@article{High-Throughput:3618,
      recid = {3618},
      author = {Lin, Jing},
      title = {High-Throughput Microfluidic Platform for Ultrasensitive  Single-Cell Protein Analyses},
      publisher = {The University of Chicago},
      school = {Ph.D.},
      address = {2021-12},
      pages = {85},
      abstract = {In the wake of heterogeneous nature of most biological  systems, single-cell analysis has gained enormous  importance and popularity in the field of biology, medicine  and even microbiology. Traditional population-averaged  analysis offers good information on the system level,  however at the price of missing out cell-to-cell  variations, which are critical information for building  disease models, understanding drug resistance, or  identifying cancer stem cells. Though single-cell  transcriptomic technologies are well established,  single-cell proteomic platforms seem to be lagging behind.  This is mostly due to the low-level protein quantities  present in single cells and the difficulty to amplify  protein molecules for end-point detection. In this thesis,  I will report microfluidic solutions for ultra-sensitive  single-cell endogenous protein and enzymatic activities  analysis. I will focus on the design principles of the  microfluidic platforms, working pipeline validation and  biological application demonstrations.	Firstly, I will  demonstrate an ultrasensitive single-cell protein  quantification platform based on a valve-based microfluidic  chip designed and fabricated in house. I will show how the  microfluidic platform improves the digital proximity  ligation assay (digital PLA) performance, thus enabling  quantification of rare protein species in single cells.  Then, the assay will be extended to simultaneously quantify  protein and mRNA copy numbers from the same single  cell.
	Then, the same microfluidic platform has been  adapted to profile multiplexed enzymatic activities in  single cells. By combining family-wide chemical probes and  PLA, the assay - named single-cell activity-dependent  proximity ligation (scADPL) – quantifies active enzymatic  molecules of targeted protein species (instead of total  molecules) in single cells. The proof-of-concept is then  demonstrated with 6-plex measurements in cancer cell lines,  and the assay has been applied to characterize cancer  organoids derived from various patient samples.},
      url = {http://knowledge.uchicago.edu/record/3618},
      doi = {https://doi.org/10.6082/uchicago.3618},
}