Confocal Imaging of Cy3-Labeled Cells

As previously detailed [28 (link), 31 (link)], confocal images were acquired in a FV1000 laser scanning microscope (Olympus) using an Olympus UPlanSApo 60x oil immersion objective (NA = 1.35). Cy3 were excited using a He-Ne green laser at 543 nm (average power at the sample, 700 nW). Fluorescence was detected with a photomultiplier set in the pseudo photon-counting detection mode, using 560–660 nm filtering for Cy3 detection. An averaged image was obtained from 10 consecutive images acquired per cell.

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Francia M., Stortz M., Echegaray C.V., Oses C., Verneri P., Petrone M.V., Toro A., Waisman A., Miriuka S., Cosentino M.S., Levi V, & Guberman A. (2021). SUMO conjugation susceptibility of Akt/protein kinase B affects the expression of the pluripotency transcription factor Nanog in embryonic stem cells. PLoS ONE, 16(7), e0254447.

Publication 2021

FV1000 laser scanning microscope UPlanSApo 60x oil

Cell Fluorescence Immersion Laser scanning microscope

Corresponding Organization : Consejo Nacional de Investigaciones Científicas y Técnicas

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Variable analysis

independent variables

Laser power (700 nW)

dependent variables

Fluorescence intensity

control variables

Confocal microscope (FV1000)
Olympus UPlanSApo 60x oil immersion objective (NA = 1.35)
Cy3 fluorophore
He-Ne green laser at 543 nm
Photomultiplier in pseudo photon-counting detection mode
560–660 nm filtering for Cy3 detection
Averaged image from 10 consecutive images per cell

controls

Positive control: Not specified
Negative control: Not specified

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