BME PhD Prospectus Defense - Kai Chih Huang

  • Starts: 2:00 pm on Thursday, November 29, 2018

Title: “Single-cell metabolic analysis by multiplex stimulated Raman scattering (SRS) imaging cytometry”

Committee: Ji-Xin Cheng, PhD – BME/ECE (Advisor, Chair) Jerome Mertz, PhD – BME Wilson Wong, PhD – BME Lei Tian, PhD – ECE

Abstract: Therefore, single cell analysis is essential to understand cellular heterogeneity, such as tumor heterogeneity and drug-resistant bacterial heterogeneity. Conventionally fluorescence-activated cell sorting has been widely used in the field of immuno-phenotyping with high-throughput. However, the labeling method may perturb the cellular function, especially for tagging small molecules. Stimulated Raman scattering (SRS) microscopy, as a label-free imaging approach with chemical sensitivity, has overcome this limitation and been used to study metabolism at single cell level. However, current SRS imaging is not implemented to address the cellular heterogeneity due to the low throughput. Statistical analysis of a large number of cells (hundreds or thousands of cells) through cytometry with high-throughput around 100 cells per second is needed. For this reason, we are creating a high-speed hyperspectral SRS image cytometry platform based on multiplex excitation, which acquires a Raman spectrum in 5 microsecond. For current multiplex SRS imaging, spectral resolution and sensitivity are both essential to differentiate weak and overlaid Raman peak in the fingerprint region. In order to improve the spectral resolution, we will apply spectral compressor without the trade-off with sensitivity and demonstrate fast determination of antimicrobial susceptibility as an application. In order to improve the sensitivity, we will propose an electronic resonant multiplex SRS image cytometry to further enhance the Raman cross-section of cytochrome c for probing metabolic states of mitochondrial. This platform opens the opportunity for antimicrobial susceptibility testing and for studying cancer metabolism at single cell level.

44 Cummington Mall, room 401

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