About 24–36 h after transfection, coverslides were transferred into a measuring chamber and rinsed once with a buffer containing 148 mM NaCl, 5.4 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 10 mM HEPES, pH 7.3. The measuring chamber was then transferred onto a ZEISS AxioObserver A1 epifluorescence microscope equipped with an oil immersion Plan Apochromat 63× objective, polychrome V light source (TILL Photonics, Gräfelfing, Germany), DV2 QuadlView (Photometrics, Tucson, AZ, USA), and a Cool-SNAP-HQ CCD-camera (Visitron Systems, Puchheim, Germany). Cells were then treated with 10 μM forskolin added in the same buffer (all chemicals were purchased from Sigma-Aldrich). To monitor FRET between GFP and mCherry, cells were excited at 490 nm, the emission light was split into two channels using the DV2 QuadView (565dcxr dichroic mirror) and detected at 515 ± 15 (GFP) and 590 ± 20 (mCherry) nm. FRET changes were monitored using VISIVIEW software (Visitron Systems) as the emission ratio of mCherry over GFP and analyzed as previously described [39 (link)].
Cool snap hq ccd camera
Manufactured by Visitron
Sourced in United States
The Cool-SNAP-HQ CCD-camera is a high-quality imaging device designed for scientific and industrial applications. It features a charge-coupled device (CCD) sensor and advanced cooling technology to capture high-resolution images with low noise and high sensitivity.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using cool snap hq ccd camera
1
FRET Measurement of Protein Interaction
2
FRET Imaging of Thyroid Follicles
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Glass-bottom Petri dishes containing thyroid follicles isolated from CAG-Epac1-camps mice were placed on a Zeiss Axiovert 200 inverted microscope equipped with an oil-immersion 63 × objective, a polychrome IV light source (Till Photonics), a 505 DCXR beam splitter, and a Cool SNAP-HQCCD-camera (Visitron Systems). Forster resonance energy transfer (FRET) was monitored using MetaFluor 5.0 software (Molecular Devices) as the ratio between emission at 535 ± 20 nm (yellow fluorescent protein [YFP]) and emission at 480 ± 15 nm (cyan fluorescent protein [CFP]), upon excitation at 436 ± 10 nm. The imaging data were analyzed using MetaMorph 5.0 (Molecular Devices) and Prism (GraphPadSoftware) software, by correcting for spillover of CFP into the 535-nm channel and direct YFP excitation, to give corrected YFP/CFP ratio data. Images were acquired every 5 seconds, with 5 ms of illumination time. Photo bleaching was negligible during a 30-minute observation. Experiments were done at 37 °C. Thyroid follicles were kept under laminar-flow perfusion with a custom-built apparatus that allows for the rapid exchange between different extracellular solutions.
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