A sticky tale: sample preparation technique determines cell surface receptor mobility and adhesion

Müller, Thomas; Marshall, Jennifer; Dehghani, Hamid; Bowen, James and Toellner, Kai-Michael (2014). A sticky tale: sample preparation technique determines cell surface receptor mobility and adhesion. In: 6th AFMBioMed Conference, 13-17 Dec 2014, San Diego, USA.

URL: http://www.researchgate.net/publication/269810552_...

Abstract

Cell surface proteins, for example specific antigen receptors on lymphocytes, have become common targets for interrogation via atomic force microscopy (AFM). While the majority of studies use live cells in order to mimic in vivo conditions, this is not always feasible. For successful AFM adhesion measurements, cells that exhibit poor natural adhesion to a counter-surface need to be immobilised, or have their mobility restricted. To be able to interrogate structures within tissue sections – for example germinal centres in spleen sections – fixation is required for preserving the structure of the tissue for the long duration of the interrogation, as well as preserving any applied immunohistochemical staining necessary for identification of regions of interest. The work presented here investigates the effects that the commonly used fixatives acetone and paraformaldehyde (PFA) have on biological samples, and their subsequent influences on interactions between AFM tip and cell surface proteins in adhesion measurements.

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