2-GHz frequency-domain fluorometer

We developed a frequency‐domain fluorometer which operates from 4 to 2000 MHz. The modulated excitation is provided by the harmonic content of a laser pulse train (3.76 MHz, 5 ps) from a synchronously pumped and cavity dumped dye laser. The phase angle and modulation of the emission are measured with a microchannel‐plate photomultiplier (PMT). Cross‐correlation detection is performed outside the PMT. The high‐frequency signals for cross correlation were obtained by multiplication of the output from a 500‐MHz frequency synthesizer. The performance was verified in several ways, including measurement of known time delays and examination of standard fluorophores. The detector displayed no detectable color effect, with the 300–600‐nm difference being less than 5 ps. The precision of the measurements is adequate to detect differences of 20 ps for decay times of 500 ps. A correlation time of 53 ps was found for indole in water at 20 °C. The shortest correlation time we measured was 15 ps for indole in methanol/w...

[1]  B. Maliwal,et al.  Construction and performance of a variable-frequency phase-modulation fluorometer. , 1985, Biophysical chemistry.

[2]  J. Lakowicz,et al.  Correction of timing errors in photomultiplier tubes used in phase-modulation fluorometry. , 1981, Journal of biochemical and biophysical methods.

[3]  A. McKinnon,et al.  Correction of instrumental time response variation with wavelength in fluorescence lifetime determinations in the ultraviolet region , 1977 .

[4]  E. Gratton,et al.  A continuously variable frequency cross-correlation phase fluorometer with picosecond resolution. , 1983, Biophysical journal.

[5]  I. Yamazaki,et al.  Microchannel‐plate photomultiplier applicability to the time‐correlated photon‐counting method , 1985 .

[6]  W. Ware,et al.  Kinetics of diffusion‐controlled reactions: Transient effects in fluorescence quenching , 1975 .

[7]  J. Mccammon,et al.  Dynamics of Proteins and Nucleic Acids , 2018 .

[8]  S. Kinoshita,et al.  Picosecond Fluorescence Spectroscopy By Time-Correlated Single-Photon Counting , 1985 .

[9]  G. K. Rollefson,et al.  The Determination of the Fluorescence Lifetimes of Dissolved Substances by a Phase Shift Method , 1953 .

[10]  P. Bayley,et al.  Spectroscopy and the dynamics of molecular biological systems , 1985 .

[11]  E Gratton,et al.  Resolution of mixtures of fluorophores using variable-frequency phase and modulation data. , 1984, Biophysical journal.

[12]  James N. Demas,et al.  Excited State Lifetime Measurements , 1983 .

[13]  I. Munro,et al.  Time Resolved Fluorescence Spectroscopy With Synchrotron Radiation , 1985 .

[14]  R. Lumry,et al.  High Performance Phase Fluorometer Constructed from Commercial Subunits , 1965 .

[15]  D. O'connor,et al.  Time-Correlated Single Photon Counting , 1984 .

[16]  J. Lakowicz,et al.  Time-resolved fluorescence emission spectra of labeled phospholipid vesicles, as observed using multi-frequency phase-modulation fluorometry , 1984 .

[17]  W. Ware,et al.  The direct observation of transient effects in diffusion-controlled fluorescence quenching , 1973 .

[18]  P. R. Bevington,et al.  Data Reduction and Error Analysis for the Physical Sciences , 1969 .

[19]  R. R. Alfano,et al.  Biological events probed by ultrafast laser spectroscopy , 1982 .

[20]  J. Lakowicz,et al.  Measurement of subnanosecond anisotropy decays of protein fluorescence using frequency-domain fluorometry. , 1986, The Journal of biological chemistry.

[21]  J. Lakowicz Principles of fluorescence spectroscopy , 1983 .

[22]  B. Maliwal,et al.  Time-resolved fluorescence anisotropies of diphenylhexatriene and perylene in solvents and lipid bilayers obtained from multifrequency phase-modulation fluorometry. , 1985, Biochemistry.

[23]  J. R. Wilson,et al.  Optoelectronics, an introduction , 1984 .

[24]  J. Lakowicz,et al.  Analysis of fluorescence decay kinetics from variable-frequency phase shift and modulation data. , 1984, Biophysical journal.

[25]  A. Persoons,et al.  Fluorescence lifetime resolution with phase fluorimetry , 1983 .

[26]  E. Gratton,et al.  Measuring fluorescence decay times by phase-shift and modulation techniques using the high harmonic content of pulsed light sources , 1980 .

[27]  G. Heidt,et al.  Phase fluorometer with a continuously variable frequency , 1975 .

[28]  R. Rigler,et al.  Picosecond Fluorescence Spectroscopy in the Analysis of Structure and Motion of Biopolymers , 1985 .

[29]  A. Balter,et al.  A method of avoiding wavelength-dependent errors in decay-time measurements , 1979 .

[30]  R. D. Spencer,et al.  MEASUREMENTS OF SUBNANOSECOND FLUORESCENCE LIFETIMES WITH A CROSS‐CORRELATION PHASE FLUOROMETER * , 1969 .

[31]  R. E. Dale,et al.  Time-Resolved Fluorescence Spectroscopy in Biochemistry and Biology , 1983, NATO Advanced Science Institutes Series.

[32]  S. Canonica,et al.  Improved timing resolution using small side‐on photomultipliers in single photon counting , 1985 .

[33]  H. Haar,et al.  Phase fluorometer for measurement of picosecond processes. , 1978, The Review of scientific instruments.

[34]  Enrico Gratton,et al.  A multifrequency phase fluorometer using the harmonic content of a mode-locked laser , 1985 .

[35]  Graham R. Fleming,et al.  Direct observation of rotational diffusion by picosecond spectroscopy , 1976 .

[36]  Enrico Gratton,et al.  Multifrequency cross‐correlation phase fluorometer using synchrotron radiation , 1984 .

[37]  K. Berndt,et al.  Picosecond phase fluorometry by mode-locked cw lasers , 1982 .