High contrast imaging and flexible photomanipulation for quantitative in vivo multiphoton imaging with polygon scanning microscope

Journal of Biophotonics
Yongxiao LiWoei Ming Lee

Abstract

In this study, we introduce two key improvements that overcome limitations of existing polygon scanning microscopes while maintaining high spatial and temporal imaging resolution over large field of view (FOV). First, we proposed a simple and straightforward means to control the scanning angle of the polygon mirror to carry out photomanipulation without resorting to high speed optical modulators. Second, we devised a flexible data sampling method directly leading to higher image contrast by over 2-fold and digital images with 100 megapixels (10 240 × 10 240) per frame at 0.25 Hz. This generates sub-diffraction limited pixels (60 nm per pixels over the FOV of 512 μm) which increases the degrees of freedom to extract signals computationally. The unique combined optical and digital control recorded fine fluorescence recovery after localized photobleaching (r ~10 μm) within fluorescent giant unilamellar vesicles and micro-vascular dynamics after laser-induced injury during thrombus formation in vivo. These new improvements expand the quantitative biological-imaging capacity of any polygon scanning microscope system.

References

Feb 11, 2000·Cell Calcium·N Callamaras, I Parker
Sep 27, 2002·Biophysical Journal·James D LechleiterIlse Sieneart
Jun 13, 2003·Biomedical Engineering Online·Thomas A PologrutoKarel Svoboda
May 28, 1965·Annals of the New York Academy of Sciences·J A KOCHEN, S BAEZ
May 17, 2005·Nature Neuroscience·Dimitrios DavalosWen-Biao Gan
Dec 3, 2005·The Journal of Clinical Investigation·Bruce Furie, Barbara C Furie
Mar 24, 2007·The Journal of Clinical Investigation·Christophe DuboisBruce Furie
Mar 7, 2005·Optics Express·Dan OronYaron Silberberg
Dec 19, 2009·Proceedings of the National Academy of Sciences of the United States of America·T DertingerJ Enderlein
Jul 1, 2010·Optics Express·Paul A KirkbyR Angus Silver
Dec 18, 2010·Optics Express·Hendrik DeschoutKevin Braeckmans
May 18, 2011·Proceedings of the National Academy of Sciences of the United States of America·Myunghwan ChoiChulhee Choi
Oct 25, 2011·Nature Biotechnology·Lacramioara IvanciuRodney M Camire
Sep 23, 2014·Nature Photonics·Guoan ZhengChanghuei Yang
Apr 4, 2015·Cold Spring Harbor Protocols·Heather A Sullivan, Ian R Wickersham
May 15, 2015·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Ben E UrbanHao F Zhang
Oct 30, 2015·Science Translational Medicine·Uma Mahesh R AvulaJérôme Kalifa
Jan 8, 2016·Scientific Reports·Mary Ann GoVincent Ricardo Daria
Jun 28, 2016·Nature Biotechnology·Jeffrey N StirmanSpencer L Smith
May 10, 2017·Nature Methods·Jung-Hoon ParkMeng Cui

❮ Previous
Next ❯

Citations

Feb 23, 2020·Frontiers in Immunology·Samantha J MontagueElizabeth E Gardiner

❮ Previous
Next ❯

Related Concepts

Related Feeds

Blood Clotting Disorders

Thrombophilia includes conditions with increased tendency for excessive blood clotting. Blood clotting occurs when the body has insufficient amounts of specialized proteins that make blood clot and stop bleeding. Here is the latest research on blood clotting disorders.