AFM-Based Correlative Microscopy Illuminates Human Pathogens

Abstract

Microbes have an arsenal of virulence factors that contribute to their pathogenicity. A number of challenges remain to fully understand disease transmission, fitness landscape, antimicrobial resistance and host heterogeneity. A variety of tools have been used to address diverse aspects of pathogenicity, from molecular host-pathogen interactions to the mechanisms of disease acquisition and transmission. Current gaps in our knowledge include a more direct understanding of host-pathogen interactions, including signaling at interfaces, and direct phenotypic confirmation of pathogenicity. Correlative microscopy has been gaining traction to address the many challenges currently faced in biomedicine, in particular the combination of optical and atomic force microscopy (AFM). AFM, generates high-resolution surface topographical images, and quantifies mechanical properties at the pN scale under physiologically relevant conditions. When combined with optical microscopy, AFM probes pathogen surfaces and their physical and molecular interaction with host cells, while the various modes of optical microscopy view internal cellular responses of the pathogen and host. Here we review the most recent advances in our understanding of pathogens, recent applications of AFM to the field, how correlative AFM-optical microspectroscopy and microscopy have been used to illuminate pathogenicity and how these methods can reach their full potential for studying host-pathogen interactions.

The Roles of Optogenetics and Technology in Neurobiology: A Review

“…Researchers are also developing more precise ( Adesnik and Abdeladim, 2021 ) and less invasive optogenetic devices, such as SOUL ( Gong et al, 2020 ). At the same time, optogenetics has strong compatibility, it can be used to study a variety of diseases, such as diabetes ( Li et al, 2021 ), inflammation ( Baumschlager and Khammash, 2021 ; Bhat et al, 2021 ; Dos Santos et al, 2021 ; Jamaluddin et al, 2021 ; Michoud et al, 2021 ; Santos et al, 2021 ; Senok et al, 2021 ), tumors ( Kim et al, 2017 ; Adampourezare et al, 2021 ; Esmaeili et al, 2022 ), depression ( Hare et al, 2019 ), epilepsy ( Zhang and Wang, 2021 ) and so on. And in 2020, 45 laboratories around the world integrated all optogenetics resources and create an optogenetics experimental database ( Tremblay et al, 2020 ).…”

Applicability of atomic force microscopy to determine cancer-related changes in cells

“…The use of AFM in more complex systems like spheroids [76] or patient-derived tissues [26,77] reveals the importance of mechanical properties not only of a single-cell but also ECM, in which cells are embedded. Thus, valuable development directions guide towards techniques combined directly or in parallel, in which the similar (or the same) sample will be studied using not only AFM but also other techniques, such as scanning electron or confocal or super-resolution fluorescent microscopes [78][79][80] or Raman or FTIR spectroscopy [81,82].…”

Extending applications of AFM to fluidic AFM in single living cell studies

“…Invented by Binnig et al (Binnig et al, 1986), atomic force microscopy (AFM) has emerged as an indispensable technique for the study of the biomechanical features of the cell surface in real time (Bhat et al, 2021). AFM measures the force dynamics between a probe tip and a sample at the nanoscale level.…”