Cells were plated in glass bottom 35 mm-petri dishes for 24 h (No. 1.5, MatTeK Corporation, United States). Intracellular Ca2+ labeling and imaging was described as before (Chen et al., 2019 (link)). In details, cells were labeled with 5 μm Fura-2 AM (F1221, Thermo Fisher Scientific, United States) supplied with 0.02% Pluronic F-127 (P3000MP, Thermo Fisher Scientific, United States) and then were illuminated at an alternating excitation wavelengths of 340 nm and 380 nm in an Epi-fluorescence microscope with a Plan-Fluor 40×/1.3 Oil objective (Eclipse Ti, Nikon, Japan). The emitted fluorescence was recorded at 510 nm with an Andor Zyla sCMOS camera (Oxford Instruments, United Kingdom). Exposure time was typically 100–200 ms, and images were collected every 10–20 s. Images were analyzed using MetaFluor software (Universal Imaging Corporation, United States). Fluorescent images were background corrected and cells with similar fluorescence intensity were analyzed.
Andor zyla scmos camera
Manufactured by Oxford Instruments
Sourced in United Kingdom
The Andor Zyla sCMOS camera is a high-performance scientific camera designed for a variety of imaging applications. It features a large active area, high resolution, and fast readout speeds. The camera utilizes sCMOS (scientific Complementary Metal-Oxide-Semiconductor) technology to provide low noise, high dynamic range, and high quantum efficiency.
Automatically generated - may contain errors
Lab products found in correlation
Fura 2 am, by Thermo Fisher Scientific (1 mentions)
Pluronic f 127, by Thermo Fisher Scientific (1 mentions)
Plan fluor 40 1.3 oil objective eclipse ti, by Nikon (1 mentions)
Metafluor software, by Universal Imaging (1 mentions)
Te2000e pfs microscope, by Nikon (1 mentions)
Nis elements software, by Nikon (1 mentions)
Ti e inverted microscope, by Nikon (1 mentions)
3 protocols using andor zyla scmos camera
1
Intracellular Calcium Imaging with Fura-2
2
Live-cell imaging of cell mitosis
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Cells were seeded onto 24- or 96-well cell culture plates, and the plates were placed onto a TE2000E-PFS microscope (Nikon, Tokyo, Japan) equipped with an Andor Zyla sCMOS camera (Oxford Instruments, UK) and a Chamlide TC temperature, humidity, and CO2 control chamber (Live Cell Instrument, Seoul, South Korea). Images were captured every 5 min for 24 h or every 10 min for 48 h. Data acquisition was carried out with Metamorph 7.8.6 software (Molecular Devices, Sunnyvale, CA, USA) and analysis was performed using ImageJ software (National Institutes of Health, Bethesda, MD, USA). For histone H2B-GFP-expressing cells, the duration of mitosis was estimated from the time of DNA condensation to decondensation. For FUCCI-expressing cells, the duration of mitosis was estimated from the time of the APC/C reporter dispersing from the nucleus (due to nuclear envelope breakdown) to its destruction.
3
Quantifying Pigmentation in Eye Tissues
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We did not obtain clear anterior and posterior landmarks together in the same histologic sections, so we used different sections to quantify anterior and posterior pigmentation for each eye. For pigmentation analyses of anterior structures, we selected 3 sections from each eye that included the approximate center of the lens. Images were taken of the iris pigmented epithelium, the ciliary body, and the ora serrata (for anterior RPE analysis). For pigmentation analyses of the posterior RPE, we selected 2-3 sections containing the optic nerve, and images were taken of the RPE on both sides adjacent to the optic nerve. Sections were imaged using a Nikon Ti-E inverted microscope (Nikon Instruments Inc, Melville, NY) with a high-sensitivity Andor Zyla sCMOS camera (Oxford Instruments, Abingdon, Oxon, United Kingdom) and LED light source. For the IPE, anterior RPE, and posterior RPE images were taken using 100x magnification, and Z-stacks of each section were taken using 0.02-µm steps through the tissue layers containing pigmented melanosomes using Nikon NIS-Elements software. For the ciliary body, images were taken on the same microscope using 20x magnification and 0.2 µm steps. Stacks were merged to create an Extended Depth of Focus (EDF) file. EDF files were then exported to create a TIFF file for image analysis.
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