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Cytotune emgfp sendai fluorescence reporter

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The CytoTune emGFP Sendai fluorescence reporter is a laboratory instrument designed to visualize and track the expression of enhanced green fluorescent protein (emGFP) in cells. It utilizes Sendai virus technology to deliver the emGFP gene into target cells, enabling the monitoring of cellular processes through fluorescence microscopy.

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Lab products found in correlation

Evos fl inverted microscope, by Thermo Fisher Scientific (2 mentions) Alexafluor 647 conjugate ctb, by Thermo Fisher Scientific (2 mentions) Dii crystal, by Thermo Fisher Scientific (2 mentions) Lentibrite rfp control lentiviral biosensor, by Merck Group (2 mentions) Fast green fcf, by Merck Group (2 mentions)

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CytoTune (22 citations)

5 protocols using cytotune emgfp sendai fluorescence reporter

1

Multimodal Labeling of Organoids

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ALI-COs were visualised on an EVOS FL inverted microscope (ThermoFisher) and using a microinjection capillary <0.2 μl of 1 mg/ml AlexaFluor 647-conjugate CTB (ThermoFisher, C34778) or 1-4 DiI crystals (ThermoFisher, D3911) were applied to the target region. DiI tracing was performed multiple times but due to difficulty with dye uptake and low signal-to-noise ratio only in one experiment was tracing of sufficient quality for analyis. Similarly, to achieve sparse neuronal labeling, ALI-COs were injected with <0.2 μl of CytoTune emGFP Sendai fluorescence reporter (ThermoFisher, A16519). Four days after CTB and DiI labeling and 5 days after viral emGFP labelling the samples were fixed for histological analyses.
Giandomenico S.L., Mierau S.B., Gibbons G.M., Wenger L.M., Masullo L., Sit T., Sutcliffe M., Boulanger J., Tripodi M., Derivery E., Paulsen O., Lakatos A, & Lancaster M.A. (2019). Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output. Nature neuroscience, 22(4), 669-679.
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2

Labeling and Clonal Analysis of Organoids

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Organoids were labelled using a CytoTune™ EmGFP Sendai Fluorescence Reporter (ThermoFisher, A16519). The stock solution of the virus was diluted with PBS to 1:10 and kept on ice. The solution was visualized with Fast Green FCF (Sigma, F7252) and was delivered to the ventricles at 45d using a pulled capillary. Organoids were immediately subjected to hormone treatment. The media was changed every day. Organoids were collected for analysis at 6- and 8-days post-labelling. At 8 days post labelling, some of the organoids were injected with LentiBrite™ RFP Control Lentiviral Biosensor (Millipore, 17-10409), using the same procedure as with EmGFP Sendai virus. These organoids were kept in IDM+A with dissolved Matrigel® for a further 8 days and then fixed for clonal analysis.
Kelava I., Chiaradia I., Pellegrini L., Kalinka A.T, & Lancaster M.A. (2022). Androgens increase excitatory neurogenic potential in human brain organoids. Nature, 602(7895), 112-116.
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3

Multimodal Labeling of Organoids

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ALI-COs were visualised on an EVOS FL inverted microscope (ThermoFisher) and using a microinjection capillary <0.2 μl of 1 mg/ml AlexaFluor 647-conjugate CTB (ThermoFisher, C34778) or 1-4 DiI crystals (ThermoFisher, D3911) were applied to the target region. DiI tracing was performed multiple times but due to difficulty with dye uptake and low signal-to-noise ratio only in one experiment was tracing of sufficient quality for analyis. Similarly, to achieve sparse neuronal labeling, ALI-COs were injected with <0.2 μl of CytoTune emGFP Sendai fluorescence reporter (ThermoFisher, A16519). Four days after CTB and DiI labeling and 5 days after viral emGFP labelling the samples were fixed for histological analyses.
Giandomenico S.L., Mierau S.B., Gibbons G.M., Wenger L.M., Masullo L., Sit T., Sutcliffe M., Boulanger J., Tripodi M., Derivery E., Paulsen O., Lakatos A, & Lancaster M.A. (2019). Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output. Nature neuroscience, 22(4), 669-679.
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4

Visualizing Axons and Cytoskeleton in Neuronal Cultures

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Dissociated neuronal cultures used for immunofluorescence staining of ribosomal subunits were fed with 200 µl of SFSC medium supplemented with CytoTune EmGFP Sendai Fluorescence Reporter (Thermo Fisher Scientific, A16519, 8.1 × 107 CIU/ml) diluted 1:200. After 3–4 days, the cells started displaying EmGFP signal. Approximately 2 weeks after dissociation, cultures produced thin EmGFP+ axons and the cultures were fixed for analysis. SiR-tubulin (Lukinavičius et al., 2014 (link)) was reconstituted in sterile DMSO to a concentration of 1 mM. For staining of ALI-COs SiR-tubulin was diluted to a final concentration of 1 µM in SFSC medium and applied dropwise to the top of the slice using a controlled oral-suction pipetting apparatus and care was taken not to disturb the grids. After approximately 1 hr at 37 °C and 5 % CO2 samples were ready for imaging.
Hoffmann P.C., Giandomenico S.L., Ganeva I., Wozny M.R., Sutcliffe M., Lancaster M.A, & Kukulski W. (2021). Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids. eLife, 10, e70269.
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5

Organoid Labeling and Analysis

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Organoids were labelled using a CytoTune™ EmGFP Sendai Fluorescence Reporter (ThermoFisher, A16519). The stock solution of the virus was diluted with PBS to 1:10 and kept on ice. The solution was visualized with Fast Green FCF (Sigma, F7252) and was delivered to the ventricles at 45d using a pulled capillary. Organoids were immediately subjected to hormone treatment. The media was changed every day. Organoids were collected for analysis at 6-and 8days post-labelling. At 8 days post labelling, some of the organoids were injected with LentiBrite™ RFP Control Lentiviral Biosensor (Millipore, 17-10409), using the same procedure as with EmGFP Sendai virus. These organoids were kept in IDM+A with dissolved Matrigel® for a further 8 days and then fixed for clonal analysis.
Kelava I., Chiaradia I., Pellegrini L., Kalinka A.T., & Lancaster M.A. (2020). Male sex hormones increase excitatory neuron production in developing human neocortex.
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