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Cryofixation during live-imaging enables millisecond time-correlated light and electron microscopy
M Fuest1, G M Nocera1, M M Modena1
1Max Planck Institute for Biophysical Chemistry, Goettingen, Germany.
Journal of Microscopy
|August 9, 2018
Summary
This study introduces in situ cryofixation for millisecond time-correlated live imaging and electron microscopy. This method improves ultrastructural preservation in Caenorhabditis elegans by eliminating sample transfer delays.
Area of Science:
- Cell biology
- Microscopy
- Biophysics
Background:
- Correlating live-cell imaging with electron microscopy links cellular dynamics to ultrastructure.
- Current methods have >1-second delay due to sample transfer for cryofixation.
- This limits the temporal resolution of correlative light and electron microscopy.
Purpose of the Study:
- To develop a method for millisecond time-correlated live imaging and electron microscopy.
- To overcome the temporal limitations of current correlative techniques.
- To enable real-time selection and arrest of biological states for ultrastructural analysis.
Main Methods:
- Developed in situ cryofixation directly within the light microscope field of view.
- Used Caenorhabditis elegans as a model system.
- Performed subsequent transmission electron microscopy (TEM).
Main Results:
- Achieved millisecond time-correlation between live imaging and electron microscopy.
- Eliminated the sample transfer step, reducing time lag to freezing rate.
- Demonstrated well-preserved ultrastructure in cryofixed C. elegans.
Conclusions:
- In situ cryofixation enables high temporal resolution correlative imaging.
- This technique opens new avenues for studying biological processes across length scales.
- Facilitates real-time observation and fixation of dynamic cellular events.
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