BME Student Seminar: Austin Lefebvre and Hinesh Patel

Austin Lefebvre: Tracking Mitochondria Reveals Metabolic and Dynamic Heterogeneity in Breast Cancers

Abstract: Mitochondria are the powerhouses of the cell, but the difficulty of segmenting, tracking and analyzing individual mitochondria has led to a bottleneck in discovering important biological insights about these complex organelles. Here, we introduce Mitometer, an automated software for unbiased segmentation and tracking of mitochondria in live cell 2D and 3D fluorescence time-lapse images. We multiplex Mitometer with FLIM of NADH to assess metabolic changes between five breast cancer cell lines of varying invasiveness: 3 triple-negative (TNBC), 2 receptor-positive (ER/PR+), and one normal breast epithelial cell line. Additionally, we run a comprehensive 2D and 3D analysis of mitochondria in a large panel of the aforementioned cell lines, in addition to two TNBC patient-derived xenograft models (HCI-010, HCI-002), and normal breast epithelial cells from reduction mammoplasties of five different individuals. We show large variations in many mitochondrial parameters and show the ability to predict whether a mitochondrion belongs to an ER/PR+ cancer cell or a TNBC cell. Mitometer and these results will further the understanding of mitochondria as drivers of complex diseases.
 

Hinesh Patel: Engineering Scalable Multiplexed Molecular Screening Platforms

Abstract: A central role of biomedical research is capturing and evaluating the rich information from genes, proteins and/or other small molecules to provide actionable information. With modern advances and understanding, focus has shifted to sensing multiple analytes in parallel. Yet, further improvements in multiplexed tools are needed for improved throughput. Here we present two technologies that expand our toolbox for multiplexed molecular screening. First, we demonstrate an imaging platform, leveraging the phasor approach to FLIM and a deep learning classifier, to resolve up to four exogenous molecular probes. Resolution is achieved within a single spectral window, adding an orthogonal dimension with which to multiplex fluorescence imaging. Second is an inexpensive and scalable microfluidic platform coupled with novel phase-change gating technology to generate all pair-wise combinations between two sets of reactants, enabling improved multiplexed screening of sample-reagent combinations. The use of droplet microfluidics provides a sharp 30-fold reduction in reagent consumption and nearly 40-fold reduction in liquid handling, suitable for a wide range of screening efforts from medicine to agriculture.