DOI: 10.1063/1.1137078. Modulation detection of fluorescence photobleaching recovery

Modulation detection of fluorescence photobleaching recovery

10.1063/1.1137078

Crossref journal-article

AIP Publishing

Review of Scientific Instruments (317)

Abstract

We have combined the simplicity of the periodic pattern method of photobleaching with the signal processing advantage of scanning detection. A periodic photobleaching pattern is produced in the sample by brief, intense illumination through a grating. The grating is then translated at constant speed through the reference beam to produce a moving pattern on the specimen. A modulation of fluorescence emission is produced as the bleached pattern and the illumination pattern fall into and out of phase. The resulting photocurrent contains an ac component whose frequency is determined by the spacing and velocity of the grating and whose amplitude relative to the dc component is determined by the extent of photobleaching. The decay constant of the ac component of the photocurrent is determined by the spacing of the grating and the diffusion coefficient of the labeled species. The major distinct advantages of the technique are: (1) the decay at the fundamental frequency is observed in real time, (2) adventitious photobleaching during the monitoring phase of the measurement does not perturb the determined parameters, (3) the measurement is insensitive to dc drift components, and (4) the measurement is insensitive to motion of the specimen. These substantial advantages are implemented by straightforward modifications in standard methodology.

Bibliography

Lanni, F., & Ware, B. R. (1982). Modulation detection of fluorescence photobleaching recovery. Review of Scientific Instruments, 53(6), 905â908.

Authors 2

Frederick Lanni (first)
B. R. Ware (additional)

References 11 Referenced 70

10.1016/0005-2736(74)90085-6 / Biochim. Biophys. Acta (1974)
10.1016/0005-2736(76)90189-9 / Biochim. Biophys. Acta (1976)
10.1016/S0006-3495(76)85755-4 / Biophys. J. (1976)
10.1016/S0006-3495(76)85776-1 / Biophys. J. (1976)
10.1073/pnas.75.6.2759 / Proc. Natl. Acad. Sci. USA (1978)
10.1073/pnas.76.11.5641 / Proc. Natl. Acad. Sci. USA (1979)
10.1073/pnas.76.7.3314 / Proc. Natl. Acad. Sci. USA (1979)
10.1016/S0006-3495(81)84794-7 / Biophys. J. (1981)
10.1016/S0006-3495(79)85176-0 / Biophys. J. (1979)
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Dates

Type	When
Created	22 years, 5 months ago (March 10, 2003, 1:11 p.m.)
Deposited	1 year, 6 months ago (Feb. 6, 2024, 1:43 a.m.)
Indexed	1 year ago (Sept. 2, 2024, 2:28 a.m.)
Issued	43 years, 3 months ago (June 1, 1982)
Published	43 years, 3 months ago (June 1, 1982)
Published Print	43 years, 3 months ago (June 1, 1982)

Funders 0

None

BibTeX

@article{Lanni_1982, title={Modulation detection of fluorescence photobleaching recovery}, volume={53}, ISSN={1089-7623}, url={http://dx.doi.org/10.1063/1.1137078}, DOI={10.1063/1.1137078}, number={6}, journal={Review of Scientific Instruments}, publisher={AIP Publishing}, author={Lanni, Frederick and Ware, B. R.}, year={1982}, month=jun, pages={905–908} }

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