Non-microfluidic methods for imaging live C. elegans

Luke, Clifford; Niehaus, Jason Z.; O’Reilly, Linda P.; Watkins, Simon C.

doi:10.1016/j.ymeth.2014.05.002

Cited by 15 publications

(16 citation statements)

References 13 publications

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“…sealing the coverslip while using PSB [ 22 ] 3.) using hydrogel [ 55 , 56 , 57 , 58 ] or 4.) using whichever microfluidics platform is reasonable and can be obtained for your study [ 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 ].…”

Section: Resultsmentioning

confidence: 99%

Stress and timing associated with Caenorhabditis elegans immobilization methods

Manjarrez

Mailler

2020

Heliyon

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Background Caenorhabditis elegans is a model organism used to study gene, protein, and cell influence on function and behavior. These studies frequently require C. elegans to be immobilized for imaging or laser ablation experiments. There are a number of known techniques for immobilizing worms, but to our knowledge, there are no comprehensive studies of the various agents in common use today. New method This study determines the relationship between concentration, immobilization time, exposure time, and recovery likelihood for several immobilization agents. The agents used in this study are 1-Phenoxy-2-propanol, levamisole, sodium azide, polystyrene beads, and environmental cold shock. These tests are conducted using a humidified chamber to keep chemical concentrations consistent. Each of these agents is also tested to determine if they exhibit stress-related after effects using the gcs-1 , daf-16 , hsp-4 , hif-1 , hsp-16.2, and tmem-135 stress reporters. Results We present a range of quick mount immobilization and recovery conditions for each agent tested. This study shows that, under controlled conditions, 1-Phenoxy-2-propanol shows significant stress from the daf-16 reporter. While 1-Phenoxy-2-propanol and sodium azide both create stress related after effects with long term recovery in the case of the hsp-16.2 reporter. Comparison with existing method(s) This study shows that commonly used concentrations of immobilizing agents are ineffective when evaporation is prevented. Conclusions To improve reproducibility of results it is essential to use consistent concentrations of immobilizing agents. It is also critically important to account for stress-related after effects elicited by immobilization agents when designing any experiment.

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Section: Resultsmentioning

confidence: 99%

Stress and timing associated with Caenorhabditis elegans immobilization methods

Manjarrez

Mailler

2020

Heliyon

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“…The transparent nematodes can be easily visualized and enables live imaging of host-pathogen interactions [64,65]. Typically worms are anesthetisized using sodium azide and placed on an agarose pad containing a drop of M9 buffer.…”

Section: Tools and Techniquesmentioning

confidence: 99%

Caenorhabditis elegans as a Model Host to Monitor the Candida Infection Processes

Elkabti

Issi

Rao

2018

JoF

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C. elegans has several advantages as an experimental host for the study of infectious diseases. Worms are easily maintained and propagated on bacterial lawns. The worms can be frozen for long term storage and still maintain viability years later. Their short generation time and large brood size of thousands of worms grown on a single petri dish, makes it relatively easy to maintain at a low cost. The typical wild type adult worm grows to approximately 1.5 mm in length and are transparent, allowing for the identification of several internal organs using an affordable dissecting microscope. A large collection of loss of function mutant strains are readily available from the C. elegans genetic stock center, making targeted genetic studies in the nematode possible. Here we describe ways in which this facile model host has been used to study Candida albicans, an opportunistic fungal pathogen that poses a serious public health threat.

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“…Two principal approaches can be used to observe motile animals over long time scales. The first approach is following the animal during movement using an automated stage or camera [2][3][4][5][6][7][8] and the second is to restrict the movement to a range that is at least as small as the field of view of the camera [9][10][11][12][13][14] . Both methods have their advantages.…”

Section: Introductionmentioning

confidence: 99%

Agarose Microchambers for Long-term Calcium Imaging of <em>Caenorhabditis elegans</em>

Turek

Besseling

Bringmann

2015

JoVE

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Behavior is controlled by the nervous system. Calcium imaging is a straightforward method in the transparent nematode Caenorhabditis elegans to measure the activity of neurons during various behaviors. To correlate neural activity with behavior, the animal should not be immobilized but should be able to move. Many behavioral changes occur during long time scales and require recording over many hours of behavior. This also makes it necessary to culture the worms in the presence of food. How can worms be cultured and their neural activity imaged over long time scales? Agarose Microchamber Imaging (AMI) was previously developed to culture and observe small larvae and has now been adapted to study all life stages from early L1 until the adult stage of C. elegans. AMI can be performed on various life stages of C. elegans. Longterm calcium imaging is achieved without immobilizing the animals by using short externally triggered exposures combined with an electron multiplying charge-coupled device (EMCCD) camera recording. Zooming out or scanning can scale up this method to image up to 40 worms in parallel. Thus, a method is described to image behavior and neural activity over long time scales in all life stages of C. elegans.

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Non-microfluidic methods for imaging live C. elegans

Cited by 15 publications

References 13 publications

Stress and timing associated with Caenorhabditis elegans immobilization methods

Stress and timing associated with Caenorhabditis elegans immobilization methods

Caenorhabditis elegans as a Model Host to Monitor the Candida Infection Processes

Agarose Microchambers for Long-term Calcium Imaging of <em>Caenorhabditis elegans</em>

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