Make it Rigid

DESCRIPTION

This study explores a novel application of the Stark effect for calcium sensing in a Total Internal Reflection Fluorescence (TIRF) microscope setup. The focus is on optimizing TIRF configurations, including the use of specific objectives and prisms, to enhance fluorescence sensitivity. Key challenges addressed include managing background noise from excitation light, thermal interference, and shot noise, which can obscure the signal from individual dye molecules. A new concept of absorption cross-section is introduced to improve detection and measurement accuracy in membrane voltage sensing applications.

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May 17, 2026

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Make it Rigid

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Make it Rigid Ca++ Application: Calcium sensing
Stark effect Application: Membrane Voltage Sensing
TIRF Microscope

Setting up the TIRF microscope Objective-TIRF Prism-TIRF
Sensitivity of Fluorescence Microscope • Signal • New concept: absorption cross-section • Background due to excitation light: ~105 x (signal from one dye molecule) • Noise • Shot noise (unavoidable) • Thermal background noise • Read noise