Journal
NATURE NANOTECHNOLOGY
Volume 15, Issue 6, Pages 500-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41565-020-0662-0
Keywords
-
Funding
- European Research Council (ERC) under the European Union Horizon 2020 research and innovation programme [802567]
- ERC Starting Grant [ERCSTG-2015-680242]
- Zuckerman foundation
- POLAK Fund for Applied Research at the Technion
- Israel Science Foundation [1421/17, 450/18]
Ask authors/readers for more resources
A high-throughput nanoscale 3D microscopy technique, combining imaging flow cytometry with a point-spread function, allows 3D localization measurements in live flowing cells. Capturing the dynamics of live cell populations with nanoscale resolution poses a significant challenge, primarily owing to the speed-resolution trade-off of existing microscopy techniques. Flow cytometry would offer sufficient throughput, but lacks subsample detail. Here we show that imaging flow cytometry, in which the point detectors of flow cytometry are replaced with a camera to record 2D images, is compatible with 3D localization microscopy through point-spread-function engineering, which encodes the depth of the emitter into the emission pattern captured by the camera. The extraction of 3D positions from sub-cellular objects of interest is achieved by calibrating the depth-dependent response of the imaging system using fluorescent beads mixed with the sample buffer. This approach enables 4D imaging of up to tens of thousands of objects per minute and can be applied to characterize chromatin dynamics and the uptake and spatial distribution of nanoparticles in live cancer cells.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available