Ultrasensitive near-IR fluorescence detection in capillary zone electrophoresis

Steven A. Soper; Benjamin L. Legendre Jr.; James H. Flanagan Jr.; Daryl C. Williams; Robert P. Hammer

doi:10.1117/12.180791

21 July 1994 Ultrasensitive near-IR fluorescence detection in capillary zone electrophoresis

Steven A. Soper, Benjamin L. Legendre Jr., James H. Flanagan Jr., Daryl C. Williams, Robert P. Hammer

Proceedings Volume 2136, Biochemical Diagnostic Instrumentation; (1994) https://doi.org/10.1117/12.180791
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

Capillary zone electrophoresis (CZE) is a powerful new separation technique which possesses the ability to separate small and large molecules. Due to the small volume of material that is typically loaded onto the column, ultrasensitive detection is often required. We have constructed a laser-induced fluorescence detector appropriate for CZE applications using near- IR fluorescence excitation and detection. Separations of native NIR fluorescent dyes in nonaqueous running buffer systems have been performed with detection sensitivities in the low zeptomole range, comparable to state-of-the-art detection limits reported for visible fluorescence detection in CZE. Using our NIR fluorescence detector and NIR labeling dyes synthesized in our laboratory, we present electropherograms of peptides and proteins separated by CZE and detected with NIR fluorescence. We also discuss the basic operating principles of CZE as well as the instrumental components necessary to perform CZE separations.

Citation Download Citation

Steven A. Soper, Benjamin L. Legendre Jr., James H. Flanagan Jr., Daryl C. Williams, and Robert P. Hammer "Ultrasensitive near-IR fluorescence detection in capillary zone electrophoresis", Proc. SPIE 2136, Biochemical Diagnostic Instrumentation, (21 July 1994); https://doi.org/10.1117/12.180791

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