Second Harmonic Atomic Force Microscopy Imaging of Live and Fixed Mammalian Cells

Alexander Dulebo, Johannes Preiner, Ferry Kienberger, Gerald Kada, Christian Rankl, Lilia Chtcheglova, Constanze Lamprecht, David Kaftan, Peter Hinterdorfer

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Higherharmoniccontributionsinthemovementofanoscillatingatomicforcemicroscopy(AFM) cantileveraregeneratedbynonlineartip–sampleinteractions,yieldingadditionalinformationonstructure andphysicalpropertiessuchassamplestiffness.Higherharmonicamplitudesarestronglyenhancedin liquidcomparedtotheoperationinair,andwerepreviouslyreportedtoresultinbetterstructural resolutioninhighlyorganizedlatticesofproteinsinbacterialS-layersandviralcapsids[J.Preiner,J.Tang, V. Pastushenko,P.Hinterdorfer,Phys.Rev.Lett.99(2007)046102].Wecomparedfirstandsecond harmonicsAFMimagingofliveandfixedhumanlungepithelialcells,andmicrovascularendothelialcells frommousemyocardium(MyEnd).Phase–distancecyclesrevealedthatthesecondharmonicphaseis8 timesmoresensitivethanthefirstharmonicphasewithrespecttovariationsinthedistancebetween cantileverandsamplesurface.Frequencyspectrawereacquiredatdifferentpositionsonlivingandfixed cellswithsecondharmonicamplitudevaluescorrelatingwiththesamplestiffness.Weconcludethat variationsinsamplestiffnessandcorrespondingchangesinthecantilever–sampledistance,lattereffect causedbythefinitefeedbackresponse,resultinsecondharmonicimageswithimprovedcontrastand informationthatisnotattainableinthefundamentalfrequencyofanoscillatingcantilever.
Original languageEnglish
Pages (from-to)1056-1060
Number of pages5
JournalUltramicroscopy
Volume109
Issue number8
DOIs
Publication statusPublished - Jul 2009

Keywords

  • Atomic force microscopy
  • Cells
  • Higher harmonic
  • Live cell imaging

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