Bioanalysis Using Surface-Enhanced Raman Spectroscopy: From the Perspective of Single Molecules

Area of Science:

• Analytical Chemistry
• Spectroscopy
• Biophysics

Background:

• Single-molecule surface-enhanced Raman spectroscopy (SM-SERS) provides unparalleled sensitivity and specificity for probing biomolecular physicochemical properties.
• SM-SERS typically involves complex methodologies, rationally designed substrates, and specialized single-molecule techniques, presenting significant technical hurdles.

Purpose of the Study:

• To critically assess the intrinsic limitations of SM-SERS, moving beyond a comprehensive review.
• To provide a conceptual framework for advancing the fundamental understanding and practical application of SM-SERS.

Main Methods:

• Selective overview of representative SM-SERS methods and features.
• Detailed discussion of overlooked limitations in SM-SERS.

Main Results:

• Identified persistent issues of reproducibility and reliability in SM-SERS.
• Highlighted signal intensity fluctuations (SIFs) and sample degradation as key challenges.
• Addressed the emerging difficulty of big data processing and analysis in SM-SERS.

Conclusions:

• SM-SERS, despite its potential, requires addressing inherent limitations for reliable application.
• Overcoming challenges in signal stability, sample integrity, and data analysis is crucial for advancing SM-SERS.
• A conceptual framework is proposed to guide future research and enhance the practical utility of SM-SERS.