Ultrasensitive fluorescence correlation spectroscopy of highly parallelized microfluidic devices

Brian K. Canfield; Jason K. King; William N. Robinson; William H. Hofmeister; Steven A. Soper; Lloyd M. Davis

doi:10.1117/12.909515

13 February 2012 Ultrasensitive fluorescence correlation spectroscopy of highly parallelized microfluidic devices

Brian K. Canfield, Jason K. King, William N. Robinson, William H. Hofmeister, Steven A. Soper, Lloyd M. Davis

Proceedings Volume 8228, Single Molecule Spectroscopy and Superresolution Imaging V; 82280G (2012) https://doi.org/10.1117/12.909515
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

Reducing reagent needs and costs while increasing throughput constitute important needs for assays in pharmaceutical drug discovery. We are developing an ultrasensitive, fluorescence-based detection system in highly parallel microfluidic channels with kHz readout rates in each channel. Prototype microfluidic devices with an array of >150 microchannels have been fabricated by direct femtosecond laser machining of fused silica substrates. A device is placed in a custombuilt, wide-field microscope where a line-generating red diode laser provides uniform epi-illumination just a few microns high across a 500 micron field of view. Single-molecule levels in the probe volumes can be attained by flowing suitably dilute aqueous solutions (~pM) of fluorescently labeled biomolecules through the microchannels. Fluorescence is detected with an electron-multiplying CCD camera allowing readout rates up to 7 kHz for each microchannel. Rapid initial assessment of detected fluorescence signals is performed through digital filtering derived from simulations based on experimental parameters. Fluorescence correlation spectroscopy can then provide more detailed analysis of the sample within each microchannel. Optimized microfluidic devices could be mass-produced in low-cost polymers using imprint lithography.

Citation Download Citation

Brian K. Canfield, Jason K. King, William N. Robinson, William H. Hofmeister, Steven A. Soper, and Lloyd M. Davis "Ultrasensitive fluorescence correlation spectroscopy of highly parallelized microfluidic devices", Proc. SPIE 8228, Single Molecule Spectroscopy and Superresolution Imaging V, 82280G (13 February 2012); https://doi.org/10.1117/12.909515

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KEYWORDS

Microfluidics

Signal detection

Luminescence

Silica

Microscopes

Digital filtering

Femtosecond phenomena

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