Cells were imaged using a Nikon TE-2000E inverted microscope with a 603 1.49 NA TIRF objective, Andor Revolution XD spinning disk confocal system, and 488 and 568 nm solid-state lasers. Cells were imaged in Opti-MEM or Leibowitz's L15 medium (Gibco), 5% FBS, at 37°C. Time lapses were acquired using an Andor iXon+ EM-CCD camera using Andor IQ. Original 16-bit tif files acquired directly from the camera were used for image analysis.
Te 2000e inverted microscope
Manufactured by Oxford Instruments
The TE-2000E is an inverted microscope designed for laboratory use. It features a compact and modular design, suitable for a variety of applications. The core function of the TE-2000E is to provide high-quality optical imaging and observation of samples.
Automatically generated - may contain errors
3 protocols using te 2000e inverted microscope
1
Live-cell Imaging of Cellular Dynamics
2
Live-cell Imaging of Cellular Dynamics
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Cells were imaged using a Nikon TE-2000E inverted microscope with a 603 1.49 NA TIRF objective, Andor Revolution XD spinning disk confocal system, and 488 and 568 nm solid-state lasers. Cells were imaged in Opti-MEM or Leibowitz's L15 medium (Gibco), 5% FBS, at 37°C. Time lapses were acquired using an Andor iXon+ EM-CCD camera using Andor IQ. Original 16-bit tif files acquired directly from the camera were used for image analysis.
3
Imaging Division Plane Dynamics in Embryos
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One-cell stage embryos were mounted in microdevices as for division plane imaging and four-cell stage embryos were mounted on slides with 2% agarose pads. Embryos were imaged on a Nikon TE2000-E inverted microscope equipped with a 60 × 1.40 NA objective, an EM-CCD camera (iXon; Andor Technology; EM-Gain = 220, Exposure = 100 ms), and a krypton-argon 2.5 W water-cooled laser. For one-cell stage embryos, division planes were reconstructed from 30 × 1 µm stacks acquired every 20 s with 20% laser power and photo-bleaching was performed by 2 sweeps of a 488 nm laser with 100% power and 500 µs dwell time. For four-cell stage embryos, division planes were reconstructed from 16 × 1 µm stacks acquired every 10 s with 50% laser power and photo-bleaching was performed by 2 sweeps of a 488 nm laser with 100% power and 100 µs dwell time. For four-cell stage embryos, the time between the prebleached and first postbleached images was 6 s.
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