Principles of FRET – Based (Bio) Sensors

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Presentation transcript:

Principles of FRET – Based (Bio) Sensors Nanoscience Symposium 2005 Auke Jisk Kronemeijer

Introduction Need for sensitive, reusable measurement devices Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Introduction Need for sensitive, reusable measurement devices Specific molecular detection Specific detection of activities Towards in vivo detection

Interesting Principle For Sensors is FRET Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Interesting Principle For Sensors is FRET Fluorescence / Förster Resonance Energy Transfer Why FRET ? -Distance and Spectral Overlap dependences Possibility for Processing in Integrated Circuits

General Principles of Resonance Energy Transfer Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 General Principles of Resonance Energy Transfer - Radiative vs non-radiative - Two models for non-radiative Förster Dexter

General Principles of Resonance Energy Transfer Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 General Principles of Resonance Energy Transfer - Detection of FRET - Quantitative Detection Difficulties Spectral Bleed-Through Acceptor excitation by donor excitation wavelenght Cross Talk Donor Emission Contribution to FRET Emission

FRET Based Recognition Biosensor Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 FRET Based Recognition Biosensor - Modified Biomolecule for Specificity

Problems with Molecular Principle Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Problems with Molecular Principle Large spectral bleed-through and cross-talk Difficult to find good / compatible pairs

Solutions to Spectral Bleed-Through Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Solutions to Spectral Bleed-Through Photochromic FRET “Switching on and off FRET mechanism to determine FRET contribution to total emission”

Solutions to Spectral Bleed-Through Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Solutions to Spectral Bleed-Through Use Quantum Dots as Donors Advantages: Broad absorption Narrow Emission Size tunability Surface functionalisation

Quantum Dot FRET Based Recognition Sensor Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Quantum Dot FRET Based Recognition Sensor Possibility for Templating AFM Image of Si8Ge2 QD´s on Si(100)

Sensors Based on Distance Changes Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Sensors Based on Distance Changes Enzyme Activity Sensors Reminder:

Recognition Sensor based on Isomerisation Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Sensors Based on Distance Changes Determination of Isomerisation Recognition Sensor based on Isomerisation If Initiator / Target is Present, FRET Efficiency Changes

Conclusions Many different sensors possible with use of FRET Principles of FRET – Based (Bio) Sensors - Nanoscience Symposium 2005 Conclusions Many different sensors possible with use of FRET Use of Quantum Dots as Donor Promising Innovative Solutions Come Up for Quantitative Measurements