Imaging was performed at room temperature using the inverted Nikon Ti Eclipse spinning disk confocal microscope with a 100× NA 1.4 oil immersion objective and a 2× Andor Ultra 888 EMCCD camera (BioFrontiers Advanced Light Microscopy Core) (Figs. 1 , S1 , S2 , S3 , and 4 ), the Nikon Spinning Disk Super Resolution by Optical Pixel Reassignment (SoRa) microscope with a 20× NA 0.75 air (Fig. S2 H ), 100× NA 1.45 oil immersion objective, and a Hamamatsu ORCA Fusion BT sCMOS Camera using 1× (Figs. 3 , 4 , S4 , 5 , S5 , and 6 ) or 2.8× magnification (Fig. 4 F ). Nikon NIS-Elements was used as acquisition software. Every experiment was performed in biological replicates/different days and indicated in the figures. More than three images were recorded at room temperature per biological replicate. The number of analyzed cells is indicated in the figures.
Orca fusion bt scmos camera
Manufactured by Hamamatsu Photonics
The Orca-Fusion BT sCMOS cameras from Hamamatsu Photonics are designed to provide high-performance imaging capabilities. They feature a back-thinned sCMOS sensor with a large active area and high quantum efficiency. The cameras offer a high frame rate, low noise, and wide dynamic range, making them suitable for a variety of scientific and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ti2, by Nikon (3 mentions)
Mini scanner module, by Bruker (3 mentions)
Structured illumination microscope n sim, by Oxford Instruments (3 mentions)
Ultra du 897 emccd camera, by Oxford Instruments (3 mentions)
Prime 95b scmos, by Hamamatsu Photonics (3 mentions)
A1 microscope, by Nikon (3 mentions)
Eclipse ti, by Nikon (1 mentions)
Sr apo tirf 100x 1.49 na, by Oxford Instruments (1 mentions)
5 protocols using orca fusion bt scmos camera
1
Confocal Microscopy Imaging Protocol
2
Cerebellar Vermis Immunolabeling Protocol
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To minimize any effects of tissue laterality, all analyses were performed using sections obtained from the cerebellar vermis (between 0 and 1800 μm from the cerebellar midline). Sections for immunolabeling were selected at random from within this range to avoid any systematic bias. Sampling was performed in linear regions of lobules 5, 8, and 10 as described above (Fig. 1). Images were acquired from well-labeled sections using a Nikon W1-CSU Dual Spinning Disk Confocal microscope fitted with a Hamamatsu Orca Fusion BT sCMOS camera. Images of immunolabeling were collected as Z-stacks of 12 μm total depth at 0.3 μm step size, using a 60x objective lens with 1.4 numerical aperture. Image resolution was 0.10833 μm/px.
3
Advanced Microscopy Techniques for Live Imaging
4
Advanced Microscopy Techniques for Live Imaging
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Laser scanning confocal imaging was conducted using Nikon A1 Microscope equipped with 405, 488, 561 and 645 nm LASERs, Apo TIRF 100×/1.45 NA, Plan Apo 60×/1.4 NA, Plan Fluor ELWD 40×/0.6 NA objectives. Live-cell imaging was conducted using a Nikon Ti2 Eclipse equipped with 488, 561 and 645 nm excitation LASERs, Apo TIRF 100×/1.49 NA and Plan Fluor 40×/1.3 NA objectives, a Hamamatsu X1 spinning disk, and Photometrics Prime 95B sCMOS or Hamamatsu Orca-Fusion BT sCMOS cameras. FRAP was also conducted using a Bruker mini-scanner module capable of producing ROI specific 405 nm photo-stimulation. Images were deconvolved and/or denoised using Nikon Elements software. Super-resolution imaging was performed using a Nikon Structured Illumination Microscope (N-SIM) equipped with 405, 488, 561 and 640 nm LASERs, an SR Apo TIRF 100×/1.49 NA objective, and an Andor iXon Ultra DU-897 EMCCD camera. Images were reconstructed using Nikon Elements software. For imaging in all microscope modalities, gain was matched between samples during image acquisition.
5
Multimodal Microscopy Imaging Protocol
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Laser scanning confocal imaging was conducted using Nikon A1 Microscope equipped with 405, 488, 561 and 645 nm LASERs, Apo TIRF 100x/1.45 NA, Plan Apo 60x/1.4 NA, Plan Fluor ELWD 40x/0.6 NA objectives. Live-cell imaging was conducted using a Nikon Ti2 Eclipse equipped with 488, 561 and 645 nm excitation LASERs, Apo TIRF 100x/1.49 NA and Plan Fluor 40x/1.3 NA objectives, a Hamamatsu X1 spinning disk, and Photometrics Prime 95B sCMOS or Hamamatsu Orca-Fusion BT sCMOS cameras. FRAP was also conducted using a Bruker mini-scanner module capable of producing ROI specific 405 nm photo-stimulation. Images were deconvolved and/or denoised using Nikon Elements software. Super-resolution imaging was performed using a Nikon Structured Illumination Microscope (N-SIM) equipped with 405, 488, 561 and 640 nm LASERs, an SR Apo TIRF 100x/1.49 NA objective, and an Andor iXon Ultra DU-897 EMCCD camera. Images were reconstructed using Nikon Elements software. For imaging in all microscope modalities, gain was matched between samples during image acquisition.
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