Parallel confocal detection of single biomolecules using diffractive optics and integrated detector units
2004 (English)In: Current Pharmaceutical Biotechnology, ISSN 1389-2010, Vol. 5, no 2, 231-241 p.Article, review/survey (Refereed) Published
The past few years we have witnessed a tremendous surge of interest in so-called array-based miniaturised analytical systems due to their value as extremely powerful tools for high-throughput sequence analysis, drug discovery and development, and diagnostic tests in medicine (see articles in Issue 1). Terminologies that have been used to describe these array-based bioscience systems include (but are not limited to): DNA-chip, microarrays, microchip, biochip, DNA-microarrays and genome chip. Potential technological benefits of introducing these miniaturised analytical systems include improved accuracy, multiplexing, lower sample and reagent consumption, disposability, and decreased analysis times, just to mention a few examples. Among the many alternative principles of detection-analysis (e.g. chemiluminescence, electroluminescence and conductivity), fluo re scence-based techniques are widely used, examples being fluorescence resonance energy transfer, fluorescence quenching, fluorescence polarisation, time-resolved fluorescence, and fluorescence fluctuation spectroscopy (see articles in Issue II). Time-dependent fluctuations of fluorescent biomolecules with different molecular properties, like molecular weight, translational and rotational diffusion time, colour and lifetime, potentially provide all the kinetic and thermodynamic information required in analysing complex interactions. In this mini-review article, we present recent extensions aimed to implement parallel laser excitation and parallel fluorescence detection that can lead to even further increase in throughput in miniaturised array-based analytical systems. We also report on developments and characterisations of multiplexing extension that allow multifocal laser excitation together with matched parallel fluorescence detection for parallel confocal dynamical fluorescence fluctuation studies at the single biomolecule level.
Place, publisher, year, edition, pages
2004. Vol. 5, no 2, 231-241 p.
multiplexed excitation and detection, parallel confocal spectroscopy, high throughput screening, and fluorescence, fluctuation analysis, fluorescence cross-correlation, multifocal multiphoton microscopy, laser array sources, fan-out element, correlation spectroscopy, gene-expression, avalanche photodiodes, dna microarrays, tweezer arrays, genomics
IdentifiersURN: urn:nbn:se:kth:diva-23794ISI: 000224302600013OAI: oai:DiVA.org:kth-23794DiVA: diva2:342493
QC 201005252010-08-102010-08-10Bibliographically approved