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Surface-bound molecular beacons (MBs) were optimized for biosensing using surface plasmon fluorescence spectroscopy. Longer MBs enhanced fluorescence intensity for sensitive picomolar detection of target molecules.

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Analytical Chemistry

Background:

  • Surface-bound molecular beacons (MBs) are crucial for developing sensitive biosensors.
  • Optimizing MB surface immobilization and structure is key to enhancing optical readout signals.
  • Surface plasmon fluorescence spectroscopy (SPFS) offers a sensitive platform for analyzing surface-bound molecules.

Purpose of the Study:

  • To investigate the impact of molecular beacon (MB) length, linker, and surface chemistry on SPFS performance.
  • To optimize sensor chip preparation for sensitive and selective target molecule detection.
  • To evaluate the reusability and stability of MB-modified sensor chips.

Main Methods:

  • Chemisorption of thiol-functionalized MBs onto gold surfaces with varying thiol spacers.
  • Utilizing surface plasmon fluorescence spectroscopy (SPFS) for optical readout.
  • Characterizing MB distribution using fluorescence microscopy.

Main Results:

  • MB length and surface orientation significantly influenced fluorescence intensity after target hybridization.
  • Optimal sensor design achieved a limit of detection in the high picomolar range with response times of 5-20 minutes.
  • The system demonstrated selectivity for fully complementary targets over mismatched ones.
  • Sensor chips exhibited repeatable preparation and reusability without performance degradation.

Conclusions:

  • Surface-bound MBs, when optimized for length and surface attachment, provide a robust platform for sensitive biosensing.
  • SPFS is an effective readout technique for evaluating MB-based sensor performance.
  • The developed sensor chips offer a repeatable, reusable, and selective method for target molecule detection.