ePoster

Combining electrophysiology, tissue clearing, and light sheet microscopy for an integrated approach towards brain circuit understanding

Sebastian Böhm, Marta Solano Mateos, James A. Oakes, Saiedeh Saghafi, Christoph F. Fuchssteiner, Thomas Klausberger, Hans-Ulrich Dodt
FENS Forum 2024(2024)
Messe Wien Exhibition & Congress Center, Vienna, Austria

Conference

FENS Forum 2024

Messe Wien Exhibition & Congress Center, Vienna, Austria

Resources

Authors & Affiliations

Sebastian Böhm, Marta Solano Mateos, James A. Oakes, Saiedeh Saghafi, Christoph F. Fuchssteiner, Thomas Klausberger, Hans-Ulrich Dodt

Abstract

Traditional histological techniques, which require imaging of individual tissue sections, are widely used but tend to be error-prone, time-consuming, and can lead to section loss or non-uniform deformation. Optical sectioning provides a faster and simpler alternative. For instance, light sheet fluorescence microscopy enables the production of high-resolution images of entire brains. This project aimed to use light sheet fluorescence microscopy as a complementary technique to visualize juxtacellular recorded and labeled neurons in anesthetized rats. Our novel approach involved labeling a single neuron in a rat brain with neurobiotin using the juxtacellular recording and labeling technique. Afterwards, we cleared the brain using an innovative 3DISCO protocol and subsequently imaged using light sheet microscopy. Following imaging, the neuron was three-dimensionally reconstructed with Amira software for comprehensive analysis, measurement, and 3D visualization. Our preliminary results introduce a novel clearing protocol designed specifically for rat brains, representing the fusion of single cell labeling with behavioural assays and tissue clearing. To conclude, this study represents a novel technical approach. It uniquely integrates electrophysiology with a well-known optical sectioning imaging method, the so-called light sheet fluorescence microscopy. This methodology not only improves the efficiency and precision of whole-brain imaging but also allows for detailed neuronal analysis and visualization, contributing to our comprehension of brain function and structure.

Unique ID: fens-24/combining-electrophysiology-tissue-2edcafcf