A sensitive and versatile laser scanning confocal optical microscope for single-molecule fluorescence at 77 K

Hirschfeld, V.; Hübner, C. G.
November 2010
Review of Scientific Instruments;Nov2010, Vol. 81 Issue 11, p113705
Academic Journal
We developed a cryostat suitable for a laser scanning confocal microscope which allows for a short working distance and thus the usage of an objective with a high numerical aperture ensuring high collection efficiency. The in situ preparation of a thin layer of amorphous water is realized in a part of the cryostat, a Dewar vessel, which is put onto a custom-made, liquid-nitrogen immersed spin-coater. First tests on the setup are performed on a perylenemonoimide/polymethyl methacrylate model system using a standard oil objective and a dry objective at ambient temperature as well as a dry objective at liquid nitrogen temperature (77 K). Fluorescence resonance energy transfer (FRET) measurements on doubly labeled, freeze-quenched polyproline chains show the applicability of the new method on biomolecules. The alternating laser excitation (ALEX) is modified to a line-scanning process (slow ALEX) to optimize the sorting of the labeled molecules. Photophysics and photochemistry at liquid nitrogen temperature are investigated.


Related Articles

  • Mechanistic principles and applications of resonance energy transfer. Andrews, David L. // Canadian Journal of Chemistry;Sep2008, Vol. 86 Issue 9, p855 

    Resonance energy transfer is the primary mechanism for the migration of electronic excitation in the condensed phase. Well-known in the particular context of molecular photochemistry, it is a phenomenon whose much wider prevalence in both natural and synthetic materials has only slowly been...

  • Measurement of the V–T energy transfer rates of highly excited 2A1 NO2. Mazely, Troy L.; Friedl, Randall R.; Sander, Stanley P. // Journal of Chemical Physics;6/1/1994, Vol. 100 Issue 11, p8040 

    Production of electronic ground state NO2 (2A1) from 248 nm photolysis of HNO3 was detected by laser induced fluorescence (LIF). A growth in the LIF signal was observed following the photolysis and has been interpreted as the relaxation of NO2 through the higher vibrational levels of the X(2A1)...

  • [sup 3]He immersion cell for ultralow temperature study of amorphous solids. Rogge, Sven; Natelson, Douglas; Osheroff, D.D. // Review of Scientific Instruments;Apr97, Vol. 68 Issue 4, p1831 

    Describes the fabrication of a [sup 3]helium immersion cell for dielectric measurements of insulating amorphous solids in a nuclear demagnetization cryostat. Temperature range at the cryostat; Direct immersion of the samples in [sup 3]helium with two exchangers per electrode; Thermometer...

  • Photolysis of meta-azidophenol in solutions. Karyakina, L.; Oleinik, A. // High Energy Chemistry;Sep2007, Vol. 41 Issue 5, p350 

    The photolysis of meta-azidophenol in various organic solvents and aqueous solutions is studied by IR spectroscopy, electronic absorption spectroscopy, thin-layer chromatography (TLC), and analytical spot-test reactions. The polymeric derivatives of hydroxylamine are formed in organic solvents...

  • Time-domain sampling of x-ray pulses using an ultrafast sample response. Gaal, P.; Schick, D.; Herzog, M.; Bojahr, A.; Shayduk, R.; Goldshteyn, J.; Navirian, H. A.; Leitenberger, W.; Vrejoiu, I.; Khakhulin, D.; Wulff, M.; Bargheer, M. // Applied Physics Letters;12/10/2012, Vol. 101 Issue 24, p243106 

    We employ the ultrafast response of a 15.4 nm thin SrRuO3 layer grown epitaxially on a SrTiO3 substrate to perform time-domain sampling of an x-ray pulse emitted from a synchrotron storage ring. Excitation of the sample with an ultrashort laser pulse triggers coherent expansion and compression...

  • Cylindrical spike model for the formation of diamondlike thin films by ion deposition. Hofsäss, H.; Feldermann, H.; Merk, R.; Sebastian, M.; Ronning, C. // Applied Physics A: Materials Science & Processing;1998, Vol. 66 Issue 2, p153 

    Abstract. We present a new model for the formation of diamondlike films by ion deposition. In particular we model the observed ion energy dependence for the formation of tetrahedral amorphous carbon (ta-C). Ion deposition is treated as a cylindrical thermal spike, with energy loss along the ion...

  • Electronic coupling matrix element for electronic excitation energy transfer.... Harcourt, Richard D.; Ghiggino, Kenneth P. // Journal of Chemical Physics;7/22/1998, Vol. 109 Issue 4, p1310 

    Presents an expression for electronic coupling matrix element in electronic excitation energy transfer between equivalent chromophores. Observation on the vibronic coupling between S[sub 1] and S[sub 2] states; Emphasis of calculations on valence-shell ppi atomic orbitals; Consideration of a...

  • Direct study of energy transfer of vibrationally highly excited CS2 molecules. Dove, J. E.; Hippler, H.; Troe, J. // Journal of Chemical Physics;2/15/1985, Vol. 82 Issue 4, p1907 

    Collisional energy transfer at 300 K of vibrationally highly excited CS2 molecules (excitation energies 28 660 and 32 640 cm-1) is studied directly by time resolved UV absorption spectroscopy after laser excitation at 351 and 308 nm. Average energies 〈ΔE> transferred per collision for...

  • Rate expressions for excitation transfer. IV. Energy migration and superexchange phenomena. Scholes, Gregory D.; Ghiggino, Kenneth P. // Journal of Chemical Physics;11/22/1995, Vol. 103 Issue 20, p8873 

    General microscopic mechanisms of electronic excitation (energy) transfer (EET) in multichromophoric assemblies are investigated. Aspects of superexchange-mediated EET and energy migration (EM) and their contribution to the efficiency of donor-to-trap energy transport processes in macromolecules...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics