TY - JOUR
T1 - Second Harmonic Atomic Force Microscopy Imaging of Live and Fixed Mammalian Cells
AU - Dulebo, Alexander
AU - Preiner, Johannes
AU - Kienberger, Ferry
AU - Kada, Gerald
AU - Rankl, Christian
AU - Chtcheglova, Lilia
AU - Lamprecht, Constanze
AU - Kaftan, David
AU - Hinterdorfer, Peter
N1 - Funding Information:
The present study was supported by grants from the Swedish Cancer Society, the Medical Faculty, UmeI University, and Lions Research Foundation, Umeb, Sweden.
PY - 2009/7
Y1 - 2009/7
N2 - 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.
AB - 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.
KW - Atomic force microscopy
KW - Cells
KW - Higher harmonic
KW - Live cell imaging
UR - http://www.scopus.com/inward/record.url?scp=67449146904&partnerID=8YFLogxK
U2 - 10.1016/j.ultramic.2009.03.020
DO - 10.1016/j.ultramic.2009.03.020
M3 - Article
SN - 1879-2723
VL - 109
SP - 1056
EP - 1060
JO - Ultramicroscopy
JF - Ultramicroscopy
IS - 8
ER -