Telomere mobility analysis was done as described13 (link) with minor adjustments. TRF2ts cells expressing eGFP-TRF1 and mCherry-BP1-2 (53BP1 aa 1220-1711) were seeded on Willco wells and left to attach overnight at 32°C. Prior to imaging, cells were incubated at 37°C for 30 min in Leibovitz’s L15 medium inside the Microscope climate chamber. Cells were monitored every minute for 20 min (t=20 frames) at 37°C. 5 μm Z-stacks at 0.5 μm steps in both eGFP and mCherry channels were obtained with a 63x 1.4 oil objective using ZEN software on an Observer Z1 microscope system (Carl Zeiss, Germany). Images were obtained at 2×2 binning with 512×512 pixels. Tracking of telomeres was performed with FIJI software, cells were registered with the Rigid body option from the StackReg plugin. Particles were then tracked with the particle detector and tracker plugin from Mosaic. Telomeres to track were selected on basis of colocalization with mCherry-BP1-2, and tracked for at least 17 out of 20 frames.
Observer z1 microscope system
Manufactured by Zeiss
Sourced in Germany
The Observer Z1 microscope system is a high-performance optical microscope designed for a wide range of applications. It features advanced optical components and a modular design that allows for customization to meet specific research or analysis needs. The core function of the Observer Z1 is to provide users with high-quality imaging and observation capabilities for various samples and materials.
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Lab products found in correlation
2 protocols using observer z1 microscope system
1
Tracking Telomere Dynamics in Cells
2
Tracking Telomere Dynamics in Cells
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Telomere mobility analysis was done as described13 (link) with minor adjustments. TRF2ts cells expressing eGFP-TRF1 and mCherry-BP1-2 (53BP1 aa 1220-1711) were seeded on Willco wells and left to attach overnight at 32°C. Prior to imaging, cells were incubated at 37°C for 30 min in Leibovitz’s L15 medium inside the Microscope climate chamber. Cells were monitored every minute for 20 min (t=20 frames) at 37°C. 5 μm Z-stacks at 0.5 μm steps in both eGFP and mCherry channels were obtained with a 63x 1.4 oil objective using ZEN software on an Observer Z1 microscope system (Carl Zeiss, Germany). Images were obtained at 2×2 binning with 512×512 pixels. Tracking of telomeres was performed with FIJI software, cells were registered with the Rigid body option from the StackReg plugin. Particles were then tracked with the particle detector and tracker plugin from Mosaic. Telomeres to track were selected on basis of colocalization with mCherry-BP1-2, and tracked for at least 17 out of 20 frames.
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