Scanning ion conductance microscopy of live keratinocytes

V. Hegde, A. Mason, T. Saliev, F. J D Smith, W. H I McLean, P. A. Campbell

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Scanning ion conductance microscopy (SICM) is perhaps the least well known technique from the scanning probe microscopy (SPM) family of instruments. As with its more familiar counterpart, atomic force microscopy (AFM), the technique provides high-resolution topographic imaging, with the caveat that target structures must be immersed in a conducting solution so that a controllable ion current may be utilised as the basis for feedback. In operation, this non-contact characteristic of SICM makes it ideal for the study of delicate structures, such as live cells. Moreover, the intrinsic architecture of the instrument, incorporating as it does, a scanned micropipette, lends itself to combination approaches with complementary techniques such as patch-clamp electrophysiology: SICM therefore boasts the capability for both structural and functional imaging. For the present observations, an ICnano S system (Ionscope Ltd., Melbourn, UK) operating in 'hopping mode' was used, with the objective of assessing the instrument's utility for imaging live keratinocytes under physiological buffers. In scans employing cultured HaCaT cells (spontaneously immortalised, human keratinocytes), we compared the qualitative differences of live cells imaged with SICM and AFM, and also with their respective counterparts after chemical fixation in 4% paraformaldehyde. Characteristic surface microvilli were particularly prominent in live cell imaging by SICM. Moreover, time lapse SICM imaging on live cells revealed that changes in the pattern of microvilli could be tracked over time. By comparison, AFM imaging on live cells, even at very low contact forces (

Original languageEnglish
Article number012023
JournalJournal of Physics: Conference Series
Volume371
DOIs
Publication statusPublished - 2012
Externally publishedYes

Fingerprint

microscopy
scanning
ions
cells
atomic force microscopy
electrophysiology
clamps
cultured cells
ion currents
buffers
conduction
probes
high resolution

ASJC Scopus subject areas

  • Physics and Astronomy(all)

Cite this

Hegde, V., Mason, A., Saliev, T., Smith, F. J. D., McLean, W. H. I., & Campbell, P. A. (2012). Scanning ion conductance microscopy of live keratinocytes. Journal of Physics: Conference Series, 371, [012023]. https://doi.org/10.1088/1742-6596/371/1/012023

Scanning ion conductance microscopy of live keratinocytes. / Hegde, V.; Mason, A.; Saliev, T.; Smith, F. J D; McLean, W. H I; Campbell, P. A.

In: Journal of Physics: Conference Series, Vol. 371, 012023, 2012.

Research output: Contribution to journalArticle

Hegde, V. ; Mason, A. ; Saliev, T. ; Smith, F. J D ; McLean, W. H I ; Campbell, P. A. / Scanning ion conductance microscopy of live keratinocytes. In: Journal of Physics: Conference Series. 2012 ; Vol. 371.
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