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Three-dimensional Imaging of Bacterial Cells for Accurate Cellular Representations and Precise Protein Localization
Published on: October 29, 2019
Three-dimensional sub-100 nm resolution fluorescence microscopy of thick samples
Manuel F Juette1, Travis J Gould, Mark D Lessard
1Institute for Molecular Biophysics, The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine 04609, USA.
Nature Methods
|May 13, 2008
Summary
Researchers developed a novel light microscope for 3D super-resolution imaging of whole cells. This biplane fluorescence photoactivation localization microscopy (FPALM) achieves nanoscale resolution over several micrometers without sacrificing speed.
Area of Science:
- Cellular biology
- Microscopy
- Biophysics
Background:
- Understanding cellular organization requires high-resolution 3D imaging of entire cells.
- Current super-resolution techniques often face limitations in speed, sensitivity, or imaging depth.
Purpose of the Study:
- To develop a light microscope capable of 3D super-resolution imaging of cellular volumes.
- To achieve nanoscale resolution over several micrometers with high speed and sensitivity.
Main Methods:
- Development of a biplane detection scheme.
- Integration with fluorescence photoactivation localization microscopy (FPALM).
- Achieving translationally invariant resolution.
Main Results:
- Demonstrated 3D super-resolution imaging with 30 x 30 x 75 nm resolution.
- Images were acquired over a depth of several micrometers.
- The method, biplane FPALM, maintained high speed and sensitivity.
Conclusions:
- Biplane FPALM enables unprecedented 3D super-resolution imaging of cellular volumes.
- This technique overcomes previous limitations in speed and depth for nanoscale imaging.
- Facilitates deeper insights into cellular organization and function.
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