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In Vivo Two-Photon Imaging of the Olfactory System in Insects.

Marco Paoli1,2, Albrecht Haase3

  • 1Neuroscience Paris-Seine - Institut de Biologie Paris-Seine, Sorbonne Université, INSERM, CNRS, Paris, France. marco.paoli@sorbonne-universite.fr.

Methods in Molecular Biology (Clifton, N.J.)
|April 18, 2025
PubMed
Summary

This chapter details using two-photon neuroimaging for studying insect olfactory systems in vivo. It offers a comprehensive protocol for functional brain imaging, including preparation, labeling, and data analysis techniques.

Keywords:
Antennal lobeCalcium imagingDrosophilaHoney beeInsectsOlfactionTwo-photon microscopy

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Area of Science:

  • Neuroscience
  • Zoology
  • Microscopy

Background:

  • The insect olfactory system is crucial for survival and behavior.
  • Studying neural circuits in vivo requires advanced imaging techniques.
  • Two-photon neuroimaging offers high resolution and deep tissue penetration.

Purpose of the Study:

  • To provide a comprehensive protocol for in vivo two-photon neuroimaging of the insect olfactory system.
  • To guide researchers in selecting and setting up appropriate two-photon microscopy equipment.
  • To illustrate methods for animal preparation, tissue labeling, and data analysis.

Main Methods:

  • Detailed protocol for two-photon laser scanning microscopy in insects.
  • Guidance on selecting and configuring two-photon microscopes.
  • Methods for in vivo preparation and brain tissue labeling.
  • Overview of image processing and data analysis techniques.

Main Results:

  • Demonstration of successful in vivo functional imaging of the insect olfactory system.
  • Acquisition of morphological information from the living insect brain.
  • Practical examples of applying two-photon neuroimaging to olfactory research.

Conclusions:

  • Two-photon neuroimaging is a powerful tool for studying insect olfactory circuits in vivo.
  • The provided protocol facilitates the application of this technique in neuroscience research.
  • This method enables detailed investigation of neural activity and brain structure.