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Single-particle identification of encoded nanospheres
Hendrik Hippchen1, Wiebke H Pohl, Peter J Walla
1Max Planck Institute for Biophysical Chemistry, Biomolecular Spectroscopy and Single-Molecule Detection, Göttingen, Germany.
Journal of Biomolecular Screening
|January 21, 2010
Summary
This study introduces a method to identify differently encoded nanospheres using fluorescence. This enables simultaneous measurement of multiple biomolecular interactions in a single assay.
Area of Science:
- Biophysics
- Analytical Chemistry
- Biochemistry
Background:
- Glycosylated nanospheres mimic cell surfaces for studying protein-carbohydrate interactions.
- Analyzing mixtures requires distinguishing individual nanospheres based on fluorescent labels.
Purpose of the Study:
- To develop a method for accurate, single-particle identification of differentially encoded nanospheres.
- To enable high-throughput analysis of biomolecular interactions in a complex environment.
Main Methods:
- Utilized 2-photon fluorescence correlation spectroscopy.
- Developed a technique for identifying encoded nanospheres during single transits through a microscope's focal volume.
- Employed differentially fluorescently labeled nanospheres in aqueous solution.
Main Results:
- Achieved nearly 100% accuracy in identifying individual encoded nanospheres.
- Demonstrated the ability to distinguish nanospheres with different fluorescent labels.
- Validated the identification method for nanospheres in aqueous solution.
Conclusions:
- The presented method allows for the certain identification of encoded nanospheres.
- This facilitates multiplexed, 1-well measurements of biomolecular receptor-ligand interactions.
- Enables testing of ligand behavior in mimicked complex environments and rapid screening of interactions.
