Name: implementation of a coherent anti-stokes raman scattering (cars) system on a ti:sapphire and opo laser based standard laser scanning microscope
Uploaded: 2016-07-17T14:00:00
Description: Watch this Scientific Journal Video about Implementation of a Coherent Anti-Stokes Raman Scattering CARS System on a Ti:Sapphire and OPO Laser Based Standard Laser Scanning Microscope at JoVE.com

The authors want to thank Dr. Philippe Combette (IES, UM, Montpellier, France) for the loan of the fast oscilloscope and acknowledge financial supports from Montpellier RIO Imaging (MRI). HR acknowledges ANR grants France Bio Imaging (ANR-10-INSB-04-01) and France Life Imaging (ANR-11-INSB-0006) infrastructure networks for coherent Raman imaging developments. This work was mainly supported by an European Research Council grant (FP7-IDEAS-ERC 311610) and an INSERM - AVENIR grant to NT.

<img alt="그림 1" src="/files/ftp_upload/54262/54262fig1.jpg" /> 그림 1. 일반적인 셋업의 개략도 그것은 티타늄과 같습니다. 사파이어 (680 - 1080 ㎚)와 OPO (1,050 - 1,300 ㎚) 레이저, (M 4 M 1) 4 거울 지연 라인을, 빠른 오실로스코프, 포토 다이오드와 두 개의 고정 조리개 진동판 I 1 I 2. 미러 M 2 및 M 3 마이크로 미터...

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	<li>Valeur, B., Berberan-Santos, M. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Molecular Fluorescence: Principles and Applications. , (2012).</li><li>Denk, W., Strickler, J. H., Webb, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-photon+laser+scanning+fluorescence+microscopy.">Two-photon laser scanning fluorescence microscopy.</a> Science. 248 (4951), 73-76 (1990).</li><li>Moreaux, L., Sandre, O., Mertz, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+imaging+by+second-harmonic+generation+microscopy.">Membrane imaging by second-harmonic generation microscopy.</a> JOSA B. 17 (10), 1685-1694 (2000).</li><li>Zoumi, A., Yeh, A., Tromberg, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+cells+and+extracellular+matrix+in+vivo+by+using+second-harmonic+generation+and+two-photon+excited+fluorescence.">Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence.</a> Proc. Natl. Acad. Sci. USA. 99 (17), 11014-11019 (2002).</li><li>Yelin, D., Silberberg, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+scanning+third-harmonic-generation+microscopy+in+biology.">Laser scanning third-harmonic-generation microscopy in biology.</a> Opt. Express. 5 (8), 169-175 (1999).</li><li>Campagnola, P. J., Millard, A. C., Terasaki, M., Hoppe, P. E., Malone, C. J., Mohler, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+high-resolution+Second-Harmonic+Generation+imaging+of+endogenous+structural+proteins+in+biological+tissues.">Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues.</a> Biophys. J. 81 (1), 493-508 (2002).</li><li>Olivier, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+lineage+reconstruction+of+early+zebrafish+embryos+using+label-free+nonlinear+microscopy.">Cell lineage reconstruction of early zebrafish embryos using label-free nonlinear microscopy.</a> Science. 329 (5994), 967-971 (2010).</li><li>Farrar, M. J., Wise, F. W., Fetcho, J. R., Schaffer, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+of+myelin+in+the+vertebrate+central+nervous+system+using+third+harmonic+generation+microscopy.">In vivo imaging of myelin in the vertebrate central nervous system using third harmonic generation microscopy.</a> Biophys. J. 100 (5), 1362-1371 (2011).</li><li>Lim, H., Sharoukhov, D., Kassim, L., Zhang, Y., Salzer, J. L., Melendez-Vasquez, C. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Label-free+imaging+of+Schwann+cell+myelination+by+third+harmonic+generation+microscopy.">Label-free imaging of Schwann cell myelination by third harmonic generation microscopy.</a> Proc. Natl. Acad. Sci. U.S.A. 111 (50), 18025-18030 (2014).</li><li>Strupler, M., Pena, A. M., Hernest, M., Tharaux, P. L., Martin, J. L., Beaurepaire, E., Schanne-Klein, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Second+harmonic+imaging+and+scoring+of+collagen+in+fibrotic+tissues.">Second harmonic imaging and scoring of collagen in fibrotic tissues.</a> Opt. Express. 15 (7), 4054-4065 (2007).</li><li>Cheng, J. X., Xie, X. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coherent+anti-Stokes+Raman+scattering+microscopy:+Instrumentation,+theory,+and+applications.">Coherent anti-Stokes Raman scattering microscopy: Instrumentation, theory, and applications.</a> J. Phys. Chem. B. 108 (3), 827-840 (2004).</li><li>Volkmer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vibrational+imaging+and+microspectroscopies+based+on+coherent+anti-Stokes+scattering+microscopy.">Vibrational imaging and microspectroscopies based on coherent anti-Stokes scattering microscopy.</a> J. Phys. D: Appl. Phys. 38, R59-R81 (2005).</li><li>Evans, C. L., Xie, X. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coherent+anti-Stokes+Raman+scattering+microscopy:+chemical+imaging+for+biology+and+medicine.">Coherent anti-Stokes Raman scattering microscopy: chemical imaging for biology and medicine.</a> Annu. Rev. Anal. Chem. 1, 883-909 (2008).</li><li>Mukamel, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Principles of nonlinear optical spectroscopy. , (1995).</li><li>Duncan, M. D., Reintjes, J., Manuccia, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scanning+coherent+anti-Stokes+Raman+microscope.">Scanning coherent anti-Stokes Raman microscope.</a> Opt. Lett. 7 (8), 350-352 (1982).</li><li>Zumbusch, A., Holtom, G. R., Xie, X. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+vibrational+imaging+by+coherent+anti-Stokes+Raman+scattering.">Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering.</a> Phys. Rev. Lett. 82 (20), 4142-4145 (1999).</li><li>Folick, A., Min, W., Wang, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Label-free+imaging+of+lipid+dynamics+using+Coherent+Anti-Stokes+Raman+Scattering+(CARS)+and+Stimulated+Raman+Scattering+(SRS)+microscopy.">Label-free imaging of lipid dynamics using Coherent Anti-Stokes Raman Scattering (CARS) and Stimulated Raman Scattering (SRS) microscopy.</a> Curr. Opin. Genet. Dev. 21 (5), 585-590 (2011).</li><li>Wang, P., Liu, B., Zhang, D., Belew, M. Y., Tissenbaum, H. A., Cheng, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+lipid+metabolism+in+live+Caenorhabditis+elegans+using+fingerprint+vibrations.">Imaging lipid metabolism in live Caenorhabditis elegans using fingerprint vibrations.</a> Angew. Chem. Int. Ed. Engl. 53 (44), 11787-11792 (2014).</li><li>Min, W., Freudiger, C. W., Lu, S., Xie, X. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coherent+nonlinear+optical+imaging:+beyond+fluorescence+microscopy.">Coherent nonlinear optical imaging: beyond fluorescence microscopy.</a> Annu. Rev. Phys. Chem. 62, 507-530 (2011).</li><li>Cheng, J. X., Jia, Y. K., Zheng, G., Xie, X. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser-scanning+coherent+anti-Stokes+Raman+scattering+microscopy+and+applications+to+cell+biology.">Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology.</a> Biophys J. 83 (1), 502-509 (2002).</li><li>Chiu, W. S., Belsey, N. A. N., Garrett, L., Moger, J., Delgado-Charro, M. B., Guy, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+diffusion+in+the+human+nail+measured+by+stimulated+Raman+scattering+microscopy.">Molecular diffusion in the human nail measured by stimulated Raman scattering microscopy.</a> Proc Natl. Acad. Sci. U.S.A. 112, 7725-7730 (2015).</li><li>Chen, X., Gr&#233;goire, S., Formanek, F., Galey, J. -. B., Rigneault, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+3D+molecular+cutaneous+absorption+in+human+skin+using+label+free+nonlinear+microscopy.">Quantitative 3D molecular cutaneous absorption in human skin using label free nonlinear microscopy.</a> J. of Control. Release. 200, 78-86 (2015).</li><li>Kano, H., Hamaguchi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+multi-nonlinear+optical+imaging+of+a+living+cell+using+a+supercontinuum+light+source+generated+from+a+photonic+crystal+fiber.">In vivo multi-nonlinear optical imaging of a living cell using a supercontinuum light source generated from a photonic crystal fiber.</a> Opt. Express. 14 (7), 2798-2804 (2006).</li><li>Brustlein, S., Ferrand, P., Walther, N., Brasselet, S., Billaudeau, C., Marguet, D., Rigneault, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+parametric+oscillator-based+light+source+for+coherent+Raman+scattering+microscopy:+practical+overview.">Optical parametric oscillator-based light source for coherent Raman scattering microscopy: practical overview.</a> J. Biomed. Opt. 16 (2), 021106 (2011).</li><li>Chen, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multimodal+platform+for+nonlinear+optical+microscopy+and+microspectroscopy.">A multimodal platform for nonlinear optical microscopy and microspectroscopy.</a> Opt. Express. 17 (3), 1282-1290 (2009).</li><li>Yue, S., Slipchenko, M. N., Cheng, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multimodal+nonlinear+optical+microscopy.">Multimodal nonlinear optical microscopy.</a> Laser Photonics Rev. 5 (4), 496-512 (2011).</li><li>Sun, Q., Li, Y., He, S., Situ, C., Wu, Z., Qu, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Label-free+multimodal+nonlinear+optical+microscopy+reveals+fundamental+insights+of+skeletal+muscle+development.">Label-free multimodal nonlinear optical microscopy reveals fundamental insights of skeletal muscle development.</a> Biomed Opt Express. 5 (1), 158-166 (2013).</li><li>Le, T. T., Langohr, I. M., Locker, M. J., Sturek, M., Cheng, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Label-free+molecular+imaging+of+atherosclerotic+lesions+using+multimodal+nonlinear+optical+microscopy.">Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy.</a> J. Biomed. Opt. 12 (5), 054007 (2007).</li><li>Ellis, D. I., Cowcher, D. P., Ashton, L., O'Hagana, S., Goodacre, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Illuminating+disease+and+enlightening+biomedicine:+Raman+spectroscopy+as+a+diagnostic+tool.">Illuminating disease and enlightening biomedicine: Raman spectroscopy as a diagnostic tool.</a> Analyst. 138, 3871-3884 (2013).</li><li>Oz&#231;elik, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pals1+is+a+major+regulator+of+the+epithelial-like+polarization+and+the+extension+of+the+myelin+sheath+in+peripheral+nerves.">Pals1 is a major regulator of the epithelial-like polarization and the extension of the myelin sheath in peripheral nerves.</a> J Neurosci. 30 (11), 4120-4131 (2010).</li><li>Heinrich, C., Hofer, A., Ritsch, A., Ciardi, C., Bernet, S., Ritsch-Marte, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+imaging+of+saturated+and+unsaturated+lipids+by+wide-field+CARS-microscopy.">Selective imaging of saturated and unsaturated lipids by wide-field CARS-microscopy.</a> Opt. Express. 16 (4), 2699-2708 (2008).</li><li>Kyriakidis, N. B., Skarkalis, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+spectra+measurement+of+olive+oil+and+other+vegetable+oils.">Fluorescence spectra measurement of olive oil and other vegetable oils.</a> J. AOAC Int. 83 (6), 1435-1439 (2000).</li><li>King, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscopic+anatomy:+normal+structure.">Microscopic anatomy: normal structure.</a> Handb. Clin. Neurol. 115, 7-27 (2013).</li><li>Monsma, P. C., Brown, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FluoroMyelin+Red+is+a+bright,+photostable+and+non-toxic+fluorescent+stain+for+live+imaging+of+myelin.">FluoroMyelin Red is a bright, photostable and non-toxic fluorescent stain for live imaging of myelin.</a> J. Neurosci. Methods. 209 (2), 344-350 (2012).</li><li>Wang, H., Fu, Y., Zickmund, P., Shi, R., Cheng, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coherent+anti-stokes+Raman+scattering+imaging+of+axonal+myelin+in+live+spinal+tissues.">Coherent anti-stokes Raman scattering imaging of axonal myelin in live spinal tissues.</a> Biophys. J. 89 (1), 581-591 (2005).</li><li>Wang, H. W., Fu, Y., Huff, T. B., Le, T. T., Wang, H., Cheng, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chasing+lipids+in+health+and+diseases+by+coherent+anti-Stokes+Raman+scattering+microscopy.">Chasing lipids in health and diseases by coherent anti-Stokes Raman scattering microscopy.</a> Vib. Spectrosc. 50 (1), 160-167 (2009).</li><li>Jung, Y., Tam, J., Jalian, H. R., Anderson, R. R., Evans, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal,+3D+in+vivo+imaging+of+sebaceous+glands+by+coherent+anti-stokes+Raman+scattering+microscopy:+normal+function+and+response+to+cryotherapy.">Longitudinal, 3D in vivo imaging of sebaceous glands by coherent anti-stokes Raman scattering microscopy: normal function and response to cryotherapy.</a> J. Invest. Dermatol. 135 (1), 39-44 (2015).</li></ol>

작품의 가장 어려운 부분은 레이저 빔의 시간 동기화된다. 그것은 빠른 오실로스코프와 결합 된 빠른 광 다이오드를 필요로하지만, 시간 만 거친 중복 처음에 수행 할 수 있습니다. 그 후 몇 cm의 추가적인 조정이 필요하다. 마지막으로, 선형 변환 스테이지 마이크로 미터에 의해 이동은 CARS 신호를 트리거하기 위해 지연 라인 길이의 최종 미세 조정을 행하는 허용한다. 변환 스테이지 마이크로 드?...

광학 현미경은 세포 내 해상도로 생물학적 시스템을 살고 동적 프로세스의 비파괴 시각화를위한 주요 기술이되었다. 형광 현미경으로 인해 높은 특이 도와 민감도 1 생균 현재 가장 많이 사용되는 영상의 대비이다. 형광 프로브의 큰 팔레트 (외인성 색소, 유전자 인코딩 단백질, 반도체 나노 입자를) 등장했습니다. 다양한 샘플 조명 형광 기반 기술은 3 차원 영상을 수행하고 2 photobleaching에되는이 기술의 주요 단점을 줄이기 위해 (예 촛점 또는 이광자 현미경 등) 번성하고있다. 분자 종의 대부분은 본질적으로 형광하지 않으며, 따라서 이러한 형광 인위적으로 몇 군데 샘플에 도입 할 필요가 있기 때문에 다른 제한은 형광 라벨의 요구 사항을 포함한다. 이 인공 조작은 작은 분자 특히 파괴적이거나 냄비를 유도 할 수있다ential 사진 독성. 이러한 이유는 생체 관측 형광 현미경하지가 적합합니다. 따라서, 형광성 분자의 사용없이 고감도 특정 분자 대조 광학 이미징 기술의 사용은 바이오 메디컬 사이언스에서 매우 바람직하다. 라벨이나 얼룩없이 여러 비선형 광학 이미징 기술은 두 번째 고조파 세대 (SHG) 3,4와 세 번째 고조파 세대 (THG) 5를 포함 등장했다. SHG 현미경은 미세 소관 또는 콜라겐 6으로 초분자 레벨에서 화상 구조 배치로 사용되어왔다. THG는 수성 매질 및 지질 (7) 사이의 인터페이스로서 광 이질성로부터 생성된다. THG는 이미지 수초 8,9로 증명되었다. 두 기술들은 이광자 형광 현미경상에서 구현 오직 하나의 레이저 빔을 필요로 할 수있다. 이들은 높은 전력 레이저 강도를 필요로하지만 (전형적으로 50생활 샘플에 해로운이며, 명확하게 이미지 특정 생물학적 구조에 필요한 화학적 특이성을 제공하지 않는 THG 9 1,180 ㎚), 50 mW의 - SHG (10), (25) 860 nm에서 mW의. 선택적 화학적 이미징 라만 산란에 기초한 분자 고유 진동 신호로부터 얻을 수있다. 광 빔 상관 닿으 광자 흡수 자나 분자에 의해 산란 될 수있다. 산란 된 광자의 대부분은 입사 광자 에너지와 같은, 즉, 주파수를 가질 것이다. 이 프로세스는 레일리 산란이라고한다. 그러나, 광자 소수 라만 산란이라 비탄성 산란 과정, 즉, 입사 광자의 주파수와 다른 주파수의 광 산란한다. 에너지의 차이는 분자 구조 및 환경에 따라 진동 모드의 여기로부터 기인한다. 따라서, 자발 라만 산란 잠십오 화학적으로 선택적 영상은 다른 분자는 특정 진동 주파수를 가지고있다. 그러나 그것 때문에 매우 약한 신호의 제한됩니다. 공 초점 라만 현미경이 개발 및 3D 공간 해상도를 제공하지만, 높은 평균 전력 및 긴 획득 시간 (11)을 필요로하고있다. 이러한 어려움을 극복하기 위해 레이저 기술의 최근 발전은 특히 코 히어 런트 반 스톡스 라만 산란의 비선형 광학 진동 현미경의 상승 (CARS) 11,12,13 수있다. CARS는 3 차 비선형 광학 과정이다. 주파수 ω의 P에 펌프광 구성된 세 개의 레이저 빔은 주파수 ω의 S에서 스톡스 광 및 프로브 빔 (주로 펌프 임) 시료에 집중되고 (주파수 ω AS =에서 반 스톡스 빔을 생성 2ω P - ω의 S) 14. 안티 - 스토크 스 신호를 크게 향상시킬 수있을 때, 주파수 차이(- ω S ω P) 펌프와 스톡스 라만 분자 진동 Ω의 R =로 조정되어 빔 사이. CARS 신호는 다수의 광자 상호 작용을 기반으로합니다. 따라서, 자발 라만 산란 강도보다 강한 간섭 신호의 순서를 생성한다. CARS 현미경은 제 실험적 던컨 등. (15)에 의해 입증되었다. Zumbusch 외는. 높은 개구 수의 대물 렌즈를 두 초점 근적외선 펨토초 레이저 빔을 이용하여 CARS의 위상 정합 조건을 허용하고 이광자 비 공진 배경 (16)을 회피하여, 그 기술을 향상시켰다. CARS 현미경 따라서 화학적으로 살아있는 세포 (19, 20)을로 (CH 신축 진동을 통해) 지질 같은 분자 (17, 18), 물 (OH 신축 진동을 통해), 단백질, DNA를 검출함으로써, 살아있는 세포와 조직 이미징을위한 강력한 도구로 떠오르고있다 또한 화학 물질 화합물을 중수 소화제약 (21) 및 화장품 응용 프로그램 (22)에 대한의. 비선형 현미경의 가장 큰 한계는 복잡성과 광원의 비용에서 기인한다. CARS 시스템은 짧은 펄스 기간과 시간적 및 공간적 동기화 펄스 트레인 두 파장 가변 레이저가 필요합니다. 초기 CARS 현미경은 두 개의 동기화 된 피코 초 티를 기반으로했다 : 사파이어 레이저 (20). 초 연속 광원 (23)을 생성하는 사파이어 레이저 : CARS 이미징은 단일 펨토초 TI의 얻었다. 최근에, 단일 펨토초 Ti로 이루어지는 레이저 소스 : 가변 광 파라 메트릭 발진기 (OPO)를 펌핑 사파이어 레이저는 CARS 현미경에 사용되어왔다. 이 셋업은 본질적으로 일시적으로 펌프와 전체 분자 진동 스펙트럼 (24)을 포함 스톡스 빔 사이의 주파수의 차이 빔을 동기화 허용한다. 또한, 레이저 주사 현미경 턴에 기초주로 두 광자 형광 (TPF)에 사용되는 키 FS 레이저와 OPO가 아닌 물리학에 대해 사용할 수 있습니다. 각 비선형 (NLO) 영상 기법은 특정 구조 나 분자에 민감하기 때문에 이러한 셋업의 전위가 크게 다른 비선형 광학 영상의 혼입에 의해 보충 투​​자를 필요로하지 않고 향상 될 수있다. 복합 NLO 촬상 따라서 복잡한 생물학적 시료 25 NLO 현미경의 전위 대문자. 이러한 기술의 결합은 지질 대사, 피부 암 개발 (26), 골격 근육 발달 (27), 동맥 경화성 병변 (28)에 특히 많은 생물 학적 질문의 조사를 허용했다. 또한 CARS과 레이저 빔 주사의 구현은 고속 촬상, 즉 생체 역학 과정을 연구하기위한 매력적인 도구의 기능을 제공한다. 이 연구의 목적은 t을 구현하는 각 단계를 보여주는 것입니다표준 광자 레이저 스캐닝 현미경에 그는 CARS 기술. 현미경은 FSEC 티를 기반으로 : 사파이어 레이저와 OPO : 생물 학자를위한 소프트웨어에 의해 동작합니다 (티 의해 펌핑 레이저 사파이어). 통합 시간에 두 개의 빔을 동기화하기 위해 상기 레이저 빔 경로 중 하나의 길이를 조정함으로써 수행 하였다. 우리는 실험 광학의 기본적인 배경을 필요로이 기술의 단계별 구현을 설명합니다. 우리는 또한 CARS는 쥐의 좌골 신경의 수초에서 얻은 영상 설명, 우리는이 영상은 표준 두 광자 형광 기술과 두 번째 고조파 세대로, 다른 비선형 광학 영상을 동시에 수행 할 수 있습니다 보여줍니다.

<table border="0" cellpadding="0" cellspacing="0" width="1157">
	<tbody>
		<tr height="23">
			<td height="23" style="height:23px;width:419px;">Oscilloscope</td>
			<td style="width:120px;">Tektronix</td>
			<td style="width:317px;">TDS 520D&#160;</td>
			<td style="width:301px;">500 MHz&#160;</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:419px;">Photodetector</td>
			<td style="width:120px;">Thorlabs</td>
			<td>DET08C/M, T4290</td>
			<td>5 GHz InGaAs, 800-1700 nm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Ti:Sapphire laser Chameleon Ultra Family II</td>
			<td style="width:120px;">Coherent</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Optical parametric oscillator OPO Compact Family</td>
			<td style="width:120px;">APE Berlin</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Axio Examiner microscope LSM 7 MP</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Motorized periscope</td>
			<td style="width:120px;">Newport</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:419px;">Objective W Plan-Apochromat 20x/1.0</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Beam combiner</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Acousto-optic modulator</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">OPO power attenuator</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Photomultiplier tube</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">ZEN software</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td style="width:317px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Bandpass filters</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td>LSM BiG 1935-176</td>
			<td>400-480 nm ; 500-550 nm ; 465-610 nm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Dichroic mirror</td>
			<td style="width:120px;">Carl Zeiss</td>
			<td></td>
			<td>Cutoff wavelength 760 nm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Silver mirrors</td>
			<td style="width:120px;">Newport</td>
			<td>10D20ER.2&#160;</td>
			<td>&#955;/10, 480-20,000 nm , Quantity 4</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:419px;">Single-axis translation stage with standard micrometer</td>
			<td style="width:120px;">Thorlabs</td>
			<td style="width:317px;">PT1/M</td>
			<td>Quantity 1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Aluminium breadboard</td>
			<td style="width:120px;">Thorlabs</td>
			<td>MB1015/M&#160;</td>
			<td>Quantity 1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Mirror mount</td>
			<td style="width:120px;">Thorlabs</td>
			<td>KMSS/M&#160;</td>
			<td>Quantity 4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mirror holder for &#216;1&#34; Optics&#160;</td>
			<td style="width:120px;">Thorlabs</td>
			<td>MH25</td>
			<td>Quantity 4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Iris diaphragms&#160;</td>
			<td style="width:120px;">Thorlabs</td>
			<td>ID8/M</td>
			<td>Quantity 3</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Protective box</td>
			<td style="width:120px;">Thorlabs</td>
			<td>TB4, XE25L900/M, T205-1.0, RM1S</td>
			<td>Quantity 1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Optical posts</td>
			<td style="width:120px;">Thorlabs</td>
			<td>TR40/M, PH50/M, PH75/M, BA2/M</td>
			<td>Quantity 8 (lengths depending on the set-up)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">661-690 nm bandpass filter</td>
			<td>Semrock</td>
			<td>676/29 nm BrightLine&#174; single-band bandpass filter</td>
			<td>Quantity 1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Fluorescent beads</td>
			<td style="width:120px;">ThermoFisher</td>
			<td>TetraSpeck&#8482; Fluorescent Microspheres Size Kit</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Laser viewing card</td>
			<td style="width:120px;">Thorlabs</td>
			<td style="width:317px;"></td>
			<td>IR laser viewing card</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">Laser safety glass</td>
			<td style="width:120px;">Newport</td>
			<td style="width:317px;">LV-F22.P5L07&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:419px;">FluoroMyelin&#8482; Red Fluorescent Myelin Stain&#160;</td>
			<td style="width:120px;">ThermoFisher</td>
			<td>F34652</td>
			<td></td>
		</tr>
	</tbody>
</table>

implementation of a coherent anti-stokes raman scattering (cars) system on a ti:sapphire and opo laser based standard laser scanning microscope

펨토초 티타늄 합성 레이저 스캐닝 현미경 : 레이저 라인을 복제하는 사파이어 레이저 및 광 파라 메트릭 발진 (OPO)은 생물학 가능하게되었다. 이 시스템은 기본적으로 멀티 채널 이광자 형광 현미경을 위해 설계된다. 그러나, 임의의 수정없이, 상기 제 2 고조파 발생 (SHG) 또는 3 고조파 발생 (THG) 보완 비선형 광학 현미경은 구성 분자 또는 수성 중간의 라벨없는 결상을 가능하게 세트해서 수행 될 수있다 지질 인터페이스. 이러한 기술은 생체의 관찰에 적합하지만, 화학적 특성으로 제한된다. 선택적 화학적 이미징 라만 산란에 기초한 고유 진동 신호로부터 얻을 수있다. 공 초점 라만 현미경은 3D 공간 해상도를 제공하지만, 높은 평균 전력 및 긴 획득 시간이 필요하다. 이러한 어려움을 극복하기 위해, 레이저 기술의 최근 발전은 (STABLE)을 허용했다비선형 광학 진동 현미경의 opment, 특히 코 히어 런트 반 스톡스 라만에 (CARS)을 산란. CARS 현미경 따라서 화학적으로 매핑 (OH 신축 진동을 통해) (CH 신축 진동을 통해) 지질, 물, 단백질이나 DNA에 의해 생물학적 라이브 세포 이미징을위한 강력한 도구로 떠오르고있다. 본 연구에서 우리는 표준 OPO 결합 광자 레이저 스캐닝 현미경에 CARS 기술의 구현을 설명한다. 이것은 레이저 빔의 경로 중 하나의 길이를 조정함으로써 2 개의 레이저 라인의 시간 동기에 근거한다. 우리는 기존 광자 시스템에서이 기법의 단계별 구현을 제시한다. 실험 광학의 기본 배경은 도움이되고 제시된 시스템은 고가의 추가 장비가 필요하지 않습니다. 우리는 또한 쥐의 좌골 신경의 수초에 얻은 이미징 CARS을 설명하고, 우리는이 영상이 같은 표준 t 다른 비선형 광학 이미징, 동시에 수행 할 수 있음을 보여WO 광자 형광 기법 및 제 고조파 발생.

<html> 표준의 Ti 펄스열 주파수 : 사파이어 레이저는 일반적으로 약 80 메가 헤르츠이다. 레이저 사파이어 다음 OPO은 상기 티타늄으로 펌핑되기 때문에, 동일한 주파수를 갖는다. 최소 200 MHz의 빠른 오실로스코프 따라서 필요합니다. 범위의 빠른 포토 다이오드 (600)에 1,100 내지도 필요합니다. 사파이어와 OPO 신호 1 / (2 × 80 × 106) = 6.2 나노초의 시프트 : TI의이 때 최대 ...

Watch this Scientific Journal Video about Implementation of a Coherent Anti-Stokes Raman Scattering CARS System on a Ti:Sapphire and OPO Laser Based Standard Laser Scanning Microscope at JoVE.com

Implementation of a Coherent Anti-Stokes Raman Scattering CARS System on a Ti:Sapphire and OPO Laser Based Standard Laser Scanning Microscope

분자 결합의 고유 진동에 따라 코 히어 런트 반 스톡스 라만 산란 (CARS) 현미경 레이블없는 화학적으로 선택적 라이브 세포 이미징을 허용합니다. 사파이어 레이저와 OPO 레이저 본 작업은 펨토초의 Ti에 기초한 표준 광자 레이저 스캐닝 현미경에 상보 현미경 기술의 구현을 제공한다.

사파이어와 OPO 레이저 기반 표준 레이저 스캐닝 현미경 :에 Ti에 코 히어 런트 반 스톡스 라만 산란 (CARS) 시스템의 구현

Laser scanning microscopes combining a femtosecond Ti:sapphire laser and an optical parametric oscillator (OPO) to duplicate the laser line have become ...

The overall goal of this procedure is to implement a complimentary microscopy technique called CARS, which allows label-free, chemically selective, lifestyle imaging on a standard multi-photon laser scanning microscope based on a femto-second titanium-sapphire laser and an optical parameter oscillator. This method can help answer key questions in the field of biology or neuroscience such as understanding of lipid-related diseases, skin penetration mechanisms, or skin disorders. The main advantage of this technique is that it's easy to implement on a standard laser scanning microscopy based on Ti:Sapphire and OPO lasers.Demonstrating the procedure will be Hassan Boukhaddaoui, research associate and head of the imaging platform of the laboratory and Vasyl Mytskaniuk. Turn on the titanium-sapphire and OPO lasers as presented in the text protocol. Switch on the microscope computer.Turn on the microscope component switches. Start the software by double-clicking the icon on the desktop and set it up according to the text protocol. Then, place a dichroic mirror with a cutoff wavelength at 760 nanometers in the side port slider.Locate it in the infinity space above the objective nose piece. Set the narrow band pass filter and the reflector cube in front of PMT1 to record only the CARS signal at 670 nanometers to reproduce the presented results. Then, place specific narrow band filters in front of PMT3 for fluorescence observation of the myelin, and in front of PMT4 for SHG observation.In the software, set the signal on the detector with an ad hoc band pass filter. Open the light path tool in the setup manager menu in the acquisition tab, then activate the desired PMT and select a color for the channel. For example, use green for CARS, and magenta for SHG.The two beams that originate from the same titanium sapphire laser are not in sync when they reach the microscope because the OPO beam is delayed when it is generated. Follow this procedure to resynchronize them by adjusting the length of one of the laser beam pads by incorporating a delay line. The use of a fast photo diode and a fast oscilloscope is required.First, using BNC cables, connect the input channel CH1 of the oscilloscope to the electrical BNC laser output sync out. Next, connect the input channel CH2 to the photo diode. Then, choose the CH1 channel as the trigger by pressing trigger menu, then the main menu button source and then the side menu button that corresponds to the channel selected as CH1.Now position and fix with optical mounting posts to the photo diode in the focal plane of a 10x air microscope objective. In the Channels tool, define the titanium-sapphire laser wavelength at 830 nanometers at low power, which is less than 1%of the full power. In the Acquisition Mode tool, reduce the scan area to one point in order to illuminate the photo diode with the tiniest beam.Then switch on the laser scan by clicking on the continuous button. Press Autoset on the oscilloscope and manually move the photo diode to get a pulse train on the oscilloscope screen. Press the run/stop button to freeze the display.Next, switch off the titanium-sapphire laser scan. In the software, click the Channels tool, deselect the 830 nanometer laser and set the OPO signal to 1107 nanometers and low power. Then switch on the OPO laser scan, record the pulse trains of the OPO laser on the oscilloscope and switch off the OPO laser scan.Now, compare the temporal shift between the titanium-sapphire and the OPO signals. Calculate the length of the delay line, which will be used to position the mirrors. Before proceeding, put on safety goggles and remove any chain bracelets or watches from wrists.Now, open the laser titanium-sapphire line where the delay line will be implemented by removing the protective tubes. Then select a wavelength in the visible range in order to be able to easily observe the laser beam, for example, 700 nanometers at low power, and switch on the laser scan. Now put two iris diaphragms into the open laser line using the optical mounting posts.Position one iris at the exit of the delay line and place the other iris at the entrance of the periscope. Next, decrease the iris diaphragm aperture and adjust the diaphragm positions to fit the laser beam path then fix them on the optical table. Finally, adjust the vertical position of a third mobile iris diaphragm.The size diaphragms will serve as a control for the realignment procedure to check the position of the laser beam while successively positioning the four mirrors of the delay line. Now, place the mirror M1 on a compact kinematic mirror mount at the entrance of the delay line, and adjust its position and its orientation to maintain the beam height with the use of the mobile iris diaphragm. Then place mirrors M2 and M3 at 90 degrees onto the translation stage according to the calculated delay line length and adjust their orientation using the mobile iris diaphragm.Set M4 at the exit f the delay line just before the iris and carefully adjust its position and angle to fit the laser beam path through the two fixed iris diaphragms. Now position the laser viewing card at the output of the microscope objective and check the laser beam profile. If necessary, slightly adjust the orientation of M4 to observe a uniform bright disc.Lastly, reposition the fast photo diode under the laser beam in the sample focus plane. Then observe the temporal shift between the titanium-sapphire laser beam and the OPO beam on the oscilloscope. If necessary, change the delay line length by moving the whole translation stage to synchronize the pulses.The spacial overlap of the two beams needed for a CARS signal is obtained by visualizing fluorescent polystyrene beads fixed to a microscope slide. Focus on the beads using a 20x water objective. Now, in the Channels tool of the acquisition tab, add track one, or use the existing track.Select the wavelength at 830 nanometers and low power for the titanium-sapphire laser beam. Toggle the color to green in the track one box from the Channels window, and the PMT3 or the PMT4 box from the light path window. Next, add track two for the OPO laser beam and set the wavelength at 1107 nanometers and low power.Toggle the color to red in the track two box from the Channels window and in the PMT3 or the PMT4 box from the light path window. Now, select the maximum scan area and adjust the gain of both tracks to 600. Then sequentially apply the scan of the two beams onto the sample by clicking on continuous.Observe the image in the screen area in the 2D view. Increase the power of both lasers. If necessary, slightly move the focusing drive to find the focus plane of the beads.Finally, adjust the crop and zoom the image in on a single bead or on a group of adjacent beads. Next, use the periscope controller to overlap the beams in the XY plane. In the software, open the Maintain tab.Click on the system options and display the motorized periscope tool window. Further details are discussed in the text protocol. After overlapping images from both laser beams, synchronization is achieved.To achieve the precise temporal adjustment to activate CARS, prepare a glass slide with a droplet of olive oil and cover slip. Then using a 20x water immersion objective, focus on the edge of the cover slip. Now, on track one, set the wavelength to 830 nanometrs for the titanium-sapphire laser beam and at 1107 nanometers for the OPO.Toggle both lasers in track one to get a simultaneous scan. Start with both lasers at low power. Next, on the light path window, uncheck PMT4 and select PMT1.Then choose the maximum scan area and switch on the laser scans by clicking on the continuous button. Adjust the gain to 600 and, if necessary, increase the power to both lasers. Next, adjust the display intensity in the display view option control block.Then slowly move the translation stage of the delay line until the signal becomes significantly enhanced. Next, decrease the power to both lasers. Now, check whether the signal is a CARS signal by alternatively switching off one of the two lasers.If the signal intensity becomes weaker or disappears, a CARS signal has been obtained. Since the final system is dedicated to non-physicists, enclose the light path of the delay line with an enclosure box and tubes to avoid direct access to a harmful, nonvisible, high peak powered laser beam. However, do provide access to the translation stage knob.A fluro-myelin red dye, which has selectivity for myelin, was used to observe the myelin sheath. Simultaneous illumination at 830 and at 1095 nanometers provides a CARS signal. The circles correspond to the myelin sheath surround axons in a transversal cut.The same structure is found while overlapping CARS and fluorescence images. The level of detail of CARS and fluorescent labeling is very similar. Potentially obviating the need for dye use.The system can also simultaneously generate a second harmonic signal from the collagen fibers at the outer service of the sciatic nerves. A different detector is used to record a signal at 550 nanometers since a narrow band pass filter at 670 nanometers is used for the CARS signal recording. The fibers are illustrated by a false magenta color.The CARS signal alone clearly shows the myelin sheaths. By simultaneously visualizing both signals, the myelin sheaths in green are seen surrounded by collagen fibers in magenta. A CARS signal requires less energy than SHG or THG signals.CARS signals were achieved with four milliWatts at 830 nanometers and 13 milliWatts at 1095 nanometers, while 50 milliWatts at 1095 nanometers was required to obtain an SHG signal. After watching this video, you should have a good understanding how to modify step by step one of the laser beam paths to temporally synchronize both laser beams. The implementation of the delay line can be achieved in a few weeks by biologists with basic background in experimental optics or in collaboration with physicists.Don't forget that working with high power, invisible laser beams can be extremely hazardous. And precautions, like wearing goggles while performing the procedure and final enclosure of the laser beam path should be taken.

Research

JoVE Journal

Biology

Implementation of a Coherent Anti-Stokes Raman Scattering (CARS) System on a Ti:Sapphire and OPO Laser Based Standard Laser Scanning Microscope

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