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Near-infrared branding efficiently correlates light and electron microscopy
Derron Bishop1, Ivana Nikić, Mary Brinkoetter
1Department of Physiology, Indiana University School of Medicine-Muncie, Muncie, Indiana, USA.
Nature Methods
|June 7, 2011
Summary
Near-infrared branding (NIRB) creates 3D fluorescent marks in tissues. These marks allow researchers to correlate light and electron microscopy images, even for tiny structures like dendritic spines.
Area of Science:
- Neuroscience
- Microscopy
- Biotechnology
Background:
- Correlating light and electron microscopy is crucial for understanding complex tissue structures.
- Existing methods for correlating these microscopy techniques are often challenging and limited.
Purpose of the Study:
- To develop a novel method for correlating light and electron microscopy in fixed tissues.
- To enable precise re-identification of microscopic structures across different imaging modalities.
Main Methods:
- Developed near-infrared branding (NIRB) using a pulsed, near-infrared laser.
- Created three-dimensional fiducial marks within fixed tissue samples.
- Utilized the fluorescent properties of the marks for initial imaging and photo-oxidation for electron contrast.
Main Results:
- NIRB successfully generated defined fiducial marks in 3D.
- These marks were fluorescent and could be converted to electron-dense contrast.
- Enabled re-identification of structures as small as dendritic spines between light and electron microscopy.
Conclusions:
- Near-infrared branding (NIRB) is an effective technique for correlating light and electron microscopy.
- This method overcomes a major challenge in high-resolution biological imaging.
- NIRB facilitates detailed structural analysis of complex tissues at multiple scales.
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