Cells were grown on chambered cover glasses to 70–80% confluency, at which time cells were stimulated with IL-1β (1 μg/ml) or TNFα (1 μg/ml). Live cell images were immediately captured every 10 seconds for 1 hour (IL-1β stimulation) or 30 min (TNFα stimulation) under an Andor spinning disk confocal microscope system equipped with a Nikon Ti motorized microscope (with 60× oil objective), a CSUX1 Spinning Disk Confocal head (Yokogawa), an Andor iXon EMCCD camera and a Neo sCMOS camera. Images were analyzed by ImageJ. To quantify the number of cellular condensates, backgrounds of images were first subtracted by using the module of subtract background on ImageJ, and then condensates (~ 0.3–1 μm in diameter) in cells were spotted and counted using the plugin of SpotCounter on ImageJ (the threshold of fluorescent intensity in detecting NEMO puncta was set above that of free NEMO). For tracking the fusion of two condensates as shown in Figure 2E –F , trajectories and positions of condensates were obtained using the plugin of TrackMate on ImageJ. Cells that contained NEMO puncta after IL-1β or TNFα treatment represented > 90% of stimulated cells.
Ti motorized microscope
Manufactured by Nikon
The Nikon Ti motorized microscope is a high-performance research-grade instrument designed for advanced imaging and analysis applications. It features a motorized stage and focus control, allowing for precise and automated sample positioning and focusing. The microscope is equipped with advanced optics and illumination systems, providing researchers with clear, high-resolution images for a variety of applications.
Automatically generated - may contain errors
2 protocols using ti motorized microscope
1
Imaging NEMO Condensate Dynamics
2
Imaging NEMO Condensate Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were grown on chambered cover glasses to 70–80% confluency, at which time cells were stimulated with IL-1β (1 μg/ml) or TNFα (1 μg/ml). Live cell images were immediately captured every 10 seconds for 1 hour (IL-1β stimulation) or 30 min (TNFα stimulation) under an Andor spinning disk confocal microscope system equipped with a Nikon Ti motorized microscope (with 60× oil objective), a CSUX1 Spinning Disk Confocal head (Yokogawa), an Andor iXon EMCCD camera and a Neo sCMOS camera. Images were analyzed by ImageJ. To quantify the number of cellular condensates, backgrounds of images were first subtracted by using the module of subtract background on ImageJ, and then condensates (~ 0.3–1 μm in diameter) in cells were spotted and counted using the plugin of SpotCounter on ImageJ (the threshold of fluorescent intensity in detecting NEMO puncta was set above that of free NEMO). For tracking the fusion of two condensates as shown in Figure 2E –F , trajectories and positions of condensates were obtained using the plugin of TrackMate on ImageJ. Cells that contained NEMO puncta after IL-1β or TNFα treatment represented > 90% of stimulated cells.
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!