Seeing in the Light: Using Expansion Microscopy to Achieve Super-Resolution in Transmitted Light

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Light microscopy is inherently limited in resolution by properties of light such as diffraction and interference to 170-250 nm. Expansion microscopy is a quickly-developing method which achieves super-resolution by using a swellable hydrogel to physically expand biological samples themselves, rather than depending on the properties of fluorophores. This thesis demonstrates that expansion microscopy is a feasible means for achieving super-resolution in transmitted light microscopy modes. Though it has only been used for fluorescence imaging in the past, here I show that samples prepared for expansion microscopy—including liver tissue slices and myofibrillar bundles—are observable using transmitted light. While the majority of … continued below

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Migliore, Julia R. December 2022.

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  • Migliore, Julia R.

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Light microscopy is inherently limited in resolution by properties of light such as diffraction and interference to 170-250 nm. Expansion microscopy is a quickly-developing method which achieves super-resolution by using a swellable hydrogel to physically expand biological samples themselves, rather than depending on the properties of fluorophores. This thesis demonstrates that expansion microscopy is a feasible means for achieving super-resolution in transmitted light microscopy modes. Though it has only been used for fluorescence imaging in the past, here I show that samples prepared for expansion microscopy—including liver tissue slices and myofibrillar bundles—are observable using transmitted light. While the majority of the original sample material is removed in the expansion process, the hydrogel retains visible evidence of these samples. These demonstrate increased detail under brightfield microscopy that is useful for characterization. Sarcomeric regions are identifiable by this method and are confirmed by fluorescence imaging. Thus, expansion microscopy is a means to bring super-resolution to transmitted light imaging and is entirely compatible with fluorescence for the localization of proteins of interest.

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  • December 2022

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  • Feb. 9, 2023, 4:50 p.m.

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  • Feb. 13, 2023, 8:52 a.m.

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Thumbnail image of item number 1 in: 'Seeing in the Light: Using Expansion Microscopy to Achieve Super-Resolution in Transmitted Light'.
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Migliore, Julia R. Seeing in the Light: Using Expansion Microscopy to Achieve Super-Resolution in Transmitted Light, thesis, December 2022; Denton, Texas. (https://digital.library.unt.edu/ark:/67531/metadc2048604/: accessed April 28, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; .

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