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A microfluidics-based method for measuring neuronal activity in Drosophila chemosensory neurons

Nature Protocols volume 11, pages 2389–2400 (2016)Cite this article

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Abstract

Monitoring neuronal responses to defined sensory stimuli is a powerful and widely used approach for understanding sensory coding in the nervous system. However, providing precise, stereotypic and reproducible cues while concomitantly recording neuronal activity remains technically challenging. Here we describe the fabrication and use of a microfluidics system that allows precise temporally restricted stimulation of Drosophila chemosensory neurons with an array of different chemical cues. The system can easily be combined with genetically encoded calcium sensors, and it can measure neuronal activity at single-cell resolution in larval sense organs and in the proboscis or leg of the adult fly. We describe the design of the master mold, the production of the microfluidic chip and live imaging using the calcium sensor GCaMP, expressed in distinct types of Drosophila chemosensory neurons. Fabrication of the master mold and microfluidic chips requires basic skills in photolithography and takes ∼2 weeks; the same devices can be used repeatedly over several months. Flies can be prepared for measurements in minutes and imaged for up to 1 h.

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Figure 1: Overview of workflow, the design of the microfluidic chip and the installation of the pumps and chip on the microscope.
Figure 2: Measurements with larval preparations.
Figure 3: Measurements using adult preparations.

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Acknowledgements

We thank the Bloomington Drosophila Stock Center for reagents. We thank R. Benton, P. Renaud, L. Pethö, K. Suter and J. Dorsaz for help with the experiments, and T. Graham and B. Egger for helpful discussion of the manuscript. This work was supported by grants from the Swiss National Science Foundation (CRSII3_136307 and 31003A_149499) and the European Research Council (ERC-2012-StG 309832-PhotoNaviNet) to S.G.S. We further thank our colleagues in the Sprecher laboratory for fruitful discussions of the manuscript.

Author information

Author notes

  1. Lena van Giesen and G Larisa Neagu-Maier: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biology, Institute of Zoology, University of Fribourg, Fribourg, Switzerland

    Lena van Giesen, G Larisa Neagu-Maier & Simon G Sprecher

  2. Department of Biology, Brandeis University, Waltham, Massachusetts, USA

    Lena van Giesen

  3. Department of Biological Sciences, Sungkyunkwan University, Suwon, Republic of Korea

    Jae Young Kwon

Authors
  1. Lena van Giesen
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  2. G Larisa Neagu-Maier
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  3. Jae Young Kwon
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  4. Simon G Sprecher
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Contributions

L.v.G. and G.L.N.-M. performed the experiments. L.v.G. and S.G.S. developed the protocol. L.v.G., G.L.N.-M., J.Y.K. and S.G.S. wrote the paper.

Corresponding authors

Correspondence to Jae Young Kwon or Simon G Sprecher.

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The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Correction of artifacts in the graph.

(a) Representative fluorescent trace before and after movement artifact correction. The fluorescence intensity value before the moving frame was duplicated to mask the artifact. (b) In case of excessive movement the same procedure cannot be applied, and it is recommended to discard the recording.

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1 Correction of artifacts in the graph. (a,b) Representative fluorescence trace before and after movement artifact correction. (a) The fluorescence intensity value before the moving frame was duplicated to mask the artifact. (b) In case of excessive movement, the same procedure cannot be applied, and it is recommended that the recording be discarded. (PDF 290 kb)

Demonstration of larval dissection and installation in the microfluidic chip. (MOV 28062 kb)

Correction of movement artifacts. (MOV 945 kb)

Supplementary Data

Design of the photomask used for the microfluidic chip fabrication (.cif file). (ZIP 1032 kb)

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van Giesen, L., Neagu-Maier, G., Kwon, J. et al. A microfluidics-based method for measuring neuronal activity in Drosophila chemosensory neurons. Nat Protoc 11, 2389–2400 (2016). https://doi.org/10.1038/nprot.2016.144

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