Author response: A tunable refractive index matching medium for live imaging cells, tissues and model organisms

Boothe, Tobias; Hilbert, Lennart; Heide, Michael; Berninger, Lea; Huttner, Wieland B.; Zaburdaev, Vasily; Vastenhouw, Nadine L.; Myers, Eugene W.; Drechsel, David; Rink, Jochen C.

doi:10.7554/elife.27240.022

Cited by 3 publications

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Linalool acts as a fast and reversible anesthetic inHydra

Goel

Wang

Martin

et al. 2019

Preprint

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15The ability to make transgenic Hydra lines has opened the door for quantitative in vivo studies of 16 Hydra regeneration and physiology. These studies commonly include excision, grafting and 17 transplantation experiments along with high-resolution imaging of live animals, which can be 18 challenging due to the animal's response to touch and light stimuli. While various anesthetics 19 have been used in Hydra studies over the years, they tend to be toxic over the course of a few 20 hours or their long-term effects on animal health have not been studied. Here we show that the 21 monoterpenoid linalool is a useful anesthetic for Hydra. Linalool is easy to use, non-toxic, fast 22 acting, and reversible. It has no detectable long-term effects on cell viability or cell proliferation. 23 We demonstrate that the same animal can be immobilized in linalool multiple times at intervals 24 of several hours for repeated imaging over 2-3 days. This uniquely allows for in vivo imaging of 25 dynamic processes such as head regeneration. We further directly compare linalool to currently 26 used anesthetics and show its superior performance. Because linalool, which is frequently 27 utilized in perfumes and cosmetic products, is also non-hazardous to humans, it will be a useful 28 tool for Hydra research in both research and teaching contexts. 29 30 31 112

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Linalool acts as a fast and reversible anesthetic inHydra

Goel

Wang

Martin

et al. 2019

Preprint

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Processive activity of the replicative DNA polymerases in the replisome of live eukaryotic cells

Kapadia

El-Hajj

Zheng

et al. 2019

Preprint

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DNA replication is carried out by a multi-protein machine called the replisome. In Saccharomyces cerevisiae, the replisome is composed of over 30 different proteins arranged into multiple subassemblies, each performing distinct activities. Synchrony of these activities is required for efficient replication and preservation of genomic integrity. How this is achieved is particularly puzzling at the lagging strand, where current models of the replisome architecture propose turnover of the canonical lagging strand polymerase, Pol δ, at every cycle of Okazaki fragment synthesis. Here we established single-molecule fluorescence microscopy protocols to study the binding kinetics of individual replisome subunits in live S. cerevisiae. Our results show long residence times for most subunits at the active replisome, supporting a model where all subassemblies bind tightly and work in a coordinated manner for extended periods, including Pol δ, hence redefining the architecture of the active eukaryotic replisome.

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3D imaging shows nano- and microparticles are internalized by salmon skin and corneal epithelial cells

Thiyagarajan,

Stette,

Afzal

et al. 2024

Front. Mar. Sci.

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The rising problem of plastic pollution is becoming one of the major environmental issues for the world. In the ocean, plastics undergo degradation into smaller microplastics (MPs) and nanoplastics (NPs). Wild fish and farmed salmon would likely be exposed to these NPs and MPs both through skin and through skin wounds. Keratocyte cells, located in the skin epithelial layer, are scavenger cells which may remove foreign materials and maintain the salmon’s health. They are therefore first in line to handle and to suffer from MP and NP exposure. While the impacts of MPs have been well studied in many different organisms, much less is known about the effects of NP exposure, particularly at the subcellular level. Here, we have used holotomographic and fluorescence microscopy to show that both skin and corneal salmon keratocyte cells fully internalize 500–1000 nm polystyrene particles, as well as inorganic 500 nm silica particles. The fact that corneal epithelial cells also take up particles is novel. Furthermore, some of these particles likely end up in lysosomal compartments within 2 hours of exposure. Here, we show that both conventional and new modalities of microscopy have a role to play to understand how micro- and nano particles affect epithelial cells.

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Author response: A tunable refractive index matching medium for live imaging cells, tissues and model organisms

Cited by 3 publications

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Linalool acts as a fast and reversible anesthetic inHydra

Linalool acts as a fast and reversible anesthetic inHydra

Processive activity of the replicative DNA polymerases in the replisome of live eukaryotic cells

3D imaging shows nano- and microparticles are internalized by salmon skin and corneal epithelial cells

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