For time-lapse imaging, U2OS cells expressing EB3-mNeonGreen were grown on a 35 mm µ-Dish with a polymer coverslip bottom (Ibidi GmbH, Gräfelfing, Germany). Prior to imaging, the medium was replaced for FluoroBrite™ DMEM, supplemented with 25 mM HEPES and 1% FCS, 30 min before imaging. Time-lapse sequences were collected in seven optical slices (0.1 µm steps) for 1 min at 1 s interval with the Andor Dragonfly 503 spinning disc confocal system (Oxford Instruments, Abingdon, UK) equipped with a stage top microscopy incubator (Okolab, Ottaviano, Italy), 488 nm solid-state 150 mW laser, HCX PL APO 63×/1.4 oil objective, and Zyla 4.2 PLUS sCMOS camera. For each experiment, at least 10 cells were imaged (acquisition parameters: 40 µm pinhole size, 15% laser power, 50 ms exposure time, 525/50 nm emission filter). The time-lapse sequences were deconvoluted with Huygens Professional software v. 19.04 (Scientific Volume Imaging, Hilversum, The Netherlands), and maximum intensity projection of z stack was made for each time point in Fiji. Newly nucleated microtubules were detected by manual counting of EB3 comets emanating from the centrosomes.
Stage top microscopy incubator
Manufactured by Okolab
Sourced in United Kingdom
The Stage top microscopy incubator is a compact environmental control system designed to maintain optimal temperature, humidity, and gas conditions for live cell imaging during microscopy experiments. It provides a stable and controlled environment to support the viability of cells and tissues under observation.
Automatically generated - may contain errors
2 protocols using stage top microscopy incubator
1
Visualizing Microtubule Nucleation in U2OS Cells
2
Monitoring Microtubule Nucleation in U2OS Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For time-lapse imaging, U2OS cells expressing EB3-mNeonGreen were grown on a 35 mm µ-Dish with ibidi polymer coverslip bottom (Ibidi GmbH, Gräfelfing, Germany). Prior to imaging, the medium was replaced for FluoroBrite™ DMEM (ThermoFisher Scientific, Waltham, MA), supplemented with 25 mM HEPES and 1% FCS, 30 min before imaging. Time-lapse sequences were collected in seven optical slices (0.1 µm steps) for 1 min at 1 s interval with the Andor Dragonfly 503 spinning disc confocal system (Oxford Instruments, Abingdon, UK) equipped with a stage top microscopy incubator (Okolab, Ottaviano, Italy), 488 nm solid-state 150 mW laser, HCX PL APO 63x oil objective, NA 1.4, and Zyla sCMOS 16 bit camera. For each experiment, at least 10 cells were imaged (acquisition parameters: 40 µm pinhole size, 15% laser power, 50 ms exposure time, 525/50 nm emission filter). The time-lapse sequences were deconvoluted with Huygens Professional software v. 19.04 (Scientific Volume Imaging, Hilversum, the Netherlands), and maximum intensity projection of z stack was made for each time point in Fiji. Newly nucleated microtubules were detected by manual counting of EB3 comets emanating from the centrosomes.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!