A Non-Destructive, Tuneable Method to Isolate Live Cells for High-Speed AFM Analysis

Microorganisms. 2021 Mar 25;9(4):680. doi: 10.3390/microorganisms9040680.

Abstract

Suitable immobilisation of microorganisms and single cells is key for high-resolution topographical imaging and study of mechanical properties with atomic force microscopy (AFM) under physiologically relevant conditions. Sample preparation techniques must be able to withstand the forces exerted by the Z range-limited cantilever tip, and not negatively affect the sample surface for data acquisition. Here, we describe an inherently flexible methodology, utilising the high-resolution three-dimensional based printing technique of multiphoton polymerisation to rapidly generate bespoke arrays for cellular AFM analysis. As an example, we present data collected from live Emiliania huxleyi cells, unicellular microalgae, imaged by contact mode High-Speed Atomic Force Microscopy (HS-AFM), including one cell that was imaged continuously for over 90 min.

Keywords: 3D printing; atomic force microscopy; high-speed; immobilization; microalgae; microbe; multiphoton polymerization.