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Correction of fluorescence detected circular dichroic photoselection artifacts using light collecting ellipsoidal mirrors

Journal Article · · Rev. Sci. Instrum.; (United States)
DOI:https://doi.org/10.1063/1.1139463· OSTI ID:5984767

Ellipsoidal mirrors are used to collect the emitted light from fluorophores in fluorescence detected circular dichroic (FDCD) measurements and to deliver this light to a photomultiplier tube (PMT) at 90/sup 0/ to the excitation beam. These mirrors collect light over a symmetrical 71% of the emission sphere centered on the sample cell, and thereby eliminate most of the nonuniformity in detected emission resulting from the anisotropy of photoselection. Furthermore, even with reflectance and path length losses, the amount of fluorescent emission reaching the PMT is increased by a factor of approximately 18. Elimination of photoselection artifacts in UV FDCD spectra is demonstrated by comparing spectra of L-tryptophan in glycerol with its spectra in water. Validity of the values measured is verified through calibration with a d10 camphorsulfonic acid (d10 CSA) standard.

Research Organization:
Department of Biophysics, University of California, Berkeley, California 94720
OSTI ID:
5984767
Journal Information:
Rev. Sci. Instrum.; (United States), Journal Name: Rev. Sci. Instrum.; (United States) Vol. 58:11; ISSN RSINA
Country of Publication:
United States
Language:
English