Michal Balberg | PubMed Health

PubMed2004

Microfluidic ELISA: on-chip fluorescence imaging.

Fluorescent reactions of a heterogeneous sandwich enzyme-linked immunoassay (ELISA) in an all-PDMS [poly (dimethylsiloxane)] microfluidic device were detected using a cooled charge coupled device (CCD) camera interfaced with an epifluorescence microscope. The study represents preliminary efforts to integrate biochemical reactions and detection on-chip using the "hybrid" detection approach. In initial experiments, the PDMS chip microsensor was successfully used to quantify a model analyte (sheep IgM) with sensitivity down to 17nM. Thus, we demonstrate here the extension of this hybrid integrated technique to on-chip imaging and quantification of light emission from a biochemical immunoassay in PDMS chip.

Animals↗

PubMed2003

Use of DNA and peptide nucleic acid molecular beacons for detection and quantification of rRNA in solution and in whole cells.

DNA and peptide nucleic acid (PNA) molecular beacons were successfully used to detect rRNA in solution. In addition, PNA molecular beacon hybridizations were found to be useful for the quantification of rRNA: hybridization signals increased in a linear fashion with the 16S rRNA concentrations used in this experiment (between 0.39 and 25 nM) in the presence of 50 nM PNA MB. DNA and PNA molecular beacons were successfully used to detect whole cells in fluorescence in situ hybridization (FISH) experiments without a wash step. The FISH results with the PNA molecular beacons were superior to those with the DNA molecular beacons: the hybridization kinetics were much faster, the signal-to-noise ratio was much higher, and the specificity was much better for the PNA molecular beacons. Finally, it was demonstrated that the combination of the use of PNA molecular beacons in FISH and flow cytometry makes it possible to rapidly collect quantitative FISH data. Thus, PNA molecular beacons might provide a solution for limitations of traditional FISH methods, such as variable target site accessibility, poor sensitivity for target cells with low rRNA content, background fluorescence, and applications of FISH in microfluidic devices.

Base Sequence↗