Design of a fluorescence-lifetime imaging microscope workstation
One of the promising recent developments in fluorescence microscopy is fluorescence lifetime imaging microscopy. This type of microscopy images the lifetime of fluorescence molecules (in the nano seco...
Single-photon timing fluorometry using three-photon excitation
We demonstrate the application of three photon excitation to fluorescence probe studies using time-correlated single- photon counting. By exciting with 120 fs Ti:sapphire laser pulses at 800 nm we hav...
Proc. SPIE, Vol. 2980, 42 (1997);
doi:10.1117/12.273565
Online Publication Date: 8 April 2005
Conference Date: Sunday 09 February 1997
Conference Location: San Jose, CA, USA
Conference Title: Advances in Fluorescence Sensing Technology III
Conference Chairs: Richard B. Thompson
ABSTRACT
Lia A. Avanessian Institute of Epidemiology, Virology and Medical Parisitology (Armenia)
Preliminaryresults of the investigation of two-photon excited fluorescence of acridineorange (AO) and ethidium bromide (EB) in complexes with DNAare presented. Spectrofluorometer based on picosecond Nd:YAG laser was usedfor investigations on two-photon (1064 nm, 1 mJ, 40 ps)and one-photon (532 and 355 nm) excitation. Highly purified DNAfrom beef spleen and AO, EB were used at theconcentration of: AO- 15 (mu) M, EB- 12 (mu) M,DNA- 2-15 (mu) g/ml. The spectra of two-photon excited fluorescenceof AO, EB and their complexes with DNA as wellas kinetics of the intensification of dye's two-photon fluorescence duringtheir interactions with DNA in dependence on DNA concentration wereobtained. The intensity of fluorescence of AO and EB wasincreased maximum 2.4 and 8 times correspondingly. During the one-photonexcitation corresponding values were for: AO-2.5 time and EB-10 time.The difference in the intensification of EB fluorescence connected withthe difference of the change of one-photon and two-photon absorptioncoefficients during the interaction of the dye with DNA.
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