Date of Award

Fall 1-1-2025

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biomedical Engineering (ENAS)

First Advisor

Bewersdorf, Joerg

Abstract

Advances in expansion microscopy, a super resolution microscopy technique in which samplesare expanded up to ~8,000 times their original volume, require new microscope modalities to image the large sample sizes. These new techniques should be suitable for imaging large sample volumes, yet need to maintain the high 3D resolution of 250-500 nm of high-end confocal microscopy to provide the maximum level of detail and thereby reveal nanoscale cellular ultrastructure. Axially Swept Light Sheet Microscopy (ASLM) has emerged as a promising technique for generating high-resolution images at high-throughput. However, current designs struggle with the trade-off between limited resolution and field-of-view (FOV) with slow image acquisition speeds. To enable high-throughput imaging of pan-Expansion Microscopy (pan-ExM) samples, I have designed and built a high-resolution, large-FOV light sheet microscope called pan-ASLM, that provides to our knowledge the fastest speed and highest FOV-to-resolution ratio among current high-resolution ASLM variants. pan-ASLM provides ~500 nm isotropic resolution over centimeter-scale image volumes at a high data acquisition speed of up to 20 fps (183 Megavoxels/sec). I validated the new microscope design by imaging pan-expanded HeLa cells, as well as mouse brain and kidney tissue. The new microscope has promise to unlock high-throughput nanoscale 3D imaging of expanded samples dramatically accelerating biological discoveries.

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