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Imaging effectiveness calculator for non-design microscope samples

Applied Optics

Anthony, Stephen M.; Miller, Philip R.; Timlin, Jerilyn A.; Polsky, Ronen

When attempting to integrate single-molecule fluorescence microscopy with microfabricated devices such as microfluidic channels, fabrication constraints may prevent using traditional coverslips. Instead, the fabricated devices may require imaging through material with a different thickness or index of refraction. Altering either can easily reduce the quality of the image formation (measured by the Strehl ratio) by a factor of 2 or more, reducing the signal-to-noise ratio accordingly. In such cases, successful detection of single-molecule fluorescence may prove difficult or impossible. Here we provide software to calculate the effect of non-design materials upon the Strehl ratio or ensquared energy and explore the impact of common materials used in microfabrication.

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TYPE Journal Article YEAR 2019
DOIOSTIScopus

Posters for AA/CE Reception

Kuether, Robert J.; Allensworth, Brooke M.; Backer, Adam; Chen, Elton Y.; Dingreville, Remi; Forrest, Eric C.; Knepper, Robert A.; Tappan, Alexander S.; Marquez, Michael P.; Vasiliauskas, Jonathan G.; Rupper, Stephen; Grant, Michael J.; Atencio, Lauren C.; Hipple, Tyler; Maes, Danae; Timlin, Jerilyn A.; Ma, Tian J.; Garcia, Rudy J.; Danford, Forest L.; Patrizi, Laura P.; Galasso, Jennifer; Draelos, Timothy J.; Gunda, Thushara; Venezuela, Otoniel; Brooks, Wesley A.; Anthony, Stephen M.; Carson, Bryan; Reeves, Michael; Roach, Matthew; Maines, Erin; Lavin, Judith M.; Whetten, Shaun R.; Swiler, Laura P.

Abstract not provided.

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TYPE Presentation YEAR 2018
OSTI

Delivering crispr: A review of the challenges and approaches

Drug Delivery

Lino, Christopher A.; Harper, Jason C.; Carney, James; Timlin, Jerilyn A.

Gene therapy has long held promise to correct a variety of human diseases and defects. Discovery of the Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR), the mechanism of the CRISPRbased prokaryotic adaptive immune system (CRISPR-associated system, Cas), and its repurposing into a potent gene editing tool has revolutionized the field of molecular biology and generated excitement for new and improved gene therapies. Additionally, the simplicity and flexibility of the CRISPR/Cas9 site-specific nuclease system has led to its widespread use in many biological research areas including development of model cell lines, discovering mechanisms of disease, identifying disease targets, development of transgene animals and plants, and transcriptional modulation. In this review, we present the brief history and basic mechanisms of the CRISPR/Cas9 system and its predecessors (ZFNs and TALENs), lessons learned from past human gene therapy efforts, and recent modifications of CRISPR/ Cas9 to provide functions beyond gene editing. We introduce several factors that influence CRISPR/ Cas9 efficacy which must be addressed before effective in vivo human gene therapy can be realized. The focus then turns to the most difficult barrier to potential in vivo use of CRISPR/Cas9, delivery. We detail the various cargos and delivery vehicles reported for CRISPR/Cas9, including physical delivery methods (e.g. microinjection; electroporation), viral delivery methods (e.g. adeno-associated virus (AAV); full-sized adenovirus and lentivirus), and non-viral delivery methods (e.g. liposomes; polyplexes; gold particles), and discuss their relative merits. We also examine several technologies that, while not currently reported for CRISPR/Cas9 delivery, appear to have promise in this field. The therapeutic potential of CRISPR/Cas9 is vast and will only increase as the technology and its delivery improves.

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TYPE Journal Article YEAR 2018
DOIOSTIScopus
Results 26–50 of 277
Results 26–50 of 277