Abstract

Measurements of cellular pH are used to infer information such as stage of cell cycle, presence of cancer and other diseases, as well as delivery or effect of a therapeutic drug. Surface-enhanced Raman spectroscopy (SERS) of nanoparticle-based pH probes have been used to interrogate intracellular pH, with the significant advantage of avoiding photobleaching compared to fluorescent indicators. 4-Mercaptobenzoic acid (MBA) is a commonly used pH-sensitive reporter molecule. Intracellular pH sensing by SERS requires analysis of the observed MBA spectrum and spectral interference can affect the pH determination. Background from common cell containers, imaging too few particles, signal-to-noise ratios, and degradation of reporter molecules are among the factors that may alter appropriate SERS-based pH determination in cells. Here, we have compared common methods of spectral analysis to see how different factors alter the calculated pH in Raman maps of MBA functionalized Au nanostars in SW620 cancer cells. The methods included in our comparison use the relative intensity of the ν(COO) stretch, chemometric analysis of the ν8a mode, and analyzing the frequency shift of the ν8a mode. These methods show different sensitivity to some of these sources of error in live cell experiments. pH determination based on Raman frequency shift appears to give a more reliable pH determination, though in high signal-to-noise environments, intensity ratios may provide better sensitivity to small changes in pH for cellular imaging.

© 2020 The Author(s)

PDF Article

References

You do not have subscription access to this journal. Citation lists with outbound citation links are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription

Supplementary Material (1)

NameDescription
» Supplement 1       sj-pdf-1-asp-10.1177_0003702820950768 - Supplemental material for Comparison of 4-Mercaptobenzoic Acid Surface-Enhanced Raman Spectroscopy-Based Methods for pH Determination in Cells

Cited By

You do not have subscription access to this journal. Cited by links are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription