PhD Defense by Han-Ching (Joyce) Chiu

Event Details
  • Date/Time:
    • Monday October 21, 2019
      9:00 am - 11:00 am
  • Location: EBB 4029
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Summary Sentence: Tunable elastomeric nano-channels from guided fracture for biopolymer super-resolution imaging

Full Summary: No summary paragraph submitted.

Han-Ching (Joyce) Chiu

BME Ph.D. Defense Presentation

 

Date: October 21, 2019

Time: 9AM EST

Location: EBB 4029

 

Advisor: Dr. Shuichi Takayama

Committee Members: Dr. Philip Santangelo, Dr. Todd Sulchek, Dr. Michael Thouless, Dr. Yifan Liu

 

Title: Tunable elastomeric nano-channels from guided fracture for biopolymer super-resolution imaging

 

The development of a polydimethylsiloxane (PDMS) based micro-, nano-fluidics system compatible with super-resolution imaging to capture extracted chromatin, for epigenetics studies. Native chromatins are DNA folded and tightly coiled around proteins, to access the proteins of interest, chromatins must be unwrapped via linearization. An array of cracks ranging from micron-scale to nano-scale is fabricated by subjecting a two-layer elastomeric materials system to an applied tensile strain. The morphology of the generated surface cracks is determined by the pre-patterned photoresist features in a PDMS substrate and the spin coated thin film on its surface. The depths of the cracks governed the thickness and properties of the spin-coated surface layer, while the locations of crack initiation are determined by pre-patterned features in the bulk substrate. The surface cracks are sealed with a thin PDMS film by plasma bonding.  This creates channels from the surface cracks, where the channel widths can be tuned via an automated stretcher that applies a uniaxial strain. This tunable device can be used to capture biopolymers such as DNA and chromatin. The linearization of captured biopolymers is achieved by a combination of nano-confinement in small conduits and the squeezing hydrodynamic flow which results from the strain being released from the stretcher. Various fluorescent dyes are investigated to overcome the oxygen rich PDMS environment that is not conducive to super-resolution microscopy. Tetrahymena thermophila, a protozoan, is chosen as the model system for chromatin extraction and the protein of interest is labeled with super-resolution compatible dye. Thus, direct super resolution imaging of proteins on biopolymers in the tunable nano-channels is achieved. Visualization of the linearized chromatins pointed towards dispersive segregation for histone deposition. We believe that this linearization platform can be adapted for visualization of other protein modifications and  eventually provide valuable epigenetic information.

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Phd Defense
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  • Created By: Tatianna Richardson
  • Workflow Status: Published
  • Created On: Oct 4, 2019 - 9:26am
  • Last Updated: Oct 4, 2019 - 9:26am