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Axiovert s100 microscope

Manufactured by Eppendorf
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Sourced in United Kingdom

The Axiovert S100 is an inverted microscope designed for cell culture observation and analysis. It features a stable and vibration-resistant body, a long working distance objective, and can accommodate a range of accessories to support various microscopy techniques. This product is suitable for routine cell culture monitoring and imaging.

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Lab products found in correlation

Spin x filter, by Corning (3 mentions) Femtotip, by Eppendorf (3 mentions) Ix70 imaging system, by Olympus (2 mentions) Femtojet microinjector, by Eppendorf (2 mentions)

3 protocols using axiovert s100 microscope

1

Microinjection and Axonal Transport Imaging

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Dissociated SCG neurons were microinjected using a Zeiss Axiovert S100 microscope with an Eppendorf FemtoJet microinjector and Eppendorf TransferMan® micromanipulator. Plasmids were diluted in 0.5× PBS (without CaCl2 and MgCl2) and filtered using a Spin-X filter (Costar). The mix was injected directly into the nuclei of SCG neurons using Eppendorf Femtotips. Approximately 100 neurons were injected per dish. Injected plasmids were allowed to express for 16 h before CCCP treatment. Plasmids were injected at the following concentrations: 30 ng/μl NMNAT2-EGFP, 30 ng/μl pDsRed2-N1. Time-lapse imaging of axonal transport was performed on an Olympus IX70 imaging system with 100×/1.35 Oil objective. During imaging, cell cultures were maintained at 37 °C and 5% CO2 in an environment chamber. Images were captured at 4 frames per sec for 2 min. Three neurites per condition were imaged in every individual experiment.
Loreto A., Hill C.S., Hewitt V.L., Orsomando G., Angeletti C., Gilley J., Lucci C., Sanchez-Martinez A., Whitworth A.J., Conforti L., Dajas-Bailador F, & Colemana M.P. (2019). Mitochondrial impairment activates the Wallerian pathway through depletion of NMNAT2 leading to SARM1-dependent axon degeneration. Neurobiology of disease, 134, 104678.
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2

Microinjection of SCG Neurons

Cited 1 time
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Microinjection was performed as previously described16 (link) with minor modifications. Dissociated cells were microinjected using a Zeiss Axiovert S100 microscope (Cambridge, UK) with an Eppendorf FemtoJet transjector and 5171 micromanipulator system and Eppendorf Femtotips (Stevenage, UK). Plasmids were diluted in 0.5 × PBS to a concentration of 100 ng/μl and passed through a Spin-X filter (Costar, Fisher, Loughborough, UK). The mix was injected directly into the nuclei of SCG neurons in dissociated cultures. All plasmids were co-injected with DsRed2 at a concentration of 25 ng/μl. In all, 70–200 neurons were injected per dish. Injection of relatively few neurons per dish facilitated visualization of individual labelled neurites, as neurites tend to cluster together in bundles.
Di Stefano M., Nascimento-Ferreira I., Orsomando G., Mori V., Gilley J., Brown R., Janeckova L., Vargas M.E., Worrell L.A., Loreto A., Tickle J., Patrick J., Webster J.R., Marangoni M., Carpi F.M., Pucciarelli S., Rossi F., Meng W., Sagasti A., Ribchester R.R., Magni G., Coleman M.P, & Conforti L. (2014). A rise in NAD precursor nicotinamide mononucleotide (NMN) after injury promotes axon degeneration. Cell Death and Differentiation, 22(5), 731-742.
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3

Microinjection and Axonal Transport Imaging

Check if the same lab product or an alternative is used in the 5 most similar protocols
Dissociated SCG neurons were microinjected using a Zeiss Axiovert S100 microscope with an Eppendorf FemtoJet microinjector and Eppendorf TransferMan® micromanipulator. Plasmids were diluted in 0.5× PBS (without CaCl2 and MgCl2) and filtered using a Spin-X filter (Costar). The mix was injected directly into the nuclei of SCG neurons using Eppendorf Femtotips. Approximately 100 neurons were injected per dish. Injected plasmids were allowed to express for 16 h before CCCP treatment. Plasmids were injected at the following concentrations: 30 ng/μl NMNAT2-EGFP, 30 ng/μl pDsRed2-N1. Time-lapse imaging of axonal transport was performed on an Olympus IX70 imaging system with 100×/1.35 Oil objective. During imaging, cell cultures were maintained at 37 °C and 5% CO2 in an environment chamber. Images were captured at 4 frames per sec for 2 min. Three neurites per condition were imaged in every individual experiment.
Loreto A., Hill C.S., Hewitt V.L., Orsomando G., Angeletti C., Gilley J., Lucci C., Sanchez-Martinez A., Whitworth A.J., Conforti L., Dajas-Bailador F, & Colemana M.P. (2019). Mitochondrial impairment activates the Wallerian pathway through depletion of NMNAT2 leading to SARM1-dependent axon degeneration. Neurobiology of disease, 134, 104678.
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