Frequency-domain fluorescence spectroscopy: instrumentation and applications to the biosciences (Keynote Paper)

Joseph R. Lakowicz; Ignacy Gryczynski; Henryk Malak; Michael L. Johnson; Gabor Laczko; Wieslaw M. Wiczk; Henryk Szmacinski; Jozef Kusba

Frequency-domain fluorescence spectroscopy: instrumentation and applications to the biosciences (Keynote Paper)

Joseph R. Lakowicz, Ignacy Gryczynski, Henryk Malak, Michael L. Johnson, Gabor Laczko, Wieslaw M. Wiczk, Henryk Szmacinski, Jozef Kusba

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

1 Scopus citations

Abstract

Measurements of time-resolved fluorescence are increasingly used for research in biophysics, biochemistry, cell biology and medicine. Advances in the technology of light sources and detectors are resulting in more reliable and/or advanced instrumentation, which is resulting in the expanding applications of fluorescence spectroscopy. Time-resolved measurements are often performed by direct measurements in the time-domain. In this article the authors describe the alternative method of frequency-domain fluorometry. The frequency-response of the emission to intensity-modulated excitation can be used to recover the time-dependent decay. Commercial instrumentation now allows measurements to an upper light modulation frequency limit of 200 MHz. This laboratory has developed second and third generation instruments which allows measurements to 2 GHz and subsequently to 10 GHz. The frequency-domain data from such instrumentation provides excellent resolution of picosecond decays of intensity and anisotropy. Additionally, the frequency-domain method appears to provide remarkable resolution of complex decays which are often observed for biochemical samples. In this article the authors describe this instrumentation and applications of this method. Examples are shown using probes with ps decay and correlation times, the intrinsic fluorescence of proteins, and the measurement of end-to-end diffusion in proteins and/or flexible molecules.

Original language	English
Title of host publication	Proceedings of SPIE - The International Society for Optical Engineering
Publisher	Publ by Int Soc for Optical Engineering
Pages	142-160
Number of pages	19
ISBN (Print)	0819405256
State	Published - 1991
Event	Optical Methods for Ultrasensitive Detection and Analysis: Techniques and Applications - Los Angeles, CA, USA Duration: 21 Jan 1991 → 23 Jan 1991

Publication series

Name	Proceedings of SPIE - The International Society for Optical Engineering
Volume	1435
ISSN (Print)	0277-786X

Other

Other	Optical Methods for Ultrasensitive Detection and Analysis: Techniques and Applications
City	Los Angeles, CA, USA
Period	21/01/91 → 23/01/91

Cite this

Lakowicz, J. R., Gryczynski, I., Malak, H., Johnson, M. L., Laczko, G., Wiczk, W. M., Szmacinski, H., & Kusba, J. (1991). Frequency-domain fluorescence spectroscopy: instrumentation and applications to the biosciences (Keynote Paper). In Proceedings of SPIE - The International Society for Optical Engineering (pp. 142-160). (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 1435). Publ by Int Soc for Optical Engineering.

Lakowicz, Joseph R. ; Gryczynski, Ignacy ; Malak, Henryk et al. / Frequency-domain fluorescence spectroscopy : instrumentation and applications to the biosciences (Keynote Paper). Proceedings of SPIE - The International Society for Optical Engineering. Publ by Int Soc for Optical Engineering, 1991. pp. 142-160 (Proceedings of SPIE - The International Society for Optical Engineering).

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title = "Frequency-domain fluorescence spectroscopy: instrumentation and applications to the biosciences (Keynote Paper)",

abstract = "Measurements of time-resolved fluorescence are increasingly used for research in biophysics, biochemistry, cell biology and medicine. Advances in the technology of light sources and detectors are resulting in more reliable and/or advanced instrumentation, which is resulting in the expanding applications of fluorescence spectroscopy. Time-resolved measurements are often performed by direct measurements in the time-domain. In this article the authors describe the alternative method of frequency-domain fluorometry. The frequency-response of the emission to intensity-modulated excitation can be used to recover the time-dependent decay. Commercial instrumentation now allows measurements to an upper light modulation frequency limit of 200 MHz. This laboratory has developed second and third generation instruments which allows measurements to 2 GHz and subsequently to 10 GHz. The frequency-domain data from such instrumentation provides excellent resolution of picosecond decays of intensity and anisotropy. Additionally, the frequency-domain method appears to provide remarkable resolution of complex decays which are often observed for biochemical samples. In this article the authors describe this instrumentation and applications of this method. Examples are shown using probes with ps decay and correlation times, the intrinsic fluorescence of proteins, and the measurement of end-to-end diffusion in proteins and/or flexible molecules.",

author = "Lakowicz, {Joseph R.} and Ignacy Gryczynski and Henryk Malak and Johnson, {Michael L.} and Gabor Laczko and Wiczk, {Wieslaw M.} and Henryk Szmacinski and Jozef Kusba",

year = "1991",

language = "English",

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series = "Proceedings of SPIE - The International Society for Optical Engineering",

publisher = "Publ by Int Soc for Optical Engineering",

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Lakowicz, JR, Gryczynski, I, Malak, H, Johnson, ML, Laczko, G, Wiczk, WM, Szmacinski, H & Kusba, J 1991, Frequency-domain fluorescence spectroscopy: instrumentation and applications to the biosciences (Keynote Paper). in Proceedings of SPIE - The International Society for Optical Engineering. Proceedings of SPIE - The International Society for Optical Engineering, vol. 1435, Publ by Int Soc for Optical Engineering, pp. 142-160, Optical Methods for Ultrasensitive Detection and Analysis: Techniques and Applications, Los Angeles, CA, USA, 21/01/91.

Frequency-domain fluorescence spectroscopy : instrumentation and applications to the biosciences (Keynote Paper). / Lakowicz, Joseph R.; Gryczynski, Ignacy; Malak, Henryk et al.

Proceedings of SPIE - The International Society for Optical Engineering. Publ by Int Soc for Optical Engineering, 1991. p. 142-160 (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 1435).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › peer-review

TY - GEN

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AU - Gryczynski, Ignacy

AU - Malak, Henryk

AU - Johnson, Michael L.

AU - Laczko, Gabor

AU - Wiczk, Wieslaw M.

AU - Szmacinski, Henryk

AU - Kusba, Jozef

PY - 1991

Y1 - 1991

N2 - Measurements of time-resolved fluorescence are increasingly used for research in biophysics, biochemistry, cell biology and medicine. Advances in the technology of light sources and detectors are resulting in more reliable and/or advanced instrumentation, which is resulting in the expanding applications of fluorescence spectroscopy. Time-resolved measurements are often performed by direct measurements in the time-domain. In this article the authors describe the alternative method of frequency-domain fluorometry. The frequency-response of the emission to intensity-modulated excitation can be used to recover the time-dependent decay. Commercial instrumentation now allows measurements to an upper light modulation frequency limit of 200 MHz. This laboratory has developed second and third generation instruments which allows measurements to 2 GHz and subsequently to 10 GHz. The frequency-domain data from such instrumentation provides excellent resolution of picosecond decays of intensity and anisotropy. Additionally, the frequency-domain method appears to provide remarkable resolution of complex decays which are often observed for biochemical samples. In this article the authors describe this instrumentation and applications of this method. Examples are shown using probes with ps decay and correlation times, the intrinsic fluorescence of proteins, and the measurement of end-to-end diffusion in proteins and/or flexible molecules.

AB - Measurements of time-resolved fluorescence are increasingly used for research in biophysics, biochemistry, cell biology and medicine. Advances in the technology of light sources and detectors are resulting in more reliable and/or advanced instrumentation, which is resulting in the expanding applications of fluorescence spectroscopy. Time-resolved measurements are often performed by direct measurements in the time-domain. In this article the authors describe the alternative method of frequency-domain fluorometry. The frequency-response of the emission to intensity-modulated excitation can be used to recover the time-dependent decay. Commercial instrumentation now allows measurements to an upper light modulation frequency limit of 200 MHz. This laboratory has developed second and third generation instruments which allows measurements to 2 GHz and subsequently to 10 GHz. The frequency-domain data from such instrumentation provides excellent resolution of picosecond decays of intensity and anisotropy. Additionally, the frequency-domain method appears to provide remarkable resolution of complex decays which are often observed for biochemical samples. In this article the authors describe this instrumentation and applications of this method. Examples are shown using probes with ps decay and correlation times, the intrinsic fluorescence of proteins, and the measurement of end-to-end diffusion in proteins and/or flexible molecules.

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M3 - Conference contribution

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SN - 0819405256

T3 - Proceedings of SPIE - The International Society for Optical Engineering

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BT - Proceedings of SPIE - The International Society for Optical Engineering

PB - Publ by Int Soc for Optical Engineering

Y2 - 21 January 1991 through 23 January 1991

ER -

Lakowicz JR, Gryczynski I, Malak H, Johnson ML, Laczko G, Wiczk WM et al. Frequency-domain fluorescence spectroscopy: instrumentation and applications to the biosciences (Keynote Paper). In Proceedings of SPIE - The International Society for Optical Engineering. Publ by Int Soc for Optical Engineering. 1991. p. 142-160. (Proceedings of SPIE - The International Society for Optical Engineering).

Frequency-domain fluorescence spectroscopy: instrumentation and applications to the biosciences (Keynote Paper)

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