Targeted Ablation Using Laser Nanosurgery.
Laser-mediated dissection methods have been used for many years to micro-irradiate biological samples, but recent technological progress has rendered this technique more precise, powerful, and easy to use. Today pulsed lasers can be operated with diffraction limited, sub-micrometer precision to ablate intracellular structures. Here, we discuss laser nanosurgery setups and the instrumentation in our laboratory. We describe how to use this technique to ablate cytoskeletal elements in living cells. We also show how this technique can be used in multicellular organisms, to micropuncture and/or ablate cells of interest and finally how to monitor a successful laser nanosurgery.
Cell Biology and Cell Biophysics Unit, EMBL Heidelberg, Meyerhofstrasse 1, 69117, Heidelberg, Germany.
Naga Venkata
NV
Gayathri Vegesna
Cell Biology and Cell Biophysics Unit, EMBL Heidelberg, Meyerhofstrasse 1, 69117, Heidelberg, Germany.|Electron Microscopy Core Facility, EMBL Heidelberg, Meyerhofstrasse 1, 69117, Heidelberg, Germany.
Paolo
P
Ronchi
Cell Biology and Cell Biophysics Unit, EMBL Heidelberg, Meyerhofstrasse 1, 69117, Heidelberg, Germany.
Sevi
S
Durdu
Advanced Light Microscopy Facility, EMBL Heidelberg, Meyerhofstrasse 1, 69117, Heidelberg, Germany.
Stefan
S
Terjung
Cell Biology and Cell Biophysics Unit, EMBL Heidelberg, Meyerhofstrasse 1, 69117, Heidelberg, Germany. pepperko@embl.de.|Advanced Light Microscopy Facility, EMBL Heidelberg, Meyerhofstrasse 1, 69117, Heidelberg, Germany. pepperko@embl.de.
Rainer
R
Pepperkok
D000199:Actins / Q000378:metabolism; D000818:Animals; D015415:Biomarkers; D002138:Calibration; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D053685:Laser Therapy / Q000295:instrumentation / Q000379:methods*; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D008866:Microsurgery / Q000295:instrumentation / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods*; D015027:Zebrafish
D000199:Actins; D015415:Biomarkers
Laser ablation; Laser nanosurgery; Micropuncture; Photobleach; Subcellular structures