Jun. 2018

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

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Long-time two-photon imaging technology

doi:10.3788/CO.20181103.0337

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 ProgramNo.2017YFC0110303

National Major Scientific Research and Equipment Development ProjectNo.ZDYZ2013-1

Youth Foundation of Jiangsu Province Basic Research ProgramNo.BK20160363

Suzhou Applied Basic Research ProjectNo.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

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References (22)

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