长时程双光子成像技术

徐依雯; 张运海; 杨皓旻; 刘创; 唐玉国

doi:10.3788/CO.20181103.0337

Volume 11 Issue 3

Jun. 2018

Turn off MathJax

Article Contents

Chinese Optics > 2018 > 11(3): 337-343.

XU Yi-wen, ZHANG Yun-hai, YANG Hao-min, LIU Chuang, TANG Yu-guo. Long-time two-photon imaging technology[J]. Chinese Optics, 2018, 11(3): 337-343. doi: 10.3788/CO.20181103.0337

Citation:

XU Yi-wen, ZHANG Yun-hai, YANG Hao-min, LIU Chuang, TANG Yu-guo. Long-time two-photon imaging technology[J]. Chinese Optics, 2018, 11(3): 337-343. doi: 10.3788/CO.20181103.0337

XU Yi-wen, ZHANG Yun-hai, YANG Hao-min, LIU Chuang, TANG Yu-guo. Long-time two-photon imaging technology[J]. Chinese Optics, 2018, 11(3): 337-343. doi: 10.3788/CO.20181103.0337

Citation:

XU Yi-wen, ZHANG Yun-hai, YANG Hao-min, LIU Chuang, TANG Yu-guo. Long-time two-photon imaging technology[J]. Chinese Optics, 2018, 11(3): 337-343. doi: 10.3788/CO.20181103.0337

PDF( 2608 KB)

Long-time two-photon imaging technology

doi: 10.3788/CO.20181103.0337

XU Yi-wen^{1, 2
,},
ZHANG Yun-hai^{1
,
,},
YANG Hao-min¹,
LIU Chuang^{1, 2},
TANG Yu-guo¹

1.
Jiangsu Key Laboratory of Medical Optics, Suzhou Institute of Biomedical Engineering and Technology(SIBET), Chinese Academy of Sciences, Suzhou, Jiangsu 215163, China
2.
University of Chinese Academy of Sciences, Beijing 100049, China

Funds:

National Key R & D Program No.2017YFC0110303

National Major Scientific Research and Equipment Development Project No.ZDYZ2013-1

Youth Foundation of Jiangsu Province Basic Research Program No.BK20160363

Suzhou Applied Basic Research Project No.SYG201510

More Information

Corresponding author: ZHANG Yun-hai, E-mail:zhangyh@sibet.ac.cn
Received Date: 23 Jan 2018
Rev Recd Date: 13 Feb 2018
Publish Date: 01 Jun 2018

Abstract

Abstract

Two-photon imaging technology is widely used for dynamic 3D imaging of live cells due to its superior properties. However, severe photobleaching of fluorescence molecular on focal plane caused by short light pulse with extreme high optical power greatly affects long-term two-photon imaging. This paper proposes a method called Optimized Lighting -Two Photon(OL-TP)for the problem of two-photon fluorescence bleaching. With this method we obtain the high and low thresholds of the two-photon image through pre-scanning, optimize the duration of light in different areas of the image with preset high and low thresholds, and reconstruct the OL-TP image by using the fluorescence information and the illumination time information recorded during the scanning.Therefore, both the signal-to-noise ratio and fluorescence bleaching are guaranteed. The reconstructed OL-TP image is almost the same as the conventional two-photon image, though the signal-to-noise ratio is slightly reduced, the image is not distorted. For the 110 nm fluorescent ball samples, 30 normal two-photon images and optimized light two-photon images were successively taken. By the 30th image, the OL-TP imaging technology reduces 28.86% of photobleaching compared with standard two-photon images. All in all, OL-TP greatly reduces the photobleaching of two-photon imaging by optimizing the illumination time, enabling the two-photon fluorescence microscope to better observe biospecimen for long periods of time.
- two-photon microscopy imaging,
- fluorescence microscopy imaging,
- photobleaching

FullText(HTML)

References(22)

References

[1]	CRISAFI F, KUMAR V, PERRI A, et al.. Multimodal nonlinear microscope based on a compact fiber-format laser source[J]. Spectrochimica Acta Part A Molecular & Biomolecular Spectroscopy, 2017, 188:135-140. http://cn.bing.com/academic/profile?id=ef8ab5e58d2ad5ba69a3d119d127365c&encoded=0&v=paper_preview&mkt=zh-cn
[2]	DENK W, STRICKLER J H, WEBB W W. Two-photon laser scanning fluorescence microscopy[J]. Science, 1990, 248(4951):73-76. doi: 10.1126/science.2321027
[3]	李茜. 飞秒金宝搏188软件怎么用双光子荧光生物显微成像研究[D]. 哈尔滨: 哈尔滨工业大学, 2014. http://cdmd.cnki.com.cn/Article/CDMD-10213-1014083095.htm LI Q. Femtosecond laser two-photon fluorescence biological microscopy[D]. Harbin: Harbin Institute of Technology, 2014. (in Chinese) http://cdmd.cnki.com.cn/Article/CDMD-10213-1014083095.htm
[4]	李奇峰, 沙乾坤, 王洋, 等.基于非线性光学的2PE-STED显微技术的发展与应用[J].纳米技术与精密工程, 2015(2):81-89. http://www.cnki.com.cn/Article/CJFDTotal-NMJM201502001.htm LI Q F, SHA Q K, WANG Y, et al.. Development and application of 2PE-STED microscopy based on nonlinear optics[J]. Nanotechnology and Prescision Engineering, 2015(2):81-89.(in Chinese) http://www.cnki.com.cn/Article/CJFDTotal-NMJM201502001.htm
[5]	MILLER D R, JARRETT J W, HASSAN A M, et al.. Deep tissue imaging with multiphoton fluorescence microscopy[J]. Current Opinion in Biomedical Engineering, 2017, 4:32-39. doi: 10.1016/j.cobme.2017.09.004
[6]	TSAKANOVA G, ARAKELOVA E, AYVAZYAN V, et al.. Two-photon microscopy imaging of oxidative stress in human living erythrocytes[J]. Biomedical Optics Express, 2017, 8(12):5834-5846. doi: 10.1364/BOE.8.005834
[7]	熊大曦, 刘云, 梁永, 等.共振扫描显微成像中的图像畸变校正[J].光学精密工程, 2015, 23(10):2971-2979. http://www.eope.net/gxjmgc/article/2015/2015-10-2971.htm XIONG D X, LIU Y, LIANG Y, et al.. Correction of distortion in microscopic imaging with resonant scanning[J]. Opt. Precision Eng., 2015, 23(10):2971-2979.(in Chinese) http://www.eope.net/gxjmgc/article/2015/2015-10-2971.htm
[8]	马军, 田玉顺.双光子荧光影像技术在药物研发中的应用前景[J].中国现代应用药学, 2016, 33(3):381-385. http://www.cqvip.com/QK/95393A/201603/668383114.html MA J, TIAN Y SH. Application prospect of two-photon fluorescence imaging technology in drug development[J]. Chinese Journal of Modern Applied Pharmacy, 2016, 33(3):381-385.(in Chinese) http://www.cqvip.com/QK/95393A/201603/668383114.html
[9]	崔权, 陈忠云, 张智红, 等.多色双光子成像技术进展[J].金宝搏188软件怎么用与光电子学进展, 2017(6):10-23. http://www.opticsjournal.net/ViewFullPDF.htm?aid=OJ170608000009LiOkRn CUI Q, CHEN ZH Y, ZHANG ZH H, et al.. Recent advances in multicolor two-photon imaging technique[J]. Laser & Optoelectronics Progress, 2017, 54(6):060002.(in Chinese) http://www.opticsjournal.net/ViewFullPDF.htm?aid=OJ170608000009LiOkRn
[10]	LEFORT C. A review of biomedical multiphoton microscopy and its laser sources[J]. Journal of Physics D Applied Physics, 2017, 50(42):423001. doi: 10.1088/1361-6463/aa8050
[11]	TISCHBIREK C H, BIRKNER A, KONNERTH A. In vivo deep two-photon imaging of neural circuits with the fluorescent Ca²⁺ indicator Cal-590[J]. Journal of Physiology, 2017, 595(10):3097-3105. doi: 10.1113/JP272790
[12]	夏伟强, 周源, 石明.双光子显微成像技术的新进展[J].中国医疗器械杂志, 2011, 35(3):204-208. http://www.docin.com/p-304442607.html XIA W Q, ZHOU Y, SHI M. Advances in two-photon imaging technology[J]. Chinese Journal of Medical Instrumentation, 2011, 35(3):204-208.(in Chinese) http://www.docin.com/p-304442607.html
[13]	HOPT A, NEHER E. Highly nonlinear photodamage in two-photon fluorescence microscopy[J]. Biophys J, 2001, 80(4):2029-2036. doi: 10.1016/S0006-3495(01)76173-5
[14]	STEPHENS D J, ALLAN V J. Light microscopy techniques for live cell imaging[J]. Science, 2003, 300(5616):82-86. doi: 10.1126/science.1082160
[15]	STAUDT T, ENGLER A, RITTWEGER E, et al.. Far-field optical nanoscopy with reduced number of state transition cycles[J]. Optics Express, 2011, 19(6):5644-5657. doi: 10.1364/OE.19.005644
[16]	HOEBE R A, VAN OVEN C H, GADELLA T W J, et al.. Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence live-cell imaging[J]. Nat. Biotech., 2007, 25(2):249-253. doi: 10.1038/nbt1278
[17]	MONDAL P P, DIASPRO A. Reduction of higher-order photobleaching in two-photon excitation microscopy[J]. Physical Review E Statistical Nonlinear & Soft Matter Physics, 2007, 75(6 Pt 1):061904. http://cn.bing.com/academic/profile?id=f241922887607447309f38575d1a6513&encoded=0&v=paper_preview&mkt=zh-cn
[18]	PATTERSON, GEORGE H, DAVID W. Photobleaching in two-photon excitation microscopy[J]. Biophysical Journal, 2000, 78(4):2159-2162. doi: 10.1016/S0006-3495(00)76762-2
[19]	张运海, 杨皓旻, 孔晨晖.金宝搏188软件怎么用扫描共聚焦光谱成像系统[J].光学精密工程, 2014, 22(6):1446-1453. http://www.eope.net/gxjmgc/CN/abstract/abstract15267.shtml ZHANG Y H, YANG H M, KONG CH H. Spectral imaging system on laser scanning confocal microscopy[J]. Opt. Precision Eng., 2014, 22(6):1446-1453.(in Chinese) http://www.eope.net/gxjmgc/CN/abstract/abstract15267.shtml
[20]	EGGELING C, WILLIG K I, SAHL S J, et al.. Lens-based fluorescence nanoscopy[J]. Quarterly Reviews of Biophysics, 2015, 48(2):178-243. doi: 10.1017/S0033583514000146
[21]	张达奇. 全参考图像主观质量客观评价算法研究[D]. 苏州: 苏州大学, 2014. http://cdmd.cnki.com.cn/Article/CDMD-10285-1014298659.htm ZHANG D Q. Research on objective algorithms for full-reference image quality assessment[D]. Soochow: Soochow University, 2014. (in Chinese) http://cdmd.cnki.com.cn/Article/CDMD-10285-1014298659.htm
[22]	褚江, 陈强, 杨曦晨.全参考图像质量评价综述[J].计算机应用研究, 2014, 31(1):13-22. http://www.cnki.com.cn/Article/CJFDTotal-JSYJ201401004.htm CHU J, CHEN Q, YANG X CH. Review on full reference image quality assessment algorithms[J]. Application Research of Computers, 2014, 31(1):13-22.(in Chinese) http://www.cnki.com.cn/Article/CJFDTotal-JSYJ201401004.htm

Relative Articles

Supplements(0)

Cited By

Proportional views

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Figures(7)

Get Citation

PDF

XML

Article views(2603) PDF downloads(234)

Long-time two-photon imaging technology

doi: 10.3788/CO.20181103.0337

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Proportional views

Related

Long-time two-photon imaging technology

doi: 10.3788/CO.20181103.0337

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Proportional views

Related

Export File

Citation

Format

Content