Merk, B. and Voss, K.-O. and Müller, I. and Fischer, B. E. and Jakob, B. and Taucher-Scholz, G. and Trautmann, C. and Durante, M. (2013):
Photobleaching setup for the biological end-station of the darmstadt heavy-ion microprobe.
In: Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms, Elsevier Science Publishing, pp. 81-84, 306, ISSN 0168583X,
[Online-Edition: http://dx.doi.org/10.1016/j.nimb.2012.11.043],
[Article]
Abstract
We report the upgrade of the epifluorescence microscope of the GSI heavy-ion microprobe with a galvo-scanned, 488 nm laser diode. The laser is focussed into the object plane by the water-immersion objective resulting in a focal spot size of about 1 μm. To increase temporal and spatial resolution a water-immersion objective with a high numerical aperture is integrated into the custom-build microscope. The upgraded system can now be used to bleach GFP-tagged proteins recruited to DNA damage induced by targeted single-ion irradiation. The system is demonstrated on NIH 3T3 cells with Ku80-GFP ion-targeted in heterochromatic and euchromatic DNA. Fluorescence recovery after photobleaching (FRAP) is shown to be significantly slower in heterochromatin.
| Item Type: | Article |
|---|---|
| Erschienen: | 2013 |
| Creators: | Merk, B. and Voss, K.-O. and Müller, I. and Fischer, B. E. and Jakob, B. and Taucher-Scholz, G. and Trautmann, C. and Durante, M. |
| Title: | Photobleaching setup for the biological end-station of the darmstadt heavy-ion microprobe |
| Language: | English |
| Abstract: | We report the upgrade of the epifluorescence microscope of the GSI heavy-ion microprobe with a galvo-scanned, 488 nm laser diode. The laser is focussed into the object plane by the water-immersion objective resulting in a focal spot size of about 1 μm. To increase temporal and spatial resolution a water-immersion objective with a high numerical aperture is integrated into the custom-build microscope. The upgraded system can now be used to bleach GFP-tagged proteins recruited to DNA damage induced by targeted single-ion irradiation. The system is demonstrated on NIH 3T3 cells with Ku80-GFP ion-targeted in heterochromatic and euchromatic DNA. Fluorescence recovery after photobleaching (FRAP) is shown to be significantly slower in heterochromatin. |
| Journal or Publication Title: | Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms |
| Volume: | 306 |
| Publisher: | Elsevier Science Publishing |
| Uncontrolled Keywords: | Microprobe, Live cell imaging, FRAP, Ku80 |
| Divisions: | 11 Department of Materials and Earth Sciences > Material Science > Ion-Beam-Modified Materials 11 Department of Materials and Earth Sciences > Material Science 11 Department of Materials and Earth Sciences |
| Date Deposited: | 21 Mar 2014 13:13 |
| Official URL: | http://dx.doi.org/10.1016/j.nimb.2012.11.043 |
| Identification Number: | doi:10.1016/j.nimb.2012.11.043 |
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