Biodiagnostics and Polymer Identification with Multiplex Dyes

Abstract

A new analytical concept based on time‐resolved fluorescence spectroscopy is presented, which uses the characteristic fluorescence lifetimes as information to identify new fluorescent dyes, called multiplex dyes. Several dyes can be distinguished at nearly fixed excitation and emission wavelength by recognition of the lifetimes. A pattern recognition technique is used for data analysis. As light sources a pulsed laser diode is used. First experiments are shown in several application fields, like DNA sequencing, immunoassays and tagging of polymer material.

Near‐infrared time‐resolved fluorescence lifetime determinations in poly(methylmethacrylate) microchip electrophoresis devices

“…Coupling fluorescence lifetime detection with LIF can produce steady-state data, but allows the ability to monitor several fluorophores possessing distinguishable lifetime values simultaneously for highly multiplexed assays [7]. One of the advantages of fluorescence lifetime determinations is that the lifetime is independent of concentration and the excitation wavelength [8].…”

Simultaneous antigen detection using multiplex dyes

“…In addition to the spectrum, the lifetime of the excited state is a characteristic feature of fluorescent dyes. Introducing the fluorescence lifetime as a characteristic parameter of dyes, the concept of simultaneous detection can be extended [4]. Hence, fluorescent dyes whose absorption and emission spectra are nearly identical, but which differ in fluorescence lifetime have to be designed [4,51. These so-called "multiplex dyes" can be linked to antibodies or other biomolecules by in situ activated succinimido esters of the dyes, which react with the amino functions of the basic lysine residues.…”

Alkylene-bridged N,N,N′N′-Tetrasubstituted Bis(2-amino-5-thiazolyl)methinium Salts-A New Class of Strongly Fluorescent Dyes

“…Owing to their strong fluorescence, pyronines 2 a (R 1 =H)1 and rhodamines 2 a (R 1 =Aryl)2 have found several practical applications, such as fluorescence and laser dyes,3 or, if they are specifically functionalized, for example at the amino groups, as fluorescence markers for biological substrates and polymer characterization 4. The fluorescence of the dyes 2 a is in sharp contrast to their unbridged di‐ and triarylmethane dye analogues 1 which are nonfluorescent under normal conditions 5.…”