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BIOE Seminar: Revealing subcellular structures with live-cell and 3D fluorescence nanoscopy

Speaker Fang Huang, assistant professor, Weldon School of Biomedical Engineering, Purdue University
Date: 2/12/2019
Time: 11:30 a.m.
Location: 2310 Everitt Lab
Sponsor: Bioengineering Department + Big 10 Exchange
Event Type: Seminar/Symposium
 

Abstract

We are in an exciting era of Biomedical Imaging where the inner-workings of cells and tissues can be explored by rapidly developing imaging methods. Labeling specificity and live cell compatibility make fluorescence microscopy an important tool in biomedical research. Its resolution, however, is limited by diffraction to ~250 nm, preventing us from resolving detailed structures within the cell. The recent advent of single molecule switching nanoscopy methods (SMSN, also known as PALM/STORM), overcomes this fundamental limit by stochastically switching single fluorophores on and off so that their emission events can be localized with high precision resulting in a reconstructed image with down to ~25 nm lateral resolution. However, its application has been largely limited to fixed and flat samples due to the poor temporal resolution, inferior resolution in z, and rapidly deteriorating resolution in thick samples. In this talk, I will present some of our most recent developments which synergistically combine novel algorithm designs1,2, newly available sensors/devices3,4 and novel instrumentation5 to allow SMSN imaging in living cells and in 3D. I will demonstrate the capabilities of these new super-resolution systems by revealing subcellular structures from a diverse set of biological systems including virus, bacteria, yeast, mammalian cells and tissue sections.

  1. Zhang, P. et al. Analyzing complex single-molecule emission patterns with deep learning. Nat. Methods 15, 913–916 (2018)
  2. Liu, S. et al. sCMOS noise-correction algorithm for microscopy images. Nat. Methods 14, 760 (2017).
  3. Mlodzianoski, M. J. et al. Active PSF shaping and adaptive optics enable volumetric localization microscopy through brain sections. Nat. Methods 15, pages583–586 (2018)
  4. Huang, F. et al. Video-rate nanoscopy using sCMOS camera-specific single-molecule localization algorithms. Nat. Methods 10, 653–8 (2013).
  5. Huang, F. et al. Ultra-High Resolution 3D Imaging of Whole Cells. Cell 166, 1028–1040 (2016).

 

Biography

The Huang lab aims to develop the next generation high resolution optical microscopy methods, known as super-resolution microscopy or ‘nanoscopy’, that are capable of resolving subcellular structures in three dimensions while monitoring their dynamics in living specimens with nanometer resolution. The group builds novel nanoscopy instruments that combine techniques from engineering and physics such as single molecule fluorescence interference, nanofabrication and adaptive optics and seeks to invent algorithms that take advantages of concepts in mathematics, statistics and signal processing. We aim to significantly push the envelope of high resolution imaging in directions of live cell and tissue imaging, two major roadblocks of modern super-resolution techniques and further allow building dynamic structural models of large protein complexes in live cells and tissues 1-10 nm resolution.

Huang earned his bachelor degree in Physics at the University of Science and Technology of China in 2004 and his doctoral degree in Physics from the University of New Mexico in 2011. Before joining Purdue, Fang Huang was a Brown-Coxe Postdoctoral Fellow in Cell Biology at Yale School of Medicine. Huang received Excellence in Research Awards from Purdue, Maximize Investigator Research Award (MIRA) from NIH, 2016 Young Faculty Award from DARPA.

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Bioengineering

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