A bead-based fluorescence immunosensing technique enabled by the integration of Förster resonance energy transfer and optoelectrokinetic concentration

Abstract

Bead-based immunosensing has been growing as a promising technology in the point-of-care diagnostics due to great flexibility. For dilute samples, functionalized particles can be used to collect dispersed analytes and act as carriers for particle manipulation. To realize rapid and visual immunosensing, F€ orster resonance energy transfer (FRET) was used herein to ensure only the diabetic biomarker, lipocalin 1, to be detected. The measurement was made in an aqueous droplet sandwiched between two parallel plate electrodes. With an electric field and a focused laser beam applying on the microchip simultaneously, the immunocomplexes in the droplet were further concentrated to enhance the FRET fluorescent signal. The optoelectrokinetic technique, termed rapid electrokinetic patterning (REP), has been proven to be excellent in dynamic and programmable particle manipulation. Therefore, the detection can be complete within several tens of seconds. The lower detection limit of the REP-enabled bead-based diagnosis reached nearly 5 nM. The combinative use of FRET and the optoelectrokinetic technique for the bead-based immunosensing enables a rapid measure to diagnose early stage diseases and dilute analytes. V C 2016 AIP Publishing LLC. [http://dx

Bead based facile assay for sensitive quantification of native state green fluorescent protein

“…This is useful method for complex proteomics studies and biomedical studies to identify potential target protein for various applications. [31][32][33][34]…”

Sensitive tear screening of diabetic retinopathy with dual biomarkers enabled using a rapid electrokinetic patterning platform

“…The fluorescence intensity of the microbeads was enhanced as they are concentrated by REP. We previously showed that microbead manipulation is a function of electrical frequency. 20,24,34 In this study, the optimal manipulation of 3 μm microbeads by REP was realized with 15 V pp at 35 kHz and 12 mW laser intensity. The fluorescence of the microbeads dynamically aggregated under the operating conditions was excited and emitted through a filter cube equipped in the microscope.…”

“…In the present study, a non-invasive immunosensing technique ( Fig. 1) based on an optoelectrokinetic platform, termed rapid electrokinetic patterning (REP), [19][20][21][22][23][24] was used to manipulate immunocomplexed microbeads for the detection of DR biomarkers in tear samples. We utilized a colloidal suspension containing microbeads conjugated with capture and free suspending fluorescence tagged probe antibodies.…”

A simple electrokinetic protein preconcentrator utilizing nano-interstices

“…Particle concentration had been done adopting the optoelectrokinetic technique, termed rapid electrokinetic patterning (REP). 1,2 With an electric field and a focused laser beam applying on the microchip simultaneously, the immunocomplexes in the droplet of sample were further concentrated to enhance the F€ orster resonance energy transfer (FRET) fluorescent signal. 2 This approach is relative complicated and the rising of temperature might damage the analytes.…”