Merk, B. ; Voss, K.-O. ; Müller, I. ; Fischer, B. E. ; Jakob, B. ; Taucher-Scholz, G. ; Trautmann, C. ; 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, 306
doi: 10.1016/j.nimb.2012.11.043
Artikel, Bibliographie
Kurzbeschreibung (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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2013 |
| Autor(en): | Merk, B. ; Voss, K.-O. ; Müller, I. ; Fischer, B. E. ; Jakob, B. ; Taucher-Scholz, G. ; Trautmann, C. ; Durante, M. |
| Art des Eintrags: | Bibliographie |
| Titel: | Photobleaching setup for the biological end-station of the darmstadt heavy-ion microprobe |
| Sprache: | Englisch |
| Publikationsjahr: | 1 Juli 2013 |
| Verlag: | Elsevier Science Publishing |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms |
| Jahrgang/Volume einer Zeitschrift: | 306 |
| DOI: | 10.1016/j.nimb.2012.11.043 |
| Kurzbeschreibung (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. |
| Freie Schlagworte: | Microprobe, Live cell imaging, FRAP, Ku80 |
| Fachbereich(e)/-gebiet(e): | 11 Fachbereich Material- und Geowissenschaften > Materialwissenschaft > Fachgebiet Ionenstrahlmodifizierte Materialien 11 Fachbereich Material- und Geowissenschaften > Materialwissenschaft 11 Fachbereich Material- und Geowissenschaften |
| Hinterlegungsdatum: | 21 Mär 2014 13:13 |
| Letzte Änderung: | 21 Mär 2014 13:13 |
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