The immunostained samples were imaged using an inverted Nikon-Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU-888 camera (Oxford Instruments, United Kingdom) and a pE-100 LED illumination system (CoolLED Ltd, Andover, United Kingdom). Fluorescent images of immunostained HUVECs on the deformable PDMS membranes were acquired with a 20X, 0.75 NA air objective (Plan Apo, Nikon, Japan), using FITC, TRITC, and DAPI filters, respectively. Large images up to 6 mm × 6 mm were acquired by using the Custom Multipoint/Large Image function of NIS Elements (NIS Elements, Nikon, Japan) in combination with an autofocus routine followed by automated stitching.
Andor du 888 camera
Manufactured by Oxford Instruments
Sourced in Japan
The Andor DU-888 camera is a scientific-grade, back-illuminated, deep-cooled CCD camera. It features high quantum efficiency, low noise, and high readout speed, making it suitable for a range of scientific applications that require high-performance imaging.
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Lab products found in correlation
5 protocols using andor du 888 camera
1
Imaging Deformable HUVEC Membranes
2
Fluorescent Cell Cycle Imaging
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Cell cycle movies were acquired using an inverted Nikon-Ti wide-field microscope (Nikon, Japan) equipped with an Orca R-2 CCD camera (Hamamatsu Photonics, Japan) or a Nikon-Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU-888 camera (Oxford Instruments, United Kingdom), both with an incubation chamber (Life Imaging Services, Switzerland) to control temperature, CO2, and humidity. Images were collected using a 40× objective (Plan Fluor 40× Oil DIC H N2). Multiple nonoverlapping fields capturing all positions with quantum dots were recorded in parallel (ΔT = 1 h, total duration ∼30 h for HeLa Fucci and ∼48 h for MCF7).
At each time of measurement, a transmission and two fluorescent images of the nuclei of the cells were acquired using differential interference contrast (DIC), an FITC (fluorescein isothiocyanate) filter set, and a TRITC (tetramethylrhodamine isothiocyanate) filter set. Focal drift during the experiments was avoided using the autofocus system of the microscope.
For the analysis of the effect of tamoxifen on the phase partition and viability of the cells, cells were seeded in the presence of 4-hydroxytamoxifen in ethanol solvent (Sigma-Aldrich, St Louis, MO) or ethanol solvent control.
At each time of measurement, a transmission and two fluorescent images of the nuclei of the cells were acquired using differential interference contrast (DIC), an FITC (fluorescein isothiocyanate) filter set, and a TRITC (tetramethylrhodamine isothiocyanate) filter set. Focal drift during the experiments was avoided using the autofocus system of the microscope.
For the analysis of the effect of tamoxifen on the phase partition and viability of the cells, cells were seeded in the presence of 4-hydroxytamoxifen in ethanol solvent (Sigma-Aldrich, St Louis, MO) or ethanol solvent control.
3
Monitoring Cell Adaptation to Flow
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Cell adaptation to flow was monitored using an inverted Nikon‐Ti wide‐field microscope (Nikon, Japan) and an incubation chamber (Life Imaging Services, Switzerland). Both the flow bioreactor and the medium reservoir were maintained at a controlled temperature of 37 °C and CO2 concentration of 5%. Images were collected with a 20x, 0.45 NA long‐distance objective (Plan Fluor, Nikon, Japan). Time‐lapse experiments were set to routinely collect images, in different spatial positions of the sample, in the DAPI (nuclei) and TRITC (Golgi apparatus or fluorescent QDs) channel with a time resolution of 20 or 30 min.
VEC, Actin, and cell nuclei distribution were acquired in immunostained samples using a 60X, 1.4 NA oil immersion objective (Plan Fluor, Nikon, Japan), and the FITC, TRITC, and DAPI filter; respectively. Samples were imaged with an inverted Nikon‐Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU‐888 camera (Oxford Instruments, UK) and a pE‐100 LED illumination system (CoolLED Ltd, Andover, United Kingdom).
VEC, Actin, and cell nuclei distribution were acquired in immunostained samples using a 60X, 1.4 NA oil immersion objective (Plan Fluor, Nikon, Japan), and the FITC, TRITC, and DAPI filter; respectively. Samples were imaged with an inverted Nikon‐Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU‐888 camera (Oxford Instruments, UK) and a pE‐100 LED illumination system (CoolLED Ltd, Andover, United Kingdom).
4
Live Imaging of Cell Migration and Proliferation
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Following procedures described in (14 (link)), live imaging of the migration and proliferation processes was performed using an automated Nikon-Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU-888 camera (Oxford Instruments, UK) and a pE-100 light-emitting diode (LED) illumination system (CoolLED Ltd., Andover, UK) using a 20× or 10× objective (Plan Fluor, Nikon, Japan). An incubated chamber was used to maintain temperature and CO2 at 37°C and 5%, respectively (Life Imaging Services, Switzerland). Time-lapse recording was started approximately 1 hour after removing the MAts. The interval between image acquisition was 20 min to 1 hour, and a typical experiment lasted around between 24 to 48 hours. At the end of the experiments, the resulting time lapses for each set position were converted into individual 16-bit images for analysis. Fluorescent z-stacks of the signals emitted by the phospho-paxillin and paxillin at FAs were collected using a Nikon-Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU-888 camera (Oxford Instruments, UK) and selecting the optical filters based on the respective emission.
5
Cell Migration Monitoring with Microscopy
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Cell movement was monitored using an inverted Nikon‐Ti wide‐field microscope (Nikon, Japan) and an incubation chamber (Life Imaging Services, Switzerland). The medium was maintained at a controlled temperature of 37 °C and CO2 concentration of 5%. Images were collected with a 20×, 0.45 NA long‐distance objective (Plan Fluor, Nikon, Japan). Time‐lapse experiments were set to routinely collect images, in different spatial positions of the sample, in the BF channel with a time resolution of 20 min.
ZO‐1, tricellulin, EDU‐stained nuclei, and Hoechst or DAPI stained nuclei distribution were acquired in immunostained samples using a 60×, 1.4 NA oil immersion objective (Plan Fluor, Nikon, Japan), and the FITC, Cy5, TRITC, and DAPI filter respectively. Samples were images with an inverted Nikon‐Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU‐888 camera (Oxford Instruments, UK) and a pE‐100 LED illumination system (CoolLED Ltd., Andover, UK).
ZO‐1, tricellulin, EDU‐stained nuclei, and Hoechst or DAPI stained nuclei distribution were acquired in immunostained samples using a 60×, 1.4 NA oil immersion objective (Plan Fluor, Nikon, Japan), and the FITC, Cy5, TRITC, and DAPI filter respectively. Samples were images with an inverted Nikon‐Ti spinning disk confocal microscope (Nikon, Japan) equipped with an Andor DU‐888 camera (Oxford Instruments, UK) and a pE‐100 LED illumination system (CoolLED Ltd., Andover, UK).
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