei, Ming, 2W Le Marois, Alix, 38 Lesage, Frédéric, 29 Li, Runze, 2W Lilledahl, Magnus, 1C Lin, Danying, 0N Lin, Jian, 0T, 1P Luo, Yi, 0O Maco, B., 03 Mahou, Pierre, 0Z Marcek Chorvatova, A., 0D Mateasik, A., 0D Matthäus, Christian, 19 Mayblum, Tom, 28 Meyer, Tobias, 19 Midorikawa, Katsumi, 2N Miguez, M. L., 2L Misoguti, L., 2L Moeini, Mohammad, 29 Moger, Julian, 1J, 22 Morgado, António Miguel, 0E Nguyen, Anh Dung, 2G Niu, Hanben, 0N Olderøy, Magnus, 1C Olstad, Kristin, 1C Owen, Dylan M., 38 Ozeki, Yasuyuki, 1T Palmer, Lisa A., 0G Papkovsky, Dmitri B., 0B Parilov, E., 2O Pavone, Francesco S., 03, 19 Pei, Hanzhang, 20 Peng, Tong, 2W Peng, Xiao, 0N Periasamy, Ammasi, 0G, 3A Petrášek, Zdenek, 39 Pietruszka, Anna, 0Q Poon, Kelvin, 2U Popp, Jürgen, 19 Potasek, M., 2O Pouliot, Philippe, 29 Qu, Jianan Y., 0O Qu, Junle, 0N Rehman, Shagufta, 0G Reitsma, Keimpe, 0H Rigneault, Hervé, 2H Rowlands, Christopher J., 27 Rück, A., 0C Sacconi, L., 03 Sato, Katsuya, 2Q Schanne-Klein, M.-C., 2M Schatzlein, A., 22 Schejter, Adi, 28 Serrano, D. R., 22 Shah, Amy T., 2B Shoham, Shy, 28 Silvestri, L., 03 Skala, Melissa C., 2B Smirnoff, Nicholas, 1J So, Peter T. C., 27 Song, Qiyuan, 2N Starke, Ilka, 0A Stys, Peter, 2U, 2X Su, Bertram, 0A Suhling, Klaus, 38, 39 Sun, Qiqi, 0O Sun, Yuansheng, 3A Supatto, Willy, 0Z Svindrych, Zdenek, 3A Tabatabaei, Maryam S., 29 Tameze-Rivas, Yasmine, 0P Teh, Sengkhoon, 0T Teplicky, T., 0D Tilbury, Karissa B., 2A Uchegbu, I. F., 22 Uchugonova, Aisada, 0E, 0U, 2P Vanholsbeeck, Frédérique, 2G Van Horn, Mark, 2W Vermot, Julien, 0Z Voigt, Fabian F, 2C Wallrabe, Horst, 3A Wang, Chun-Chin, 1J Wang, Qi, 0N Wang, Zi, 0T, 1P Warren, Warren S., 0P Weinigel, Martin, 0Q, 1N Wilson, Jesse W., 0P Wilson, T., 36 Wise, Frank W., 20 Wu, Di, 2W Xue, Yi, 27 Yan, Wei, 0N Yang, Yanlong, 2W Yao, Baoli, 2W Yasui, Takeshi, 2Q Ye, Tong, 0N, 2W Yin, Jihao, 2I Zadoyan, R., 0V Zareinia, Kourosh, 2X Zarrabi, Nawid, 0A Zeng, Yan, 0O Zhang, Jennifer Y., 0P Zhang, Wei, 0O Zhang, Xi, 2I Zheng, Wei, 0O Zheng, Wei, 0T, 1P Zhou, Jie, 0N Zhou, Xing, 2W Zilio, S. C., 2L
Conference CommitteeSymposium Chairs James G. Fujimoto, Massachusetts Institute of Technology (United States) R. Rox Anderson, Wellman Center for Photomedicine, Massachusetts General Hospital (United States)and Harvard School of Medicine (United States)
Program Track Chairs Ammasi Periasamy, University of Virginia (United States) Daniel L. Farkas University of Southern California (United States)
Conference Chairs Ammasi Periasamy, University of Virginia (United States) Peter T. C. So, Massachusetts Institute of Technology (United States) Karsten König, Universität des Saarlandes (Germany)
Conference Program Committee Wolfgang Becker, Becker & Hickl GmbH (Germany) Alberto Diaspro, Istituto Italiano di Tecnologia (Italy) Chen-Yuan Dong, National Taiwan University (Taiwan) Kevin W. Eliceiri, University of Wisconsin-Madison (United States) Scott Fraser, California Institute of Technology (United States) Paul M. W. French, Imperial College London (United Kingdom) Hans C. Gerritsen, Universteit Utrecht (Netherlands) Enrico Gratton, University of California, Irvine (United States) Min Gu, Swinburne University of Technology (Australia) Stefan W. Hell, Max-Planck-Institut für Biophysikalische Chemie (Germany) Paul J. Campagnola, University of Wisconsin-Madison (United States) Satoshi Kawata, Osaka University (Japan) Fu-Jen Kao, National Yang-Ming University (Taiwan) Arnd K. Krueger, Spectra-Physics®, a Newport Corporation Brand (United States) Joseph R. Lakowicz, University of Maryland School of Medicine (United States) Steve M. McDonald, Coherent, Inc. (United States) Angelika C. Rueck, Universtät Ulm (Germany) Junle Qu, Shenzhen University (China) Steven S. Vogel, National Institutes of Health (United States) Paul W. Wiseman, McGill University (Canada) X. Sunney Xie, Harvard University (United States) Bernhard Zimmermann, Carl Zeiss Jena GmbH (Germany) Warren R. Zipfel, Cornell University (United States) Chris Xu, Cornell University (United States)
Session Chairs 1 Keynote Session
2 FLIM/FRET/FCS I
3 FLIM/FRET/FCS II
4 Technology Development and Application I
5 Technology Development and Application II
Aisada Uchugonova, Universität des Saarlandes (Germany) JenLab Young Investigator Award Papers Presentation Ammasi Periasamy, University of Virginia (United States)
6 Technology Development and Application III
7 SHG/THG Microscopy I
Chen-Yuan Dong, National Taiwan University (Taiwan) Award Presentation Ammasi Periasamy, University of Virginia (United States)
8 SHG/THG Microscopy II
9 Biomedical Applications of Coherent Raman I
10 Biomedical Applications of Coherent Raman II
11 Coherent Raman Technical Development I
12 Coherent Raman Technical Development II
Dan Fu, Harvard University (United States) Posters-Sunday Holly Aaron, University of California, Berkeley (United States) Steven S. Vogel, National Institutes of Health (United States) Jesse W. Wilson, Duke University (United States) Kevin W. Eliceiri, University of Wisconsin-Madison (United States)
IntroductionMultiphoton microscopy has been established as the 3D imaging method of choice for studying biomedical specimens from single cells to whole animals with sub-micron resolution. Two decades have passed since the realization of two-photon laser scanning microscopy. The ever-expanding scope of applications and the continuing instrumental innovations require a forum where new ideas can be exchanged and presented. Our conference at the SPIE BiOS 2015 meeting continues to address this need. This was the 15th year of this conference and we started our conference with three keynote lectures from leaders in the field: Drs. Paras N. Prasad, Univ. at Buffalo (United States) [9329-1], Francesco S. Pavone, European Lab. for Non-linear Spectroscopy (Italy) [9329-2], and Wolfgang Becker, Becker & Hickl GmbH (Germany) [9329-3]. For the 4th year in a row, the conference was extremely pleased to have the JenLab Young Investigator Award in addition to our regular poster awards. This award was donated by Dr. Karsten König, President and Founder of JenLab GmbH (Germany). The award selection committee included Drs. Arnd Krueger, NewPort-Spectra Physics (United States); Conor Evans, Harvard Univ. (United States); Alberto Diaspro, Istituto Italiana di Tecnologia (Italy); and two of the conference chairs, Karsten König and Ammasi Periasamy. The selection process included the abstract, manuscript, and poster presentation. Two finalists were selected for oral presentation after their poster presentation. The two finalists were (1) Mr. Kirby R. Campbell from Univ. of Wisconsin-Madison (United States), for “Determination of the spectral dependence of reduced scattering and quantitative SHG imaging for detection of fibrillary changes in ovarian cancer” [9329-79] and (2) Ms. Yi Xue from Massachusetts Institute of Technology, Boston, Massachusetts (United States), for “Parallel and flexible imaging using two-photon RESOLFT microscopy with spatial light modulator control” [9329-76]. Ms. Yi Xue from Massachusetts Institute of Technology, Boston, Massachusetts (United States) was selected as the winner of the JenLab Young Investigator Award 2015. For 15 years in a row, the conference organized poster awards for the students and postdoctoral fellows. The poster award was donated by all the conference sponsors including: Becker & Hickl, Chroma Technology, Coherent, ISS, Spectra-Physics, Princeton Instruments, Leica Microsystems, Semrock (IDEX), Veroptics and Carl Zeiss. The 4 poster award winners were: 1. Charles-André Couture, Institut National de la Recherche Scientifique (Canada): “Interferometric second harmonic generation imaging of biological tissues” [9329-75] 2. Dapeng Zhang, The Chinese Univ. of Hong Kong (Hong Kong, China): “Multi-photon laser scanning omnidirectional imaging with tunable frame rate” [9329-89] 3. Joe T. Sharick, Vanderbilt Univ. (United States): “Monitoring metabolic enzyme activity in cells with fluorescence lifetime imaging of NAD(P)H” [9329-107] 4. Alexander D. Corbett, Univ. of Oxford (United Kingdom): “Imaging cardiomyocytes in intact tissue with a remote focusing microscope” [9329-114]
Some of the most valuable contributions in this volume are articles written by highly experienced practitioners of multiphoton microscopy. They have enumerated the most important considerations in designing multi-photon microscopes and imaging experiments. Further, updates on the state-of-the-art commercial multiphoton microscope systems are presented. This volume also includes articles describing some recent advances in major multiphoton microscope components and applications including laser light sources, ultrafast optics, filters, FRET, FLIM, FCS, Raman, CARS, SRS and CRS microscopy and spectroscopy, single molecule, super-resolution imaging, endoscopy, and various scientific and clinical applications. On a personal note, the conference chairs are grateful for the participation of all authors and session chairs; and acknowledge the innovation-driven manufacturers and sponsors of this conference (Becker & Hickl, Chroma Technology, Coherent, ISS, Spectra-Physics, Princeton Instruments, Leica Microsystems, Semrock (IDEX), Veroptics, Carl Zeiss, and JenLab) for their enthusiastic support in organizing this conference successfully for the last 15 years. We look forward to other exciting conferences in the second decade and welcome your continued participation and support. Ammasi Periasamy Karsten König Peter T. C. So Congratulations to the 2014 Nobel Laureates in Chemistry One personal note to the Fluorescence Microscopy Community: We were all delighted and congratulated the 2014 Nobel Laureates in Chemistry at the Photonics West 2015 BiOS in a special plenary session on Nobel award on 8 February 2015. This is the first time the SPIE organization celebrated the Nobel award in a special plenary session. Three scientists’ shared the 2014 Nobel award for their achievements in super resolution microscopy: Dr. Eric Betzig from Janelia Farm, Howard Hughes Medical Institute, Ashburn, Virginia (United States), for his achievement in Photoactivated Localization Microscopy (PALM). Dr. Stefan W. Hell from Max-Planck-Institut für biophysikalische Chemie (Germany), for his achievement in Stimulated Emission Depletion (STED) microscopy. Dr. William E. Moerner (WE) from Stanford Univ., California (United States), for his achievement in single molecule detection. All the ‘buzz’ of the Nobel award was about that “Living Room”, where Eric Betzig built the PALM microscopy imaging system in his friend’s (Dr. Harald Hess) home. Eric Betzig dedicated his Nobel lecture about super-resolution optics to “all people taking risks but in the end failed”. William Moerner mentioned that a single molecule is extremely small, which he was able to detect using various techniques. Stefan Hell also discovered in his journey to the award that it is possible to overcome the generally accepted resolution barrier. There is nothing that is impossible in science. All three have presented other scientific work in this conference at various times. Stefan Hell for example gave two memorable keynote lectures at this multiphoton conference during 2005, “Fluorescence nanoscopy through reversible optically saturable transitions” [5700-01], and again in 2010, “Nanoscopy with focused light” [7569-02]. The editors wish to congratulate the 2014 Nobel Laureates in Chemistry for their achievements in super-resolution microscopy.
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