scholarly journals Immobilization of C. elegans on cultivation plates by thermoelectric cooling for high-throughput subcellular-resolution microscopy

2021 ◽  
Author(s):  
Yao L. Wang ◽  
Erik L. Jaklitsch ◽  
Noa W. F. Grooms ◽  
Leilani G. Schulting ◽  
Samuel H. Chung
Keyword(s):  
High Throughput ◽  
Animal Movement ◽  
Biological Research ◽  
Nematode Caenorhabditis Elegans ◽  
C Elegans ◽  
Cooling Method ◽  
Subcellular Resolution ◽  
High Resolution Microscopy

Imaging, visual screens, and optical surgery are frequently applied to the nematode Caenorhabditis elegans at subcellular resolution for in vivo biological research. However, these approaches remain low-throughput and require significant manual effort. To improve throughput and enable automation in these techniques, we implement a novel cooling method to immobilize C. elegans directly on their cultivation plate. Previous studies cooled animals in microfluidics or flooded wells to 1-4 C. Counterintuitively, we find that cooling to 5-7 C immobilizes animals more effectively than lower temperatures. At 6 C, animal movement consists of bouts of submicron nose tip movement occurring at a sufficiently low magnitude and frequency to permit clear imaging. We demonstrate the ability to perform subcellular-resolution fluorescence imaging, including 64x magnification 3D image stacks and 2-min long timelapse recordings of the ASJ neuron without blurring from animal motion. We also observe no long-term side effects from cooling immobilization on animal lifespan or fecundity. We believe our cooling method enables high-throughput and high-resolution microscopy with no chemical or mechanical interventions.

scholarly journals Organ-specific, multimodal, wireless optoelectronics for high-throughput phenotyping of peripheral neural pathways

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Woo Seok Kim ◽  
Sungcheol Hong ◽  
Milenka Gamero ◽  
Vivekanand Jeevakumar ◽  
Clay M. Smithhart ◽  
...  
Keyword(s):  
High Throughput ◽  
Low Cost ◽  
Optoelectronic Device ◽  
Antenna System ◽  
Neural Pathways ◽  
Organ Specific ◽  
Coil Antenna ◽  
Sensory Fibers

AbstractThe vagus nerve supports diverse autonomic functions and behaviors important for health and survival. To understand how specific components of the vagus contribute to behaviors and long-term physiological effects, it is critical to modulate their activity with anatomical specificity in awake, freely behaving conditions using reliable methods. Here, we introduce an organ-specific scalable, multimodal, wireless optoelectronic device for precise and chronic optogenetic manipulations in vivo. When combined with an advanced, coil-antenna system and a multiplexing strategy for powering 8 individual homecages using a single RF transmitter, the proposed wireless telemetry enables low cost, high-throughput, and precise functional mapping of peripheral neural circuits, including long-term behavioral and physiological measurements. Deployment of these technologies reveals an unexpected role for stomach, non-stretch vagal sensory fibers in suppressing appetite and demonstrates the durability of the miniature wireless device inside harsh gastric conditions.

scholarly journals Correction: Long-term C. elegans immobilization enables high resolution developmental studies in vivo

Lab on a Chip ◽  
2018 ◽  
Vol 18 (12) ◽  
pp. 1802-1802
Author(s):  
Simon Berger ◽  
Evelyn Lattmann ◽  
Tinri Aegerter-Wilmsen ◽  
Michael Hengartner ◽  
Alex Hajnal ◽  
...  
Keyword(s):  
High Resolution ◽  
Developmental Studies ◽  
C Elegans

Correction for ‘Long-term C. elegans immobilization enables high resolution developmental studies in vivo’ by Simon Berger et al., Lab Chip, 2018, 18, 1359–1368.

scholarly journals Long-term C. elegans immobilization enables high resolution developmental studies in vivo

Lab on a Chip ◽  
2018 ◽  
Vol 18 (9) ◽  
pp. 1359-1368 ◽  
Author(s):  
Simon Berger ◽  
Evelyn Lattmann ◽  
Tinri Aegerter-Wilmsen ◽  
Michael Hengartner ◽  
Alex Hajnal ◽  
...  
Keyword(s):  
High Resolution ◽  
Developmental Studies ◽  
Developmental Processes ◽  
C Elegans

Microfluidics enables the interference free observation of sensitive developmental processes in C. elegans.

scholarly journals A calibrated optogenetic toolbox of stable zebrafish opsin lines

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Paride Antinucci ◽  
Adna Dumitrescu ◽  
Charlotte Deleuze ◽  
Holly J Morley ◽  
Kristie Leung ◽  
...  
Keyword(s):  
High Throughput ◽  
Neural Activity ◽  
Ion Selectivity ◽  
Transgenic Zebrafish ◽  
Substantial Variation ◽  
Opsin Expression ◽  
Electrophysiological Recordings ◽  
Subcellular Resolution ◽  
Kinetics Of

Optogenetic actuators with diverse spectral tuning, ion selectivity and kinetics are constantly being engineered providing powerful tools for controlling neural activity with subcellular resolution and millisecond precision. Achieving reliable and interpretable in vivo optogenetic manipulations requires reproducible actuator expression and calibration of photocurrents in target neurons. Here, we developed nine transgenic zebrafish lines for stable opsin expression and calibrated their efficacy in vivo. We first used high-throughput behavioural assays to compare opsin ability to elicit or silence neural activity. Next, we performed in vivo whole-cell electrophysiological recordings to quantify the amplitude and kinetics of photocurrents and test opsin ability to precisely control spiking. We observed substantial variation in efficacy, associated with differences in both opsin expression level and photocurrent characteristics, and identified conditions for optimal use of the most efficient opsins. Overall, our calibrated optogenetic toolkit will facilitate the design of controlled optogenetic circuit manipulations.

Caenorhabditis elegans Learning and Memory

2019 ◽  
Author(s):  
James S.H. Wong ◽  
Catharine H. Rankin
Keyword(s):  
Carbon Dioxide ◽  
Caenorhabditis Elegans ◽  
Classical Conditioning ◽  
Learning And Memory ◽  
Long Term Memory ◽  
Context Conditioning ◽  
Term Memory ◽  
C Elegans

The nematode, Caenorhabditis elegans (C. elegans), is an organism useful for the study of learning and memory at the molecular, cellular, neural circuitry, and behavioral levels. Its genetic tractability, transparency, connectome, and accessibility for in vivo cellular and molecular analyses are a few of the characteristics that make the organism such a powerful system for investigating mechanisms of learning and memory. It is able to learn and remember across many sensory modalities, including mechanosensation, chemosensation, thermosensation, oxygen sensing, and carbon dioxide sensing. C. elegans habituates to mechanosensory stimuli, and shows short-, intermediate-, and long-term memory, and context conditioning for mechanosensory habituation. The organism also displays chemotaxis to various chemicals, such as diacetyl and sodium chloride. This behavior is associated with several forms of learning, including state-dependent learning, classical conditioning, and aversive learning. C. elegans also shows thermotactic learning in which it learns to associate a particular temperature with the presence or absence of food. In addition, both oxygen preference and carbon dioxide avoidance in C. elegans can be altered by experience, indicating that they have memory for the oxygen or carbon dioxide environment they were reared in. Many of the genes found to underlie learning and memory in C. elegans are homologous to genes involved in learning and memory in mammals; two examples are crh-1, which is the C. elegans homolog of the cAMP response element-binding protein (CREB), and glr-1, which encodes an AMPA glutamate receptor subunit. Both of these genes are involved in long-term memory for tap habituation, context conditioning in tap habituation, and chemosensory classical conditioning. C. elegans offers the advantage of having a very small nervous system (302 neurons), thus it is possible to understand what these conserved genes are doing at the level of single identified neurons. As many mechanisms of learning and memory in C. elegans appear to be similar in more complex organisms including humans, research with C. elegans aids our ever-growing understanding of the fundamental mechanisms of learning and memory across the animal kingdom.

scholarly journals Biological effects, translocation, and metabolism of quantum dots in the nematode Caenorhabditis elegans

2016 ◽  
Vol 5 (4) ◽  
pp. 1003-1011 ◽  
Author(s):  
Dayong Wang
Keyword(s):  
Quantum Dots ◽  
Caenorhabditis Elegans ◽  
Recent Progress ◽  
Biological Effects ◽  
Assay System ◽  
Nematode Caenorhabditis Elegans ◽  
C Elegans

We summarize recent progress on the biological effects, translocation, and metabolism of QDs in thein vivoassay system ofC. elegans.

scholarly journals Salmonella biofilms program innate immunity for persistence in Caenorhabditis elegans

2019 ◽  
Vol 116 (25) ◽  
pp. 12462-12467 ◽  
Author(s):  
Stuti K. Desai ◽  
Anup Padmanabhan ◽  
Sharvari Harshe ◽  
Ronen Zaidel-Bar ◽  
Linda J. Kenney
Keyword(s):  
Caenorhabditis Elegans ◽  
Temporal Dynamics ◽  
Innate Immune ◽  
Heterologous Host ◽  
C Elegans ◽  
Parasitic Lifestyle ◽  
Induced Changes ◽  
Infectious Form

The adaptive in vivo mechanisms underlying the switch in Salmonella enterica lifestyles from the infectious form to a dormant form remain unknown. We employed Caenorhabditis elegans as a heterologous host to understand the temporal dynamics of Salmonella pathogenesis and to identify its lifestyle form in vivo. We discovered that Salmonella exists as sessile aggregates, or in vivo biofilms, in the persistently infected C. elegans gut. In the absence of in vivo biofilms, Salmonella killed the host more rapidly by actively inhibiting innate immune pathways. Regulatory cross-talk between two major Salmonella pathogenicity islands, SPI-1 and SPI-2, was responsible for biofilm-induced changes in host physiology during persistent infection. Thus, biofilm formation is a survival strategy in long-term infections, as prolonging host survival is beneficial for the parasitic lifestyle.

scholarly journals Automated Platform for Long-Term Culture and High-Content Phenotyping of Single C. elegans Worms

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
H. B. Atakan ◽  
R. Xiang ◽  
M. Cornaglia ◽  
L. Mouchiroud ◽  
E. Katsyuba ◽  
...  
Keyword(s):  
High Throughput ◽  
Agar Plate ◽  
Model Organism ◽  
Suitable Model ◽  
Microfluidic Chips ◽  
Long Term Culture ◽  
C Elegans ◽  
Bleaching Procedure ◽  
Automated Platform

Abstract The nematode Caenorhabditis elegans is a suitable model organism in drug screening. Traditionally worms are grown on agar plates, posing many challenges for long-term culture and phenotyping of animals under identical conditions. Microfluidics allows for ‘personalized’ phenotyping, as microfluidic chips permit collecting individual responses over worms’ full life. Here, we present a multiplexed, high-throughput, high-resolution microfluidic approach to culture C. elegans from embryo to the adult stage at single animal resolution. We allocated single embryos to growth chambers, for observing the main embryonic and post-embryonic development stages and phenotypes, while exposing worms to up to 8 different well-controlled chemical conditions. Our approach allowed eliminating bacteria aggregation and biofilm formation-related clogging issues, which enabled us performing up to 80 hours of automated single worm culture studies. Our microfluidic platform is linked with an automated phenotyping code that registers organism-associated phenotypes at high-throughput. We validated our platform with a dose-response study of the anthelmintic drug tetramisole by studying its influence through the life cycle of the nematodes. In parallel, we could observe development effects and variations in single embryo and worm viability due to the bleaching procedure that is standardly used for harvesting the embryos from a worm culture agar plate.

C. elegans as robust, high throughput in vivo system for hazard assessment

2015 ◽  
Vol 238 (2) ◽  
pp. S53
Author(s):  
M. Wildwater ◽  
E. Kerkhof ◽  
C. Lokman ◽  
R. Pieters
Keyword(s):  
High Throughput ◽  
Hazard Assessment ◽  
C Elegans
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