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Optogenetics presents the ability to control membrane dynamics through the usage of transfected proteins (opsins) and light stimulation. However, as the field continues to grow, the original biological and stimulation tools used have become dated or limited in their uses. The usage of Organic Light Emitting Diodes (OLEDs) in optical

Optogenetics presents the ability to control membrane dynamics through the usage of transfected proteins (opsins) and light stimulation. However, as the field continues to grow, the original biological and stimulation tools used have become dated or limited in their uses. The usage of Organic Light Emitting Diodes (OLEDs) in optical stimulation offers greater resolution, finer control of pixel arrays, and the increased functionality of a flexible display at the cost of lower irradiance power density. This study was done to simulate methods using genetic and optical tools towards decreasing the threshold irradiance needed to initiate an action potential in a ChR2 expressing neuron. Simulations show that pulsatile stimulation can decrease threshold irradiances by increasing the overall duration of stimulus while keeping individual pulse durations below 5 ms. Furthermore, the redistribution of Channelrhodopsin-2 (ChR2) to the apical dendrites and a change in wavelength to 625 nm both result in lower threshold irradiances. However, the model used has many discrepancies and has room for improvement in areas such as the light distribution model and ChR2 dynamics. The simulations run with this model however still present valuable insight and knowledge towards the usage of new stimulation methods and revisions on existing protocols.
ContributorsKyeh, James (Author) / Muthuswamy, Jitendran (Thesis director) / Crook, Sharon (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description

Protein and gene circuit level synthetic bioengineering can require years to develop a single target. Phage assisted continuous evolution (PACE) is a powerful new tool for rapidly engineering new genes and proteins, but the method requires an automated cell culture system, making it inaccessible to non industrial research programs. Complex

Protein and gene circuit level synthetic bioengineering can require years to develop a single target. Phage assisted continuous evolution (PACE) is a powerful new tool for rapidly engineering new genes and proteins, but the method requires an automated cell culture system, making it inaccessible to non industrial research programs. Complex protein functions, like specific binding, require similarly dynamic PACE selection that can be alternatively induced or suppressed, with heat labile chemicals like tetracycline. Selection conditions must be controlled continuously over days, with adjustments made every few minutes. To make PACE experiments accessible to the broader community, we designed dedicated cell culture hardware and integrated optogenetically controlled plasmids. The low cost and open source platform allows a user to conduct PACE with continuous monitoring and precise control of evolution using light.

ContributorsTse, Ashley (Author) / Bartelle, Benjamin (Thesis director) / Tian, Xiaojun (Committee member) / Barrett, The Honors College (Contributor) / Materials Science and Engineering Program (Contributor) / School of International Letters and Cultures (Contributor) / Harrington Bioengineering Program (Contributor)
Created2023-05