Super-resolution.org.au
The Bell Single Molecule and Fluorescence Lab
Correlating Fluorescence:
Live Cell/Fixed Cell Super-Resolution, Atomic Force Microscopy, and Electron Microscopy
Atomic Force Microscopy (AFM, image above) and Scanning Electron Microscopy (SEM) gives a more holistic image, showing the entire environment of the target location which pairs well with dSTORM’s specific structure labelling. When these techniques are combined they can reveal the target structure along with the environment it’s in. Using the Soft Materials and Colloids (SMaC) Lab’s AFM at Monash University the Bell group is currently working on imaging internal cellular structures on both AFM and dSTORM, opening up a wide range of investigations of various cellular perturbation states, including drug induced microtubule polymerisation/depolymerisation.
​
As AFM gives a topographical map of the surface of the sample, in order to view the internal structure of cells the cell membrane needs to be removed first. This invasive sample preparation can easily destroy the internal structure and so live-cell imaging through SOFI can be performed pre-fixation to determine the extent of damage from sample preparation.
SEM could be used instead of AFM however the vacuum conditions require the cell be sealed completely. This can be done using a monolayer of graphene to cover the sample while also creating a conductive layer to shield the cell from the electron beam while keeping the structure intact.
