An insert for single-molecule magnetic-resonance spectroscopy in an external magnetic field

Meer, Harmen van der; Disselhorst, J.A.J.M.; Köhler, Jürgen; Brouwer, A. C. J.; Groenen, E. J. J.; Schmidt, Jan

doi:10.1063/1.1146164

Cited by 26 publications

(8 citation statements)

References 9 publications

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“…Typically, fluorescence-excitation spectra of an individual complex are obtained in rapid succession by scanning the laser repetitively through the spectral range of interest and storing the individual traces separately in computer memory. Details about the experimental setup can be found in Van der Meer et al (1995) and Hofmann et al (2004b).…”

Section: Methodsmentioning

confidence: 99%

Low-temperature single-molecule spectroscopy on photosynthetic pigment–protein complexes from purple bacteria

Oellerich

Köhler

2009

Photosynth Res

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The primary reactions of purple bacterial photosynthesis take place within two well characterized pigment-protein complexes, the core Reaction Center-Light Harvesting 1 (RC-LH1) complex and the more peripheral Light Harvesting 2 (LH2) complex. These antenna complexes serve to absorb incident solar radiation and to transfer it to the reaction-centers, where it is used to 'power' the photosynthetic redox reaction. This review provides an overview of how the character of the electronically excited states of these pigment-protein complexes are determined by quantum mechanics and how the respective spectral signatures can be observed by single-molecule spectroscopy.

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Section: Methodsmentioning

confidence: 99%

Low-temperature single-molecule spectroscopy on photosynthetic pigment–protein complexes from purple bacteria

Oellerich

Köhler

2009

Photosynth Res

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“…Next, the sample was mounted into a home‐built cryogenic insert which also accommodated the microscope objective (NA, numerical aperture = 0.90, Microthek, Hamburg, Germany). The insert was based on a mechanical arrangement, described previously by van der Meer et al . (1995), that allows in situ adjustments of the objective and sample with respect to each other along two mutually orthogonal directions (focus and sample position) with submicrometre precision.…”

Section: Methodsmentioning

confidence: 99%

Epifluorescence, confocal and total internal reflection microscopy for single‐molecule experiments: a quantitative comparison

Lang

Baier

Köhler

2006

Journal of Microscopy

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Summary Epifluorescence, confocal and total internal reflection microscopy are the most widely used techniques for optical single‐molecule experiments. Employing these methods, we recorded the emission intensity of the same single molecule as a function of the excitation rate under otherwise identical experimental conditions. Evaluation of these data provides a quantitative comparison of the signal‐to‐background ratios that can be achieved for the three microscopic techniques.

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“…When slightly tilted around either hinge, the lens-mirror holder also shifts in the sample plane. Our group has previously shown that flexure hinges provide reliable and stable positioning in low-temperature microscopy (27,28).…”

Section: Inside the Cryostatmentioning

confidence: 99%

Laser-Driven Microsecond Temperature Cycles Analyzed by Fluorescence Polarization Microscopy

Zondervan

Kulzer

Meer

et al. 2006

Biophysical Journal

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We demonstrate a novel technique to achieve fast thermal cycles of a small sample (a few femtoliters). Modulating a continuous near-infrared laser focused on a metal film, we can drive the local temperature from 130 to 300 K and back, within a few microseconds. By fluorescence microscopy of dyes in a thin glycerol film, we record images of the hot spot, calibrate its temperature, and follow its variations in real time. The temperature dependence of fluorescence anisotropy, due to photophysics and rotational diffusion, gives a steady-state temperature calibration between 200 and 350 K. From 200 to 220 K, we monitor temperature more accurately by fluorescence autocorrelation, a probe for rotational diffusion. Time-resolved measurements of fluorescence anisotropy give heating and cooling times of a few microseconds, short enough to supercool pure water. We designed our method to repeatedly cycle a single (bio)molecule between ambient and cryostat temperatures with microsecond time resolution. Successive measurements of a structurally relevant variable will decompose a dynamical process into structural snapshots. Such temperature-cycle experiments, which combine a high time resolution with long observation times, can thus be expected to yield new insights into complex processes such as protein folding.

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An insert for single-molecule magnetic-resonance spectroscopy in an external magnetic field

Abstract: Articles you may be interested inSimultaneous measurements of heat capacity and spinlattice relaxation time in high magnetic field at low temperature Rev.A sensitive capacitance thermometer at low temperature for use in magnetic fields up to 20 T Rev.

Cited by 26 publications

References 9 publications

Low-temperature single-molecule spectroscopy on photosynthetic pigment–protein complexes from purple bacteria

Low-temperature single-molecule spectroscopy on photosynthetic pigment–protein complexes from purple bacteria

Epifluorescence, confocal and total internal reflection microscopy for single‐molecule experiments: a quantitative comparison

Laser-Driven Microsecond Temperature Cycles Analyzed by Fluorescence Polarization Microscopy

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