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Proceedings Paper

Combining surface sensitive vibrational spectroscopy and fluorescence microscopy to study biological interfaces
Author(s): Chi Zhang; Joshua Jasensky; Jing Wu; Zhan Chen
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Paper Abstract

A multimodal system combining surface sensitive sum frequency generation (SFG) vibrational spectroscopy and total-internal reflection fluorescence (TIRF) microscopy for surface and interface study was developed. Interfacial molecular structural information can be detected using SFG spectroscopy while interfacial fluorescence signal can be visualized using TIRF microscopy from the same sample. As a proof of concept experiment, SFG spectra of fluorescent polystyrene (PS) beads with different surface coverage were correlated with TIRF signal observed. Results showed that SFG signals from the ordered surfactant methyl groups were detected from the substrate surface, while signals from PS phenyl groups on the beads were not seen. Additionally, a lipid monolayer labeled using lipid-associated dye was deposited on a silica substrate and studied in different environments. The contact with water of this lipid monolayer caused SFG signal to disappear, indicating a possible lipid molecular disorder and the formation of lipid bilayers or liposomes in water. TIRF was able to visualize the presence of lipid molecules on the substrate, showing that the lipids were not removed from the substrate surface by water. The integration of the two surface sensitive techniques can simultaneously visualize interfacial molecular dynamics and characterize interfacial molecular structures in situ, which is important and is expected to find extensive applications in biological interface related research.

Paper Details

Date Published: 10 April 2014
PDF: 8 pages
Proc. SPIE 8947, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XII, 894712 (10 April 2014); doi: 10.1117/12.2040828
Show Author Affiliations
Chi Zhang, Univ. of Michigan (United States)
Joshua Jasensky, Univ. of Michigan (United States)
Jing Wu, Univ. of Michigan (United States)
Zhan Chen, Univ. of Michigan (United States)


Published in SPIE Proceedings Vol. 8947:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XII
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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