A new front-face optical cell for measuring weak fluorescent emissions with time resolution in the picosecond time scale.
Recent developments of ultrafast fluorimeters allow measuring time-resolved fluorescence on the picosecond time scale. This implies one is able to monitor lifetimes and anisotropy decays of highly quenched systems and of systems that contain fluorophores having lifetimes in the subnanosecond range; both systems that emit weak signals. The combination of weak signals and very short lifetimes makes the measurements prone to distortions which are negligible in standard fluorescence experiments. To cope with these difficulties, we have designed a new optical cell for front-face optics which offers to the excitation beam a horizontal free liquid surface in the absence of interactions with optical windows. The new cell has been tested with probes of known lifetimes and anisotropies. It proved very useful in detecting tryptophan fluorescence in hemoglobin. If only diluted samples are available, which cannot be used in front-face optics, regular square geometry can still be utilized by inserting light absorbers into a cuvette of 1 cm path length.
Gryczynski
Z
Z
Department of Biochemistry, University of Maryland at Baltimore 21201.
Bucci
E
E
D002623:Chemistry Techniques, Analytical / Q000295:instrumentation* / Q000379:methods; D005454:Fluorescence Polarization; D055095:Optics and Photonics / Q000295:instrumentation; D010108:Oxyhemoglobins / Q000737:chemistry; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods; D013059:Spectrum Analysis, Raman; D013997:Time Factors; D014364:Tryptophan / Q000737:chemistry
D010108:Oxyhemoglobins; D014364:Tryptophan