Progress in the Correlative Atomic Force Microscopy and Optical Microscopy

Abstract

Atomic force microscopy (AFM) has evolved from the originally morphological imaging technique to a powerful and multifunctional technique for manipulating and detecting the interactions between molecules at nanometer resolution. However, AFM cannot provide the precise information of synchronized molecular groups and has many shortcomings in the aspects of determining the mechanism of the interactions and the elaborate structure due to the limitations of the technology, itself, such as non-specificity and low imaging speed. To overcome the technical limitations, it is necessary to combine AFM with other complementary techniques, such as fluorescence microscopy. The combination of several complementary techniques in one instrument has increasingly become a vital approach to investigate the details of the interactions among molecules and molecular dynamics. In this review, we reported the principles of AFM and optical microscopy, such as confocal microscopy and single-molecule localization microscopy, and focused on the development and use of correlative AFM and optical microscopy.

Preferential interactions of primary amine-terminated quantum dots with membrane domain boundaries and lipid rafts revealed with nanometer resolution

“…20,24,27 While not attaining the resolution of AFM, fluorescence microscopy provides high detection selectivity enabled by fluorescence tagging. 28 Correlative AFM and fluorescence microscopy can localize NPs in relation to other fluorescently tagged biomolecules with very high spatial resolution and detection selectivity. The correlative approach has been used to characterize phase segregation in SLBs 29,30 and membrane restructuring due to the addition of ceramide, 31,32 as well as to study the impact of 2,4-dichlorophenoxyacetic acid on bacterial, fungal, and human cells 33 and localize labeled fusion proteins within mammalian cells.…”

Visualization of Cell Structure by Atomic Force Microscopy

“…Current research mainly is focused on the understanding of biological processes at the nanoscale, as recognition of molecules and their properties with no labeling and the interaction among them [22][23][24]. This mini review revealed that only few papers deals with inner cell structure in situ.…”

Correlated Multimodal Imaging in Life Sciences: Expanding the Biomedical Horizon

“…Sample preparation procedures for exvivo microscopy differ substantially across the technologies and even within a modality; AFM images alone, for example, can be acquired under various conditions (vacuum, atmosphere, and liquid). Correlative imaging usually requires modality-specific preparation and setup trade-offs, such as between preservation of fluorescence and subcellular architecture for CLEM, between the AFM laser and excitation spectra of the used fluorophores to avoid bleaching for correlative AFM [78], or between preservation of fluorescence and X-ray contrast for correlative CT. Dependent on the used technology, the sample preparation needs to be adapted.…”