Open Collections

Zhang, Youchang

University of British Columbia

Among various super-resolution techniques, structured illumination microscopy (SIM) is the most suitable method for live-cell imaging owing to its high speed and low phototoxicity. However, the adoption of SIM faces challenges because of its technical limitations, such as reconstruction artifacts, reduced contrast from the missing cone problem, and the trade-off between spatial resolution and imaging speed. To circumvent these limitations, I present a new method to accomplish 2D SIM and 3D SIM imaging by employing a spinning disk. Spinning disk 2D SIM doubles the resolution in lateral directions, while spinning disk 3D SIM doubles the resolution both laterally and axially. They both achieve optical sectioning capability. In spinning disk SIM, the disk modulates the object frequency with a patterned illumination. By transmitting the patterned excitation through the disk, demodulation is accomplished, and super-resolved images with improved contrast are optically reconstructed in real-time. This thesis presents two disk designs which aim to maximize the resolution enhancement for 2D SIM and 3D SIM. The performance of the spinning disk SIM is examined though simulation and validated experimentally by imaging fluorescent beads and ryanodine receptors in a rat cardiac myocyte. Compared to wide-field microscopy which has a lateral resolution of approximately 250 nm and axial resolution around 800nm, the spinning disk 2D SIM provides a lateral resolution down to approximately 120 nm; the spinning disk 3D SIM reduced the axial resolution to around 400 nm.

Text

2024-09-30

Attribution-NonCommercial-NoDerivatives 4.0 International

http://hdl.handle.net/2429/85738

Chemistry

University of British Columbia

UBCV

http://creativecommons.org/licenses/by-nc-nd/4.0/