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. © 2013 Elsevier B.V. All rights reserved.
Photobleaching setup for the biological end-station of the darmstadt heavy-ion microprobe / Merk, B.; Voss, K. -O.; Muller, I.; Fischer, B. E.; Jakob, B.; Taucher-Scholz, G.; Trautmann, C.; Durante, M.. - In: NUCLEAR INSTRUMENTS & METHODS IN PHYSICS RESEARCH. SECTION B, BEAM INTERACTIONS WITH MATERIALS AND ATOMS. - ISSN 0168-583X. - 306:(2013), pp. 81-84. [10.1016/j.nimb.2012.11.043]
Photobleaching setup for the biological end-station of the darmstadt heavy-ion microprobe
Durante M.
2013
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. © 2013 Elsevier B.V. All rights reserved.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
