Posted by
Zdenek Svindrych-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Photostimulation-Andor-MicroPoint-or-Mosaic-tp7591009p7591011.html
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Hi Christian,
we have both on the same spinning disk (not a Dragonfly, though), but I
haven't used them for DNA damage.
The micropoint (basically a dye laser pumped by a nitrogen pulsed laser)
produces quite strong (50-100 uJ) nanosecond pulses that will ablate
anything. But in our configuration it can only do single shot ablation, the
laser is focused to a small spot on the sample, no galvo. The repetition
rate of the laser itself is limited to 15 Hz, so it may not be suitable for
your application.
Mosaic is a 100-200 mW 405 nm laser with a DMD projection, so arbitrary
patterns are possible, but if you want to irradiate a small point or thin
line, the power is of course much lower.
Mosaic can be controlled from MetaMorph (I haven't tried) or IQ. IQ should
be able to control Dragonfly quite well, I don't remember if there's any
critical functionality missing (maybe the ASI stage? Perfect Focus?). I use
the Fusion software almost exclusively. But IQ is also capable of grabbing
the screen output of another program (instead of getting data directly from
the camera), so it should be possible to run both IQ and Fusion
simultaneously, control the Mosaic through IQ and everything else through
Fusion...
Andor also offers (or used to offert) the FRAPPA - a galvo-based unit for
illuminating arbitrary shapes, this can deliver much stronger illumination
into small regions (point, line), but you need second (single mode) fiber
output from your laser module, and it's again controlled through IQ. I used
it with the good old Yokogawa X1, and since the FRAPPA sits in between the
spinning disk unit and the microscope stand, it's quite possible that it
limits your field of view...
Best, zdenek
On Wed, Jun 17, 2020 at 9:08 AM Christian Kukat <
[hidden email]>
wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> Post images on
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>
>
--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth