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Using laser scanning confocal microscopy as a guide for electron microscopic study: a simple method for correlation
X J Sun1, L P Tolbert, J G Hildebrand
1ARL Division of Neurobiology, University of Arizona, Tucson 85721.
Summary
This study introduces a simple method combining laser scanning confocal microscopy and electron microscopy to study neural synaptic connections. This technique efficiently correlates 3D light microscopy with electron microscopy details for neuroanatomy research.
Area of Science:
- Neuroscience
- Cell Biology
- Microscopy
Background:
- Studying synaptic connections in the nervous system is challenging due to neuron complexity.
- Electron microscopy offers high detail but correlating it with light microscopy is time-consuming.
Purpose of the Study:
- To develop a simple, reliable method for correlating 3D light microscopy with electron microscopic detail of synaptic relationships.
- To facilitate the study of neuronal connections in the antennal lobe of Manduca sexta.
Main Methods:
- Intracellular labeling of neurons with neurobiotin or biocytin.
- Processing tissue with streptavidin-nanogold for electron microscopy and streptavidin-Cy3 for confocal microscopy.
- Imaging labeled neurons in embedded blocks using laser scanning confocal microscopy before thin-sectioning for electron microscopy.
Main Results:
- A straightforward protocol was established to combine light and electron microscopy.
- The method allows for precise correlation of 3D confocal images with ultrastructural details.
Conclusions:
- This combined microscopy approach simplifies the study of synaptic connections.
- The technique is valuable for neuroanatomical research, particularly in complex neuropils like the antennal lobe.