Laser scanning confocal microscopy system
Final Report Abstract
In the last three years, the confocal microscope became an integral and indispensible part of a variety of projects within and outside of our department. Its usage includes a variety of live cells applications and conventional fixed specimen imaging. Internal and external users belonging to multiple departments and disciplines (Physics, Chemistry, Biology) use the system for their research projects. One of the key research in which the instrument has contributed very significantly is our interdisciplinary DFG funded research college 1657 „Molecular and cellular responses to ionizing radiation”. This is part of our department’s research focus on biology of stress responses (http://www.bio.tudarmstadt.de/ag/forschen/schwerpunkte.en.jsp) and includes groups working on microorganisms, to plants and humans. Animal models as well as cellular models are employed here to elucidate the basis of chromosome stability in normal proliferating cells as well as tumor cells and in response to different types of stress, including radiation stress use in therapy. One o the focus here has been to study the kinetics of DNA damage and its repair using live cell imaging combined with laser microirradiation. Other research areas cover the response of cells and organs to exogenous signals and the signal reception at the level of the cell membrane. As a counterpoint also synthetic membranes assembled into artificial vesicles have been analyzed. To improve drug delivery, collaborative projects in the interface between chemistry and biology have used live-cell microscopy to quantify the kinetics of uptake of drugs as well as their function in vivo. In several of the above mentioned research projects, dynamic live cell measurements have been performed and include: fluorescence photobleaching and photoactivation techniques, laser microbeam irradiation and ratiometric measurements (e.g., of radical oxygen species). These experiments have encompassed from synthetic artificial vesicles, stretched DNA molecules, to bacterial microfilms, insects, plants and mammalian cells and organisms. This is made possible by to the large variety of objectives at the microscope, from very low to the highest magnification. In addition, very complex live-cell imaging experiments (already described in the application) have been successfully performed, which is feasible due to the multiple software modules purchased with the system. Our present directions expand into multi color superresolution light microscopy in fixed and living cells to bridge the gap to electron microscopy.
Publications
- (2011). Histone acetylation controls the inactive X chromosome replication dynamics. Nat. Commun. 2: 222
Casas-Delucchi, C. S., Brero, A., Rahn, H.-P., Solovei, I., Wutz, A., Cremer, T., Leonhardt, H. and Cardoso, M. C.
(See online at https://doi.org/10.1038/ncomms1218) - (2012). Biofilm formation by haloarchaea. Environ Microbiol 14: 3159-3174
Fröls, S., Dyall-Smith, M. and Pfeifer, F.
- (2012). ERES (ER exit sites) and the “Secretory Unit Concept.” J. Microsc. 247: 48–59
Langhans, M., Meckel, T., Kress, A., Lerich, A. and Robinson, D. G.
- (2012). Histone hypoacetylation is required to maintain late replication timing of constitutive heterochromatin. Nucleic Acids Res. 40: 159- 169
Casas-Delucchi, C. S., van Bemmel, J. G., Haase, S., Herce, H. D., Nowak, D., Meilinger, D., Stear, J. H., Leonhardt, H. and Cardoso, M. C.
(See online at https://doi.org/10.1093/nar/gkr723) - (2012). Intricate paths of cells and networks becoming "Cholinergic" in the embryonic chicken retina. J. Comp. Neurol. 520:3181-93
Thangaraj, G., Greif, A., Bachmann, G. and Layer, P. G.
- (2012). Structural organization of DNA in Chlorella viruses. PLoS One 7:e30133
Wulfmeyer, T. Polzer, C. Hiepler, G., Hamacher, K., Shoeman, R., Dunigan, D. Van Etten, J. L., Lolicato, M., Moroni, A., Thiel, G. and Meckel, T.
- (2012). The Relevance of Lysine Snorkeling in the Outer Transmembrane Domain of Small Viral Potassium Ion Channels. Biochemistry 51, 5571-5579
Gebhardt, M., Henkes, L., Tayefeh, S., Hertel, B., Greiner, T., Van Etten, J., Baumeister, D., Cosentino, C., Moroni, A., Kast, S. and Thiel, G.
- (2013). Archaeal biofilms: widespread and complex. Biochem Soc Trans 41: 393-398
Fröls, S.
- (2013). Viral encoded potassium ion channel is a structural protein in the Chlorovirus Paramecium bursaria chlorella virus-1 (PBCV-1) virion. J. Gen. Virol. 94:2549-56
Romani, G., Piotrowski, A., Hilmer, S., Gurnin, J, VanEtten, J. L., Moroni, A., Thiel, G. and Hertel, B.
- (2014). Analysis of fluid-phase endocytosis in (intact) plant cells. Methods Mol. Biol. 1209: 45–61
Bandmann, V., Haub, P. and Meckel, T.
(See online at https://doi.org/10.1007/978-1-4939-1420-3_4) - (2014). Application and possible mechanisms of combining LLLT (low level laser therapy), infrared hyperthermia and ionizing radiation in the treatment of cancer. SPIE 8932, Mechanisms for Low-Light Therapy IX, 893202
Abraham, E. H., Woo, Van H., Harlin-Jones, C., Heselich, A. and Frohns, F.
(See online at https://doi.org/10.1117/12.2038630) - (2014). Gelatine methacrylamide-based hydrogels: an alternative threedimensional cancer cell culture system. Acta Biomater. 10: 2551–62
Kaemmerer, E., Melchels, F. P. W., Holzapfel, B. M., Meckel, T., Hutmacher, D. W. and Loessner, D.
(See online at https://doi.org/10.1016/j.actbio.2014.02.035) - (2014). Inefficient doublestrand break repair in murine rod photoreceptors with inverted heterochromatin organization. Curr Biol. 24: 1080-1090
Frohns, A., Frohns, F., Naumann, S. C., Layer, P. G. and Löbrich, M.
(See online at https://doi.org/10.1016/j.cub.2014.03.061) - (2014). Single-molecule detection and tracking in plants. Protoplasma 251:277-91
Langhans, M. and Meckel, T.
(See online at https://doi.org/10.1007/s00709-013-0601-0) - (2014). The sorting of a small potassium channel in mammalian cells can be shifted between mitochondria and plasma membrane. Cell Calcium
v. Charpuis, C., Meckel, T., Moroni, A. and Thiel, G.
(See online at https://doi.org/10.1016/j.ceca.2014.09.009) - Single molecule microscopy in 3D cell cultures and tissues. Adv. Drug Deliv. Rev. Volumes 79–80, 15 December 2014, Pages 79-94
Lauer, F., Kaemmerer, E. and Meckel, T.
(See online at https://doi.org/10.1016/j.addr.2014.10.008) - Low-dose photon irradiation alters cell differentiation via activation of hIK channels. Pflügers Archiv - European Journal of Physiology, August 2015, Volume 467, Issue 8, pp 1835–1849
Roth, B., Gibhardt , C. S. Becker, P., Gebhardt, M. Knoop, J., Fournier, C., Moroni, A. and Thiel, G.
(See online at https://dx.doi.org/10.1007/s00424-014-1601-4)
