title: Pneumatic stimulation of C. elegans mechanoreceptor neurons in a microfluidic trap.
abstract: New tools for applying force to animals, tissues, and cells are critically needed in order to advance the field of mechanobiology, as few existing tools enable simultaneous imaging of tissue and cell deformation as well as cellular activity in live animals. Here, we introduce a novel microfluidic device that enables high-resolution optical imaging of cellular deformations and activity while applying precise mechanical stimuli to the surface of the worm's cuticle with a pneumatic pressure reservoir. To evaluate device performance, we compared analytical and numerical simulations conducted during the design process to empirical measurements made with fabricated devices. Leveraging the well-characterized touch receptor neurons (TRNs) with an optogenetic calcium indicator as a model mechanoreceptor neuron, we established that individual neurons can be stimulated and that the device can effectively deliver steps as well as more complex stimulus patterns. This microfluidic device is therefore a valuable platform for investigating the mechanobiology of living animals and their mechanosensitive neurons.
authors.affiliation: Department of Mechanical Engineering, Stanford University, Stanford, California, USA. mbgoodman@stanford.edu pruitt@stanford.edu and Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA. michael.krieg@icfo.eu.
authors.forename: Adam L
authors.initials: AL
authors.lastname: Nekimken
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA. michael.krieg@icfo.eu.
authors.forename: Holger
authors.initials: H
authors.lastname: Fehlauer
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA. michael.krieg@icfo.eu and Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Gothenburg, Sweden.
authors.forename: Anna A
authors.identifier: 0000-0003-1861-3801
authors.initials: AA
authors.lastname: Kim
authors.affiliation: Department of Electrical Engineering, Stanford University, Stanford, California, USA.
authors.forename: Sandra N
authors.initials: SN
authors.lastname: Manosalvas-Kjono
authors.affiliation: Department of Aeronautics and Astronautics, Stanford University, Stanford, California, USA.
authors.forename: Purim
authors.initials: P
authors.lastname: Ladpli
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, California, USA.
authors.forename: Farah
authors.initials: F
authors.lastname: Memon
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA. michael.krieg@icfo.eu.
authors.forename: Divya
authors.initials: D
authors.lastname: Gopisetty
authors.affiliation: Department of Mechanical Engineering, Stanford University, Stanford, California, USA. mbgoodman@stanford.edu pruitt@stanford.edu.
authors.forename: Veronica
authors.initials: V
authors.lastname: Sanchez
authors.affiliation: Department of Mechanical Engineering, Stanford University, Stanford, California, USA. mbgoodman@stanford.edu pruitt@stanford.edu and Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA. michael.krieg@icfo.eu.
authors.forename: Miriam B
authors.identifier: 0000-0002-5810-1272
authors.initials: MB
authors.lastname: Goodman
authors.affiliation: Department of Mechanical Engineering, Stanford University, Stanford, California, USA. mbgoodman@stanford.edu pruitt@stanford.edu and Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA. michael.krieg@icfo.eu and Department of Bioengineering, Stanford University, Stanford, California, USA.
authors.forename: Beth L
authors.initials: BL
authors.lastname: Pruitt
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA. michael.krieg@icfo.eu.
authors.forename: Michael
authors.identifier: 0000-0003-0501-5036
authors.initials: M
authors.lastname: Krieg
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans; D002118:Calcium / Q000378:metabolism; D004867:Equipment Design; D056656:Lab-On-A-Chip Devices; D008465:Mechanoreceptors / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods; D061848:Optical Imaging; D062308:Optogenetics; D010812:Physical Stimulation / Q000295:instrumentation / Q000379:methods
chemical_list: D002118:Calcium