ABSTRACT
Biomolecule imaging within the cell enables us to study the molecular mechanism and cellular response. Single-molecule imaging is a technique that investigates the properties of the individual molecular responses of the biological system. Conventional imaging techniques like light, electron, and fluorescence microscopy are used in cell biology to observe the biological system. These imaging techniques require exogenous contrast agents like fluorophores and nanoprobes to improve the imaging parameters. The techniques combining fluorescence light microscopy and electron microscopy for imaging the structures of the biomolecules enable good single-molecule imaging. The use of fluorescent tags for imaging provides high labeling specificity to detect and analyze individual single molecules. This chapter begins with the principle and needs for the single-molecule imaging technique. Focuses on the design of single-molecule detection techniques aided by imaging modalities, fluorescent probes, and labeling methods. Also, this chapter emphasizes the quantitative aspects of the imaging modality, image formation, processing, and different analytical estimation techniques for visualization. The single-molecule imaging technique accomplished various development by different biophysics research groups. Overall, the different imaging techniques enable single-molecule detection with their instrumentation and application using biomolecules in the field of biomedical engineering.
