Laser cutters
The development of laser cutters started in the late 1990s. In a collaboration with A. A. “Tony” Hyman (now at MPI CBBG, Dresden) we initially intended to use optical tweezers to manipulate the movement of the posterior pole body in C. elegans. These experiments turned out to be quite inefficient and hence we decided to try a laser cutter built by PALM (now a part of Carl Zeiss, Jena) instead, which was on loan at EMBL for a few months.
Although we realized quite early on that its performance was not anywhere close to the diffraction limit we used it to complete the study (Grill et al., 2001). We then immediately devised a totally different device, which produced a diffraction limited spot of light, used a frequency tripled Nd:YAG instead of a nitrogen gas laser and exposed the specimen to about 0.1% of the energy of the commercial design (Grill et al., 2003).
When this device went to the MPI in Dresden, Julien Colombelli (now in Barcelona) built a new instrument, which was still in use at EMBL until early 2009. The laser cutters have been mainly used to perform relaxation experiments. They have been applied in various studies of microtubule behaviour in C. elegans (Grill above), in in vivo studies of microtubule shrinkage/growth (Colombelli et al., 2004; Colombelli et al., 2005), in studies of the role of zyxin by cutting actin fibres (Colombelli et al. 2009) and in studies related to the formation of the primordium in zebra fish (Gilmour et al., submitted). So far, no other instrument has created more collaborations than our laser cutters and again numerous careers are based on its development and application. One laser cutter has also been incorporated into a SPIM (Engelbrecht et al., 2007).
Publications
Colombelli J, Besser A, Kress H, Reynaud EG, Girard P, Caussinus E, Haselmann U, Small JV, Schwarz US, Stelzer EHK
Mechanosensing in actin stress fibers revealed by a close correlation between force and protein localization
Journal of Cell Science, 122:1665-1679 (2009).
Roukos V, Kotsantis P, Colombelli J, Kinkhabwala A, Taraviras S, Nishitani H, Stelzer EHK, Bastiaens P, Lygerou Z
The replication licensing factor CDT1 is dynamically recruited onto chromatin at sites of DNA damage
Febs Journal, 275(Suppl. 1):67 (2008).
Gilmour D, Haas P, Cakan G, Lecaucley V, Colombelli J, Stelzer EHK
Polarising migrating tissues during organogenesis
Developmental Biology, 319(2):488, (2008).
Colombelli J, Tängemo C, Haselmann U, Antony C, Stelzer EHK, Pepperkok R, Reynaud EG
A correlative light and electron microscopy method based on laser micropatterning and etching
Methods in Molecular Biology, Vol. 457, Membrane Trafficking edited by A. Vancura, Humana Press, p.203-214 (2008).
Colombelli J, Reynaud EG, Stelzer EHK
Investigating relaxation processes in cells and developing organisms: From cell ablation to cytoskeleton nanosurgery
Methods in Cell Biology (Edt. Berns M & Greulich KO), 82:267-291 (2007).
Gilmour D, Haas P, Cakan G, Lecaudey V, Valentin G, Colombelli J, Stelzer EHK
Pushing and pulling migrating tissues in vivo with GPCRs
FASEB J, 21:11 (2007).
Engelbrecht CJ, Greger K, Reynaud EG, Krzic U, Colombelli J, Stelzer EHK
Three-dimensional laser microsurgery in light-sheet based microscopy (SPIM)
Optics Express, 15(10) 6420 (2007).
Fink G, Schuchardt I, Colombelli J, Stelzer EHK, Steinberg G
Dynein-mediated pulling forces drive rapid mitotic spindle elongation in Ustilago maydis.
EMBO Journal 25(20),4897-908 (2006).
Colombelli J, Reynaud EG, Pepperkok R, Stelzer EHK
Laser nanosurgery in Cell Biology.
M-S Médecine Sciences 22(6-7), 651-658 (2006).
Colombelli J, Reynaud EG, Rietdorf J, Pepperkok R, Stelzer EHK
In vivo selective cytoskeleton dynamics quantification induced by pulsed UV laser nanosurgery.
Traffic 6(12),1093-102 (2005).
Colombelli J, Reynaud EG, Stelzer EHK
Subcellular nanosurgery with a pulsed subnanosecond UV-A laser.
Medical Laser Application 20: 217-222 (2005).
Colombelli J, Grill SW, Stelzer EHK
UV diffraction limited nanosurgery of live biological tissues.
Rev. Sci. Instr. 75(2): 472-478 (2004).
Colombelli J, Stelzer EHK
Diffraktionsbegrenzte Lasernanochirurgie.
GIT, 48(6): 600-602 (2004).
Grill SW, Howard J, Schäffer E, Stelzer EHK, Hyman AA
Diffraktionsbegrenzte Lasernanochirurgie.
Science, 301:518-521 (2003).
Grill SW, Gönczy P, Stelzer EHK, Hyman AA
Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo.
Nature, 409:630-633 (2001).
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