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Full text release has been delayed at the author's request until May 10, 2024

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High Resolution Fluorescence Imaging to Study Dynamics of Clathrin-Coated Structures

Akatay, Ahmet A
http://rave.ohiolink.edu/etdc/view?acc_num=osu1546634008760321

Abstract Details

2019, Doctor of Philosophy, Ohio State University, Biophysics.
Dynamics of clathrin-coated structures (CCS) using various fluorescent imaging modalities are studied. In order to image single molecule dynamics in real-time we developed an intense surface illumination technique called skewed illumination (SKILL). Super-resolved dynamics of CCS are reconstructed by real-time bleaching assisted localization microscopy (rtBALM) imaging under SKILL. It is found that exchange of clathrin coat components and remodeling of the CCS likely to occur at the peripheral regions flat CCS. Confocal 3D imaging showed that removals of coat components also localize to the edges of the structures. These results support that CCS dynamics are localized towards their perimeters. Analysis of 3D reconstructions of cell protrusions imaged under light-sheet illumination showed that a dense population of CCS, likely to be dominated by small clathrin-coated plaques, associates with the actin retrograde flow. Their dynamics suggest they exhibit adhesive properties. The dynamics of adhesive small clathrin-coated plaques and remodeling of flat CCS around their perimeters confirm their reported adhesive and mechanosensitive roles. A novel optical design methodology, called skewed optics, is introduced and implemented for surface illumination, i.e. SKILL. It is suggested to outperform the fast decaying illumination of total-internal reflection (TIR).
Richard Fishel (Advisor)
Comert Kural (Committee Member)
Ralf Bundschuh (Committee Member)
Carlos Castro (Committee Member)
192 p.
Biophysics; Optics

Recommended Citations

Citations

  • Akatay, A. A. (2019). High Resolution Fluorescence Imaging to Study Dynamics of Clathrin-Coated Structures [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1546634008760321

    APA Style (7th edition)

  • Akatay, Ahmet. High Resolution Fluorescence Imaging to Study Dynamics of Clathrin-Coated Structures . 2019. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1546634008760321.

    MLA Style (8th edition)

  • Akatay, Ahmet. "High Resolution Fluorescence Imaging to Study Dynamics of Clathrin-Coated Structures ." Doctoral dissertation, Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1546634008760321

    Chicago Manual of Style (17th edition)

osu1546634008760321
© 2019, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.