Cells were imaged in 96-well glass-bottom microplates (Greiner Bio-One) containing indicated media with an inverted microscope (Ti-E; Nikon) using a Plan Apochromat IR 60× 1.27 NA objective (Nikon) and Spectra X LED light source (Lumencor) at room temperature. 3D light microscopy data were collected using the triggered Z-Piezo system (Nikon) and orca flash 4.0 camera (Hamamatsu). 3D data were processed using NIS-elements AR (Nikon), Huygens Professional 16.10 (Scientific Volume Imaging), ImageJ v.2.0.0., and Photoshop (Adobe Systems) software.
Z piezo system
Manufactured by Nikon
The Z-Piezo system is a high-precision vertical positioning stage designed for Nikon's microscopy and imaging equipment. It utilizes piezoelectric actuators to provide smooth, accurate, and repeatable control of the Z-axis positioning with sub-micron resolution. The core function of the Z-Piezo system is to enable precise, automated focusing and positioning of samples during microscopy and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Spectrax led light source, by Lumencor (3 mentions)
Orca flash 4.0 camera, by Hamamatsu Photonics (3 mentions)
Plan apochromat ir 60 1.27 na objective, by Nikon (2 mentions)
96 well glass bottom microplates, by Greiner (2 mentions)
Nis elements ar, by Nikon (2 mentions)
Huygens professional 16, by Scientific Volume Imaging (1 mentions)
Zyla 4.2 plus scmos camera, by Oxford Instruments (1 mentions)
3 protocols using z piezo system
1
3D Live-Cell Microscopy Imaging
2
High-resolution 3D microscopy of cells
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Cells were viewed in 96-well microplates with a glass bottom (Greiner bio-one) containing indicated growth or starvation media at RT with an inverted microscope (Ti-E; Nikon) using a Plan Apochromat IR 60× 1.27 NA objective (Nikon), Spectra X LED light source (Lumencor), and acquisition software NIS elements AR (Nikon). 3D light microscopy data were collected using the triggered Z-Piezo system (Nikon) and orca flash 4.0 camera (Hamamatsu). 3D data were processed using NIS-elements AR; deconvolution of imaging data was performed with 3D deconvolution 3D-Fast in NIS-elements AR (Figs. 2 E , 3 C , 4 A , and 5 A ) or Huygens Professional 15.10 (Scientific Volume Imaging; Figs. 1, B and C ; Fig. 2 C ; Fig. 4 D ; and Fig. S1, A, C, and G). Photoshop (Adobe) software was used to make linear adjustments in brightness.
3
High-Resolution Microscopy of Cellular Dynamics
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Cells were transferred to 96-well glass-bottom microplates (Greiner Bio-One) containing indicated media. For fixed timepoint analyses, cells were imaged at room temperature with an inverted microscope (Nikon Ti-E) using a Plan Apochromat IR 60× 1.27 numerical aperture objective (Nikon) and Spectra X LED light source (Lumencor). Three-dimensional light microscopy data were collected using the triggered Z-Piezo system (Nikon) and orca flash 4.0 camera (Hamamatsu). High-speed confocal imaging was performed with a Dragonfly 500 series spinning disk microscope (Andor, Oxford Instruments) equipped with a Zyla 4.2 Plus sCMos camera (Andor) using a Lamba CFI-Plan Apochromat 60× 1.4 numerical aperture oil immersion objective (Nikon). Three-dimensional data were processed using Fusion software (Andor), and Fiji ImageJ Version 2.1.0. Deconvolution was performed using Huygens Professional 16.10 where indicated. Data are shown as single sections if not stated otherwise.
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