Experiments were performed on a Dmi8 upright microscope (Leica, Wetzlar, Germany). Images were acquired with an ORCA-Flash4.0 C13440-20C camera (Hamamatsu photonics, Hamamatsu, Japan) controlled by LAS X software (Leica). For determination of membrane localization of GFP–tubbyCT constructs, CHO cells were co-transfected with a membrane marker (Lyn11–RFP) or catalytically inactive Ci-VSP (RFP-Ci-VSP C363S). Line profiles across the cells were analyzed. GFP fluorescence intensities at membrane marker peaks were averaged and normalized to average cytosolic fluorescence.
Orca flash4.0 c13440 20c camera
Manufactured by Hamamatsu Photonics
Sourced in Japan
The ORCA-Flash4.0 C13440-20C is a scientific-grade CMOS camera manufactured by Hamamatsu Photonics. It features a 4-megapixel sensor with a pixel size of 6.5 µm and a maximum frame rate of 100 frames per second. The camera is capable of high-sensitivity and low-noise imaging, making it suitable for a variety of scientific and industrial applications.
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Lab products found in correlation
3 protocols using orca flash4.0 c13440 20c camera
1
Membrane Localization of GFP-tubbyCT Constructs
2
TIRF Microscopy of Tubbyclusters
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TIRF imaging was done on a Dmi8 upright microscope (Leica, Wetzlar, Germany) equipped with an Infinity TIRF module (Leica), a HC PL APO 100×/1.47 oil objective (Leica), and a widefield laser (Leica). GFP fluorescence was excited at 488 nm and imaged through a GFP-T (505 to 555 nm) emission filter (Leica). Images were acquired every 6 s with an ORCA-Flash4.0 C13440-20C camera (Hamamatsu photonics, Hamamatsu, Japan) controlled by LAS X software (Leica). During imaging, cells were perfused with an extracellular solution [5.8 mM KCl, 144 mM NaCl, 0.9 mM MgCl2, 1.3 mM CaCl2, 0.7 mM NaH2PO4, 5.6 mM d -glucose, and 10 mM Hepes (pH 7.4)]. Data were analyzed by ImageJ software, and tubbyCT clusters were detected by an algorithm described previously (25 (link)).
3
Membrane Localization of GFP-tubbyCT
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Experiments were performed on a Dmi8 upright microscope (Leica, Wetzlar, Germany).
Images were acquired with an ORCA-Flash4.0 C13440-20C camera (Hamamatsu photonics, Hamamatsu, Japan) controlled by LAS X software (Leica). For determination of membrane localisation of GFP-tubbyCT constructs, CHO cells were co-transfected with a catalytically inactive Ci-VSP (RFP-Ci-VSP C363S) as membrane marker. Line profiles across the cells were analysed. GFP fluorescence intensities at Ci-VSP C363S membrane peaks were averaged and normalized to average cytosolic fluorescence.
Images were acquired with an ORCA-Flash4.0 C13440-20C camera (Hamamatsu photonics, Hamamatsu, Japan) controlled by LAS X software (Leica). For determination of membrane localisation of GFP-tubbyCT constructs, CHO cells were co-transfected with a catalytically inactive Ci-VSP (RFP-Ci-VSP C363S) as membrane marker. Line profiles across the cells were analysed. GFP fluorescence intensities at Ci-VSP C363S membrane peaks were averaged and normalized to average cytosolic fluorescence.
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