Name: calcium imaging in freely behaving caenorhabditis elegans with well-controlled, nonlocalized vibration
Uploaded: 2021-04-29T15:00:00
Description: Watch this Scientific Journal Video about Calcium Imaging in Freely Behaving Caenorhabditis elegans with Well-Controlled, Nonlocalized Vibration at JoVE.com

We thank the Caenorhabditis Genetics Center for providing the strains used in this study. This publication was supported by JSPS KAKENHI Grant-in-Aid for Scientific research (B) (Grant no. JP18H02483), on Innovative areas &#34;Science of Soft Robot&#34; project (Grant no. JP18H05474), the PRIME from Japan Agency for Medical Research and Development (grant number 19gm6110022h001), and the Shimadzu foundation.

1. Cultivation of worms until calcium imaging
<ol>
	<li>Four days before a calcium imaging experiment, transfer two adult ST12 worms to a new nematode growth medium (NGM) plate (Table of Materials) on which Escherichia coli OP50 are streaked in a square pattern (approximately 4 mm x 4 mm) using a cell spreader so that the worm spends most of the time in the bacteria during calcium imaging12.</li>
	<li>Incubate this NGM plate for 4 days at 20 &#176;C in an incubator (<st...

<ol>
	<li>Hill, P. S. M., Wessel, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biotremology.">Biotremology.</a> Current Biology. 26 (5), 187-191 (2016).</li><li>Fettiplace, R., Hackney, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+sensory+and+motor+roles+of+auditory+hair+cells.">The sensory and motor roles of auditory hair cells.</a> Nature Reviews Neuroscience. 7 (1), 19-29 (2006).</li><li>Vogel, V., Sheetz, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+force+and+geometry+sensing+regulate+cell+functions.">Local force and geometry sensing regulate cell functions.</a> Nature Reviews Molecular Cell Biology. 7 (4), 265-275 (2006).</li><li>Katta, S., Krieg, M., Goodman, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeling+force:+physical+and+physiological+principles+enabling+sensory+mechanotransduction.">Feeling force: physical and physiological principles enabling sensory mechanotransduction.</a> Annual Review of Cell and Developmental Biology. 31, 347-371 (2015).</li><li>Orr, A. W., Helmke, B. P., Blackman, B. R., Schwartz, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+mechanotransduction.">Mechanisms of mechanotransduction.</a> Developmental Cell. 10 (1), 11-20 (2006).</li><li>Goodman, M. B., Sengupta, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+Caenorhabditis+elegans+senses+mechanical+stress,+temperature,+and+other+physical+stimuli.">How Caenorhabditis elegans senses mechanical stress, temperature, and other physical stimuli.</a> Genetics. 212 (1), 25-51 (2019).</li><li>Chalfie, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neural+circuit+for+touch+sensitivity+in+Caenorhabditis+elegans.">The neural circuit for touch sensitivity in Caenorhabditis elegans.</a> The Journal of Neuroscience: The Official Journal of the Society for Neuroscience. 5 (4), 956-964 (1985).</li><li>Wicks, S. R., Rankin, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+integration+of+antagonistic+reflexes+revealed+by+laser+ablation+of+identified+neurons+determines+habituation+kinetics+of+the+Caenorhabditis+elegans+tap+withdrawal+response.">The integration of antagonistic reflexes revealed by laser ablation of identified neurons determines habituation kinetics of the Caenorhabditis elegans tap withdrawal response.</a> Journal of Comparative Physiology. A Sensory, Neural, and Behavioral Physiology. 179 (5), 675-685 (1996).</li><li>Rankin, C. H., Beck, C. D., Chiba, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Caenorhabditis+elegans:+a+new+model+system+for+the+study+of+learning+and+memory.">Caenorhabditis elegans: a new model system for the study of learning and memory.</a> Behavioural Brain Research. 37 (1), 89-92 (1990).</li><li>Bozorgmehr, T., Ardiel, E. L., McEwan, A. H., Rankin, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+plasticity+in+a+Caenorhabditis+elegans+mechanosensory+circuit.">Mechanisms of plasticity in a Caenorhabditis elegans mechanosensory circuit.</a> Frontiers in Physiology. 4, 88 (2013).</li><li>Sugi, T., Ohtani, Y., Kumiya, Y., Igarashi, R., Shirakawa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+optical+quantification+of+mechanosensory+habituation+reveals+neurons+encoding+memory+in+Caenorhabditis+elegans.">High-throughput optical quantification of mechanosensory habituation reveals neurons encoding memory in Caenorhabditis elegans.</a> Proceedings of the National Academy of Sciences of the United States of America. 111 (48), 17236-17241 (2014).</li><li>Stiernagle, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maintenance+of+C.+elegans.">Maintenance of C. elegans.</a> WormBook. , 1-11 (2006).</li><li>Sugi, T., Okumura, E., Kiso, K., Igarashi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoscale+mechanical+stimulation+method+for+quantifying+C.+elegans+mechanosensory+behavior+and+memory.">Nanoscale mechanical stimulation method for quantifying C. elegans mechanosensory behavior and memory.</a> Analytical Sciences: The International Journal of the Japan Society for Analytical Chemistry. 32 (11), 1159-1164 (2016).</li><li>Brownell, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Compressional+and+surface+waves+in+sand:+used+by+desert+scorpions+to+locate+prey.">Compressional and surface waves in sand: used by desert scorpions to locate prey.</a> Science. 197 (4302), 479-482 (1977).</li><li>Clark, D. A., Gabel, C. V., Gabel, H., Samuel, A. D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+activity+patterns+in+thermosensory+neurons+of+freely+moving+Caenorhabditis+elegans+encode+spatial+thermal+gradients.">Temporal activity patterns in thermosensory neurons of freely moving Caenorhabditis elegans encode spatial thermal gradients.</a> Journal of Neuroscience. 27 (23), 6083-6090 (2007).</li><li>Tsukada, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reconstruction+of+spatial+thermal+gradient+encoded+in+thermosensory+neuron+AFD+in+Caenorhabditis+elegans.">Reconstruction of spatial thermal gradient encoded in thermosensory neuron AFD in Caenorhabditis elegans.</a> Journal of Neuroscience. 36 (9), 2571-2581 (2016).</li><li>Piggott, B. J., Liu, J., Feng, Z., Wescott, S. A., Xu, X. Z. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neural+circuits+and+synaptic+mechanisms+underlying+motor+initiation+in+C.+elegans.">The neural circuits and synaptic mechanisms underlying motor initiation in C. elegans.</a> Cell. 147 (4), 922-933 (2011).</li><li>Nguyen, J. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-brain+calcium+imaging+with+cellular+resolution+in+freely+behaving+Caenorhabditis+elegans.">Whole-brain calcium imaging with cellular resolution in freely behaving Caenorhabditis elegans.</a> Proceedings of the National Academy of Sciences of the United States of America. 113 (8), 1074-1081 (2016).</li><li>Schrodel, T., Prevedel, R., Aumayr, K., Zimmer, M., Vaziri, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain-wide+3D+imaging+of+neuronal+activity+in+Caenorhabditis+elegans+with+sculpted+light.">Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light.</a> Nature Methods. 10 (10), 1013-1020 (2013).</li><li>Prevedel, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+whole-animal+3D+imaging+of+neuronal+activity+using+light-field+microscopy.">Simultaneous whole-animal 3D imaging of neuronal activity using light-field microscopy.</a> Nature Methods. 11, 727-730 (2014).</li><li>Nichols, A. L. A., Eichler, T., Latham, R., Zimmer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+global+brain+state+underlies+C.+elegans+sleep+behavior.">A global brain state underlies C. elegans sleep behavior.</a> Science. 356 (6344), (2017).</li><li>Zheng, M., Cao, P., Yang, J., Xu, X. Z. S., Feng, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+imaging+of+multiple+neurons+in+freely+behaving+C.+elegans.">Calcium imaging of multiple neurons in freely behaving C. elegans.</a> Journal of Neuroscience Methods. 206 (1), 78-82 (2012).</li><li>Sugi, T., Ito, H., Nishimura, M., Nagai, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=C.+elegans+collectively+forms+dynamical+networks.">C. elegans collectively forms dynamical networks.</a> Nature Communications. 10 (1), 1-9 (2019).</li></ol>

Generally, the quantification of neural activity requires introduction of a probe and/or restraints on animal body movement. However, for studies of mechanosensory behavior, the invasive introduction of a probe and restraints themselves constitute mechanical stimuli. C. elegans provides a system to circumvent these problems, because its features are transparent and because it has a simple, compact neural circuit comprising only 302 neurons. Combining these advantages with the previously developed method of evoki...

Animals are often exposed to nonlocalized mechanical stimuli such as vibrations or acoustic waves1,2. Because these stimuli influence homeostasis, development, and reproduction, animals must modify their behaviors to cope with them3,4,5. However, the neural circuits and mechanisms underlying such behavioral modification are poorly understood.
Mechanosensory behavior in the nematode, Caenorhabditis elegans, is a simple behavioral paradigm, in which worms usually change behavior from forward movement to a backward escape response when they encounter nonlocalized vibration6. The neural circuit underlying this behavior is composed primarily of five sensory neurons, four pairs of interneurons, and several types of motor neurons7,8. Additionally, worms habituate to such mechanical stimuli after spaced training involving repeated stimulation9,10,11. Therefore, this simple behavioral response constitutes an ideal system to investigate neural mechanisms underlying both nonlocalized vibration-evoked behavior and memory. A protocol for calcium imaging in freely behaving worms under the influence of nonlocalized vibrations is illustrated. Compared with previously reported systems, this system is simple in that it does not require an additional camera for tracking; however, it allows us to change the frequency, displacement, and duration of nonlocalized vibration. Because activation of the AVA interneurons induces the backward escape response, worms co-expressing GCaMP, a calcium indicator, and TagRFP, a calcium-insensitive fluorescent protein, under the control of an AVA-specific promoter were used as an example (see Table of Materials for details). The protocol demonstrates the activation of AVA neurons as a worm switches from forward to backward movement. This protocol facilitates understanding the neural circuit mechanism underlying mechanosensory behavior.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Data anaylsis software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DualViewImaging.nb</td>
			<td>author</td>
			<td></td>
			<td>For analysis of acquired data</td>
		</tr>
		<tr>
			<td>Mathematica12</td>
			<td>Wolfram</td>
			<td></td>
			<td>For running data anaysis software DualViewImaging</td>
		</tr>
		<tr>
			<td>Escherichia coli and C. elegans strains</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>E. coli OP50</td>
			<td>Caenorhabditis Genetics Center</td>
			<td>OP50</td>
			<td>Food for C. elegans. Uracil auxotroph. E. coli B.</td>
		</tr>
		<tr>
			<td>lite-1(ce314) strain</td>
			<td>Caenorhabditis Genetics Center</td>
			<td>KG1180</td>
			<td>Light-insensitive mutant</td>
		</tr>
		<tr>
			<td>lite-1(ce314) strain expressing NLS-GCaMP-NLS and TagRFP under the control of the AVA-speciric promoter</td>
			<td>author</td>
			<td>ST12</td>
			<td>lite-1(ce314) mutant carrying the genes expressing NLS-GCaMP5G-NLS (NLS; nuclear localization signal) and TagRFP under the control of the flp-18 promoter as an extrachoromosomal arrays</td>
		</tr>
		<tr>
			<td>Laser Doppler vibrometer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Lase Doppler vibrometer</td>
			<td>Polytec Japan</td>
			<td>IVS-500</td>
			<td>For quantifying&#160; frequency and displacement generated by the accoustic transducer</td>
		</tr>
		<tr>
			<td>Mouse macro system</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Assay.txt</td>
			<td>Author</td>
			<td></td>
			<td>Script for temporally and specially controlling mouse cursol in Windows</td>
		</tr>
		<tr>
			<td>HiMacroEx</td>
			<td>Vector</td>
			<td>https://www.vector.co.jp/download/file/winnt/util/fh667310.html</td>
			<td>Free download software for controling mouse cursor based on a script</td>
		</tr>
		<tr>
			<td>Nematode growth media plate</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Agar purified, powder</td>
			<td>Nakarai tesque</td>
			<td>01162-15</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>Bacto pepton</td>
			<td>Becton Dickinson</td>
			<td>211677</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>Calcium chloride</td>
			<td>Wako</td>
			<td>036-00485</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>Cholesterol</td>
			<td>Wako</td>
			<td>034-03002</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>di-Photassium hydrogenphosphate</td>
			<td>Nakarai tesque</td>
			<td>28727-95</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>LB broth, Lennox</td>
			<td>Nakarai tesque</td>
			<td>20066-95</td>
			<td>For culture of E. coli OP50</td>
		</tr>
		<tr>
			<td>Magnesium sulfate anhydrous</td>
			<td>TGI</td>
			<td>M1890</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>Potassium Dihydrogenphosphate</td>
			<td>Nakarai tesque</td>
			<td>28720-65</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Nakarai tesque</td>
			<td>31320-05</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>Petri dishes (60 mm)</td>
			<td>Nunc</td>
			<td>150270</td>
			<td>For preparation of NGM plates</td>
		</tr>
		<tr>
			<td>Nonlocalized vibration device</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Amplifier</td>
			<td>LEPY</td>
			<td>LP-A7USB</td>
			<td>For stimulation with controllable vibration</td>
		</tr>
		<tr>
			<td>Acoustic transducer</td>
			<td>MinebeaMitsumi</td>
			<td>LVC25</td>
			<td>For stimulation with controllable vibration</td>
		</tr>
		<tr>
			<td>WaveGene Ver. 1.5</td>
			<td>Thrive</td>
			<td>http://efu.jp.net/soft/wg/down_wg.html</td>
			<td>Free download software for controling vibration property</td>
		</tr>
		<tr>
			<td>Noninvasive calcium imaging</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2-Channel benchtop 3-phase brushless DC servo controller</td>
			<td>Thorlabs</td>
			<td>BBD202</td>
			<td>Compatible controller for MLS203-1 stages</td>
		</tr>
		<tr>
			<td>479/585 nm BrightLine dual-band bandpass filter</td>
			<td>Semrock</td>
			<td>FF01-479/585-25</td>
			<td>For acquisition of two channel images (GCaMP and TagRFP)</td>
		</tr>
		<tr>
			<td>505/606 nm BrightLine dual-edge standard epi-fluorescence dichroic beamsplitter</td>
			<td>Semrock</td>
			<td>FF505/606-Di01-25x36</td>
			<td>For acquisition of two channel images (GCaMP and TagRFP)</td>
		</tr>
		<tr>
			<td>512/25 nm BrightLine single-band bandpass filter</td>
			<td>Semrock</td>
			<td>FF01-512/25-25</td>
			<td>For acquisition of two channel images (GCaMP and TagRFP)</td>
		</tr>
		<tr>
			<td>630/92 nm BrightLine single-band bandpass filter</td>
			<td>Semrock</td>
			<td>FF01-630/92-25</td>
			<td>For acquisition of two channel images (GCaMP and TagRFP)</td>
		</tr>
		<tr>
			<td>Computer</td>
			<td>Dell</td>
			<td>Precision T7600</td>
			<td>Windows7 with Intel Xeon CPU ES-2630 and 8 GB of RAM</td>
		</tr>
		<tr>
			<td>High-speed x-y motorized stage</td>
			<td>Thorlabs</td>
			<td>MLS203-1</td>
			<td>Fast XY scannning stage</td>
		</tr>
		<tr>
			<td>Image splitting optics</td>
			<td>Hamamatsu photonics</td>
			<td>A12801-01</td>
			<td>For acquisition of two channel images (GCaMP and TagRFP) generated by W-VIEW GEMINI Image spliting optics</td>
		</tr>
		<tr>
			<td>LED light source</td>
			<td>CoolLED</td>
			<td>pE-4000</td>
			<td>For generating 470 nm and 560 nm excitation light</td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Olympus</td>
			<td>MVX10</td>
			<td></td>
		</tr>
		<tr>
			<td>sCMOS camera</td>
			<td>Andor</td>
			<td>Zyla</td>
			<td></td>
		</tr>
		<tr>
			<td>x 2 Objective lens</td>
			<td>Olympus</td>
			<td>MVPLAPO2XC</td>
			<td>Working distance 20 mm and numerical aperture 0.5</td>
		</tr>
		<tr>
			<td>Plasmid</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>pKDK66 plasmid</td>
			<td>author</td>
			<td>pKDK66</td>
			<td>Co-injection marker</td>
		</tr>
		<tr>
			<td>pTAK83 plasmid</td>
			<td>author</td>
			<td>pTAK83</td>
			<td>Plasmid for expression of TagRFP under the control of&#160; the flp-18 promoter</td>
		</tr>
		<tr>
			<td>pTAK144 plasmid</td>
			<td>author</td>
			<td>pTAK144</td>
			<td>Plasmid for expression of NLS-GCaMP5G-NLS under the control of&#160; the flp-18 promoter</td>
		</tr>
		<tr>
			<td>Tracking software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>homingback.vi</td>
			<td>author</td>
			<td></td>
			<td>SubVi file for tracking a fluoresent spot of a worm through feedback control of sCMOS camera and x-y motorized stage</td>
		</tr>
		<tr>
			<td>LabVIEW</td>
			<td>National instruments</td>
			<td></td>
			<td>For running tracking software</td>
		</tr>
		<tr>
			<td>Zyla Control ver.2.6CI.vi</td>
			<td>author</td>
			<td></td>
			<td>For tracking a fluoresent spot of a worm through feedback control of sCMOS camera and x-y motorized stage</td>
		</tr>
	</tbody>
</table>

calcium imaging in freely behaving caenorhabditis elegans with well-controlled, nonlocalized vibration

Nonlocalized mechanical forces, such as vibrations and acoustic waves, influence a wide variety of biological processes from development to homeostasis. Animals cope with these stimuli by modifying their behavior. Understanding the mechanisms underlying such behavioral modification requires quantification of neural activity during the behavior of interest. Here, we report a method for calcium imaging in freely behaving Caenorhabditis elegans with nonlocalized vibration of specific frequency, displacement, and duration. This method allows the production of well-controlled, nonlocalized vibration using an acoustic transducer and quantification of evoked calcium responses at single-cell resolution. As a proof of principle, the calcium response of a single interneuron, AVA, during the escape response of C. elegans to vibration is demonstrated. This system will facilitate understanding of neural mechanisms underlying behavioral responses to mechanical stimuli.

Here, a worm expressing both GCaMP and TagRFP under control of the AVA interneuron-specific promoter is used as an example of calcium imaging in freely behaving C. elegans. GCaMP and TagRFP channel data were obtained as a series of images, some of which are shown in Figure 6 and as a movie (Supplemental Movie 1). The displacement of the Petri plate induced by our nonlocalized vibration system (Figure 7) was also quantified. The displace...

Watch this Scientific Journal Video about Calcium Imaging in Freely Behaving Caenorhabditis elegans with Well-Controlled, Nonlocalized Vibration at JoVE.com

Calcium Imaging in Freely Behaving Caenorhabditis elegans with Well-Controlled, Nonlocalized Vibration

Reported here is a system for calcium imaging in freely behaving Caenorhabditis elegans with well-controlled, nonlocalized vibration. This system allows researchers to evoke nonlocalized vibrations with well-controlled properties at nano-scale displacement and to quantify calcium currents during responses of C. elegans to the vibrations.

Calcium Imaging in Freely Behaving Caenorhabditis elegans with Well-Controlled, Nonlocalized Vibration

Nonlocalized mechanical forces, such as vibrations and acoustic waves, influence a wide variety of biological processes from development to homeostasis. ...

Research

JoVE Journal

Behavior

Functional Magnetic Resonance Imaging (fMRI) with Auditory Stimulation in Songbirds

Functional Magnetic Resonance Imaging fMRI with Auditory Stimulation in Songbirds

The neurobiology of birdsong, as a model for human speech, is a pronounced area of research in behavioral neuroscience. Whereas electrophysiology and ...

Construction of Microdrive Arrays for Chronic Neural Recordings in Awake Behaving Mice

State-of-the-art electrophysiological recordings from the brains of freely behaving animals allow researchers to simultaneously examine local field ...

Long-term Potentiation of Perforant Pathway-dentate Gyrus Synapse in Freely Behaving Mice

Studies of long-term potentiation of synaptic efficacy, an activity-dependent synaptic phenomenon having properties that make it attractive as a potential ...

Two-photon Calcium Imaging in Mice Navigating a Virtual Reality Environment

In recent years, two-photon imaging has become an invaluable tool in neuroscience, as it allows for chronic measurement of the activity of genetically ...

Measurement of Fronto-limbic Activity Using an Emotional Oddball Task in Children with Familial High Risk for Schizophrenia

Adolescence is a critical developmental period where the early symptoms of schizophrenia frequently emerge. First-degree relatives of people with ...

Measurement of Vibration Detection Threshold and Tactile Spatial Acuity in Human Subjects

Tests that allow the precise determination of psychophysical thresholds for vibration and grating orientation provide valuable information about ...

A Novel Experimental and Analytical Approach to the Multimodal Neural Decoding of Intent During Social Interaction in Freely-behaving Human Infants

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental ...

Multi-layer Cortical Ca2+ Imaging in Freely Moving Mice with Prism Probes and Miniaturized Fluorescence Microscopy

Multi-layer Cortical Ca2+ Imaging in Freely Moving Mice with Prism Probes and Miniaturized Fluorescence Microscopy

In vivo circuit and cellular level functional imaging is a critical tool for understanding the brain in action. High resolution imaging of mouse cortical ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

Whole Body Vibration Methods with Survivors of Polio

The purpose of the original study was to examine the use of whole body vibration (WBV) on polio survivors with and without post-polio syndrome as a form ...