Posted by
PAVAK SHAH on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Photostimulation-Andor-MicroPoint-or-Mosaic-tp7591009p7591010.html
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Hi Christian,
The Mosaic is really the system intended for patterns, although I don't
have much experience with it. It's a projector, so you may trade off a
little bit of sharpness in your pattern and some limitation in peak
intensity for truly simultaneous illumination.
The micropoint is a dye laser that maxes out at a 15 Hz repetition rate,
not really practical for quickly painting shapes.
A few vendors sell 355 nm pulse lasers with kHz rep rates that can paint
shapes well, but I don't know how software integration works with a
Dragonfly. Gataca's iLas and Rapp's Firefly systems come to mind. I moved
from a micropoint to a Rapp system and have been extremely impressed with
its capabilities. 355 nm illumination will also produce DNA damage more
readily than 405 nm. We mostly do point ablations, but it also handles
painting very well.
Best,
Pavak
On Wed, Jun 17, 2020, 6:09 AM Christian Kukat <
[hidden email]>
wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
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http://www.imgur.com and include the link in your posting.
> *****
>
> 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>
>