Ex Vivo Calcium Imaging to Study Drosophila Brain Responses to Neuropeptides

Abstract

Source: Ishimoto, H. et. al., Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila. J. Vis. Exp. (2018)

This video demonstrates calcium imaging to study Drosophila brain responses to neuropeptides. Neurons expressing a fluorescent calcium indicator reveal intracellular calcium dynamics and functional responses to signaling molecules.

Protocol

1. Preparation of Larval Brain Explants

Results

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Tungsten rod	A-M systems, Sequim, WA, USA	717000
Blu Tack	Bostik, Paris, France	3049100	Putty-like reusable adhesives
Watch glass, square, 1 5/8 in	Carolina Biological Supply Company, Burlington, NC, USA	742300
PBS	TAKARA Bio Inc., Kusatsu, Shiga, Japan	T900	Phosphated buffered salts
CCHa2	SCRUM Inc., Tokyo, Japan		Custum-synthesized peptide
Ghrerin	Peptide institute Inc., Osaka, Japan	4372-s
Nociceptin	Peptide institute Inc., Osaka, Japan	4313-v
Axio Imager A2	Carl Zeiss, Oberkochen, Germany	Axio Imager A2	Fluorescence microscope
Objective W N-Achroplan 20x/0.5 M27	Carl Zeiss, Oberkochen, Germany	420957-9900-000	Water-immersion objective lens
P-725 PIFOC objective scanner with long travel range	Physik Instrumente GmbH & Co. KG, Karlsruhe, Germany	N/A	Piezoelectric-activated lens mover
Confocal scanner unit CSU-W1	Yokogawa Electric Corporation, Tokyo, Japan	CSU-W1	Spinning disc confocal head
ImagEM C9100-13	Hamamatsu Photonics, Sizuoka, Japan	C9100-13	EM-CCD camera
OBIS 488 nm LS 60 mW	Coherent, Santa Clara, CA, USA	1178770	488-nm laser
488/568/647 nm Yokogawa dichroic beamsplitter	Semrock, Rochester, NY, USA	Di01-T405/488/568/647-13x15x0.5	Dichroic beam splitter
528/38 nm BrightLine single-band bandpass filter	Semrock, Rochester, NY, USA	FF01-528/38-25	Emission filter
µManager	Open Imaging, Inc.	N/A	https://micro-manager.org