Development of a Multichannel Fluorescence Affinity Sensor System

Abstract

A multichannel fluorometer is proposed for analysis of biochemical reactions. The sensor is based on the luminescence generation in the evanescent field of a totally reflected laser beam. For transduction, multiple reflection elements are used. Multichannel operation is realized, including the possibility of applying different solutions to each channel at the same time. First experimental results, obtained with fluorescein or Cy5 as labels in a model hybridization assay, demonstrate the applicability and allow the detection of 3-10 fmol injected fluorescently labeled oligonucleotide.

Optical microarray biosensing techniques

“…81 The sensitivity was found to be similar to standard enzymelinked immunosorbent assay (ELISA) test assays. 85,88 Bulk waveguide TIRF setups for the detection of DNA hybridization have also been reported, such as the multichannel sensor presented by Schuderer et al 91 and the microarray format using standard microscopy slides described by Lehr et al 92 For these applications, DNA was immobilized either by interaction of biotinylated oligonucleotides with a glass slide coated with avidin and backfilled with bovine serum albumin (BSA) 91 or by spotting plastic slides or epoxy-functionalized glass slides with oligonucleotides and subsequent crosslinking. 92 In addition, ASPER Biotech (Tartu, Estonia) has developed a commercially available microarray system, based on multiple TIRF, for medium-throughput genotyping.…”

Trends and Perspectives in Immunosensors

“…It has a maximum intensity at the surface, which is reduced with increasing distance from the surface [92]. These sensors demonstrate high sensitivity and can avoid separation and extraction steps from complex matrices such as body fluids [93,94]. As an example, studies were carried out using TIRF immunosensors to evaluate the concentration of bovine progesterone, a reproductive hormone present in raw milk [95,96].…”

TIRF Array Biosensor for Environmental Monitoring

“…The most commonly used lasers are the argon-ion ( 488 nm) laser for fluorescein and a helium-neon (633 nm) or diode laser (635 nm) for the cyanine dye (Cy5). A number of devices have been used in the detection of the resulting fluorescence emission, in particular CCD cameras [70][71][72], photomultiplier tubes (PMT) [73], photodiodes [74], a single photomultiplier tube [75,76] and more recently a complementary metal oxide-semiconductor (CMOS) camera [77].…”

“…There is currently only one example of measuring DNA or mRNA hybridization between complementary strands using interferometry [34]; however, the process has become increasingly studied using planar waveguide TIRF [76,81,88]. Duveneck et al [81] coupled 16-mer oligonucleotides to functionalized tantalum pentoxide waveguides and studied the specific binding of Cy5-labeled complementary oligonucleotides using TIRF.…”