Posted by
Christian Kukat on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Photostimulation-Andor-MicroPoint-or-Mosaic-tp7591009.html
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Dear confocal list community,
We have an Andor Dragonfly spinning disk confocal, but no FRAP scanner, yet.
We are interested in performing experiments to recruit fuorescently-tagged proteins to DNA damage sites using laser microirradiation and live-cell imaging. For this, we plan to microirradiate points but also line patterns, as described for example in the publications mentioned below (below signature). We would prefer 405 nm for the wavelenght to induce DNA damage.
Andor offered to us now 2 different tools for photostimulation:
- MicroPoint
https://andor.oxinst.com/products/photostimulation/micropoint <
https://andor.oxinst.com/products/photostimulation/micropoint>
- Mosaic
https://andor.oxinst.com/products/photostimulation/mosaic <
https://andor.oxinst.com/products/photostimulation/mosaic>
We have now several questions and hope that someone of you can help us:
- Does anybody have experiences with one or the other tool regarding the recruitment of proteins to DNA damage sites?
- How fast is the MicroPoint system for the creation of line patterns?
- As far as we know, none of them is integrated in the Andor Fusion software. So how well works this using the iQ software or swapping between Fusion and iQ?
- What are the pros and cons of them?
We appreciate any input from the community!
Best wishes and stay safe!
Christian
_______________________________________________________________________
Christian Kukat, PhD
Head of FACS & Imaging Core Facility
Max Planck Institute for Biology of Ageing
- ISAC SRL Emerging Leader 2016-2020 -
Joseph-Stelzmann-Str. 9b, D-50931 Köln / Cologne, Germany
E-mail:
[hidden email]
www.age.mpg.de
_______________________________________________________________________
Examples in publications:
Fig. 1 in Kim J.J., Kumbhar R., Gong F., Miller K.M. (2019) In Time and Space: Laser Microirradiation and the DNA Damage Response. In: Balakrishnan L., Stewart J. (eds) DNA Repair. Methods in Molecular Biology, vol 1999. Humana, New York, NY
https://link.springer.com/protocol/10.1007/978-1-4939-9500-4_3#Sec14 <
https://link.springer.com/protocol/10.1007/978-1-4939-9500-4_3#Sec14>
Fig. 1 in Tampere, M. and Mortusewicz, O. (2016). DNA Damage Induction by Laser Microirradiation. Bio-protocol 6(23): e2039. DOI: 10.21769/BioProtoc.2039 <
https://doi.org/10.21769/BioProtoc.2039>.
https://bio-protocol.org/e2039 <
https://bio-protocol.org/e2039>
Fig. 2 in Haince JF, McDonald D, Rodrigue A, et al. PARP1-dependent kinetics of recruitment of MRE11 and NBS1 proteins to multiple DNA damage sites. J Biol Chem. 2008;283(2):1197-1208. doi:10.1074/jbc.M706734200
https://www.jbc.org/content/283/2/1197.full <
https://www.jbc.org/content/283/2/1197.full>