Tools Development and Open Source Neuroscience
Author: Francisco Joaquín Tassara | Email: ftassara@leloir.org.ar
Francisco Tassara1°, Lourdes Simó1°, Mailen Folgueira 1°, Julian Gargiulo2°, María Fernanda Ceriani1°
1° Fundación Instituto Leloir—IIBBA—CONICET
2° Instituto de Nanosistemas, Universidad Nacional de San Martín
In most animals, daily rhythms of activity and rest are regulated by a circadian system. In Drosophila melanogaster, a specific group of neurons known as small ventral lateral neurons play a key role in generating these rhythms. Our research group observed the circadian remodeling of clock neuron terminals, an indication of synaptic plasticity. However, these studies were performed using fluorescence confocal microscopy techniques in fixed brains, which restricts the ability to observe this process dynamically. This poses a challenge in achieving a precise description of the structural changes that occur during the circadian cycle.
In this work we utilize an oblique plane version of light sheet microscopý. This technique utilizes a single objective for both illumination and imaging. The approach has several advantages over confocal microscopy. It minimizes phototoxic damage to the tissue, enabling prolonged time-lapse observations and it allows the use of high numerical aperture objectives, leading to sub-micron spatial resolutions. Here, we demonstrate an upright design of SOLS microscope compatible with imaging a live fly and present the first images of s-LNvs neurons. These advances contribute towards the goal of achieving long-term monitoring of structural remodelling in a single live fly, a key step for the understanding of the biological mechanisms underlying circadian structural plasticity.