Fluorescence images were acquired essentially as described previously (Aoki & Matsuda, 2009 (link)). Briefly, cells cultured on glass-base dishes were observed under an IX83 inverted microscope (Olympus) equipped with a UPlanFL-PH 10×/0.3 (Olympus), a UPlanSApo 20×/0.75 (Olympus), or a UPlanSApo 40×/0.95 objective lens (Olympus), a DOC CAM-HR CCD camera (Molecular Devices), a Spectra-X0 light engine (Lumencor Inc.), an IX3-ZDC laser-based autofocusing system (Olympus), an electric XY stage (Sigma Koki) and a stage top incubator (Tokai Hit). The filters and dichromatic mirrors used for time-lapse imaging were as follows: for FRET imaging, a 438/24 excitation filter incorporated in the Spectra-X light engine, an FF458-Di02-25x36 dichromatic mirror (Semrock), and FF01-483/32-25 and FF01-542/27-25 emission filters (Semrock) for CFP and FRET, respectively.
Ix3 zdc laser based autofocusing system
Manufactured by Olympus
Sourced in Japan
The IX3-ZDC is a laser-based autofocusing system developed by Olympus. Its core function is to automatically maintain the optimal focus during microscope observations.
Automatically generated - may contain errors
Lab products found in correlation
Ix83 inverted microscope, by Olympus (2 mentions)
Doc cam hr ccd camera, by Molecular Devices (2 mentions)
Spectra x light engine, by IDEX Corporation (2 mentions)
Ff01 483 32 25, by IDEX Corporation (2 mentions)
Stage top incubator, by Tokai Hit (2 mentions)
Uplansapo 20 0, by Olympus (1 mentions)
Ff458 di02 25x36, by IDEX Corporation (1 mentions)
Uplansapo 20x 0, by Olympus (1 mentions)
Ff01 542 27 25, by IDEX Corporation (1 mentions)
2 protocols using ix3 zdc laser based autofocusing system
1
Fluorescence Imaging Microscopy Protocol
2
FRET Imaging Protocols for Live Cell Analysis
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FRET images were obtained and processed under essentially the same conditions and procedures as previously described (Aoki and Matsuda, 2009) . Briefly, cells were imaged with an IX83 inverted microscope (Olympus, Tokyo, Japan) equipped with a UPlanFL-PH 10x/0.3 (Olympus), a UPlanSApo 20x/0.75 (Olympus), or a UPlanSApo 40x/0.95 objective lens (Olympus), a DOC CAM-HR CCD camera (Molecular Devices, Sunnyvale, CA), a Spectra-X light engine (Lumencor Inc., Beaverton, OR), an IX3-ZDC laser-based autofocusing system (Olympus), an electric XY stage (Sigma Koki, Tokyo, Japan), and a stage top incubator (Tokai Hit, Fujinomiya, Japan). The filters and dichromatic mirrors used for time-lapse imaging were as follows: for FRET imaging, a 438/24 excitation filter (incorporated in the Spectra-X light engine), a FF458-Di02-25x36 (Semrock, Rochester, NY) dichromatic mirror, and FF01-483/32-25 (Semrock) and FF01-542/27-25 (Semrock) emission filter for CFP and FRET, respectively. For mCherry and mScarlet-I imaging, a 575/25 excitation filter, a glass dichromatic mirror (Olympus), and FF01-624/40-25 (Semrock) emission filters were used.
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