Bo Huang
University of California-San Francisco, CA·Biophysics
The human genome encodes roughly 20,000 proteins, and for most of them we still do not fully know where they sit inside a cell, what they interact with, or how they move over time. Answering these questions in living cells, without disturbing the proteins' normal behavior, is a central challenge for cell biology.
- Fields
Biophysics
Biology
Biomedical Engineering
- Topics
live-cell imaging
light-sheet microscopy
eSPIM
- Grants
- 1 known grant, through Aug 2027
What they work on
- Develop epi-illumination selective plane illumination microscopy (eSPIM), which lights only a thin plane of the sample, into a live-cell platform for high-resolution, long-term imaging with low photobleaching and phototoxicity.
- Build software packages for eSPIM instrument control and image processing so that other labs can easily adopt the microscope and analyze movies of protein dynamics quantitatively.
- Screen the OpenCell library of about 1,300 endogenously tagged human proteins with eSPIM to find proteins whose movements or localization change unusually through the cell cycle.
- Extend the FP11 tag and CRISPR/Cas9 editing pipeline, which labels proteins in their own genes, as a base for proteome-wide studies of protein localization and interaction networks.
More about the research
In earlier work, the team developed FP11 tags, small labels that can be attached to a protein directly in its own gene. They combined these tags with an efficient CRISPR/Cas9 gene-editing pipeline that edits many genes in parallel. Together, these tools produced the OpenCell library, a collection of human cell lines covering about 1,300 tagged proteins, which the team characterized with imaging and proteomics (large-scale protein measurement). Two problems remain: filming protein dynamics in live cells over long periods, and turning those movies into numbers. This renewal proposes to develop epi-illumination selective plane illumination microscopy (eSPIM) into a broadly usable live-cell imaging platform. The method illuminates only a thin slice of the sample, which supports high-resolution, long-term imaging with less photobleaching (fading of the label) and less light damage to cells. The team will also write software for instrument control and image processing so other labs can adopt the system. To show what the platform can do, they will screen the OpenCell library for proteins that behave unusually as cells move through the cell cycle.
Why it matters
The project would give biologists microscopy and software tools to watch native, unaltered human proteins move inside living cells for long periods. This could support proteome-wide maps of where proteins are located and how they interact in their natural cellular setting. Because the instrument and software are designed for adoption by other labs, researchers studying many different proteins could use them.
Known grants
From NIH and NSF research grants. Labs often have other funding we can't see.
Closest labs
William E Moerner
Stanford University, CA·Biophysics
Develop a 3D super-resolution microscope that images over a large depth range using pupil-plane phase patterns and a tilted light sheet, for single-molecule imaging and tracking in living cells.
Comert Kural
Ohio State University, OH·Biophysics
Build and iteratively optimize a 3D TIRF-SIM microscope, combining surface-limited illumination, patterned-light super-resolution and evanescent field nanometry, for the best spatial and temporal resolution.
Elizabeth Egan
Stanford University, CA·Microbiology & Immunology
Determine which precise steps of Plasmodium falciparum invasion of human red blood cells require the surface proteins CD44 and CD55, using live-cell imaging and advanced microscopy of CRISPR-edited cultured red blood cells.
The school
University of California-San Francisco: 89 labs on PI Pages, $2.1B a year on research, 164 research doctorates a year.
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