Fluorescence intensity and anisotropy decays of the intrinsic tryptophan emission of hemoglobin measured with a 10-GHz fluorometer using front-face geometry on free liquid surface

Enrico Bucci, Zygmunt Gryczynski, Clara Fronticelli, Ignacy Gryczynski, Joseph R. Lakowicz

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

We measure the intensity and anisotropy decays of the intrinsic tryptophan emission from hemoglobin solutions obtained using a 10 GHz frequency-domain fluorometer and a specially designed cuvette which allows front face excitation on a free liquid surface. The cuvette eliminates reflections and stray emissions, which become significant for low intensity fluorescence like in hemoglobin. Three lifetimes are detectable in the subnanosecond range. The average lifetime of hemoglobin is ligand dependent. Fluorescence anisotropy decays of oxy, deoxy, and carbonmonoxyhemoglobin can be fitted with up to three correlation times. When three components are used the floating initial anisotropy ro is in each case higher than the steady-state anisotropy of tryptophan in vitrified solution. For deoxy hemoglobin it is close to 0.4. The data are consistent with an initial loss of anisotropy from 0.4 to about 0.2 occurring in the first two picoseconds.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
PublisherPubl by Int Soc for Optical Engineering
Pages556-561
Number of pages6
ISBN (Print)0819407860
StatePublished - Jan 1 1992
Externally publishedYes
EventTime-Resolved Laser Spectroscopy in Biochemistry III - Los Angeles, CA, USA
Duration: Jan 20 1992Jan 22 1992

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume1640
ISSN (Print)0277-786X

Other

OtherTime-Resolved Laser Spectroscopy in Biochemistry III
CityLos Angeles, CA, USA
Period1/20/921/22/92

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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