Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy

O H Willemsen, M M Snel, K O van der Werf, B G de Grooth, J Greve, P Hinterdorfer, H J Gruber, H Schindler, Y van Kooyk, C G Figdor

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Specific molecular recognition events, detected by atomic force microscopy (AFM), so far lack the detailed topographical information that is usually observed in AFM. We have modified our AFM such that, in combination with a recently developed method to measure antibody-antigen recognition on the single molecular level (Hinterdorfer, P., W. Baumgartner, H. J. Gruber, K. Schilcher, and H. Schindler, Proc. Natl. Acad. Sci. USA 93:3477-3481 (1996)), it allows imaging of a submonolayer of intercellular adhesion molecule-1 (ICAM-1) in adhesion mode. We demonstrate that for the first time the resolution of the topographical image in adhesion mode is only limited by tip convolution and thus comparable to tapping mode images. This is demonstrated by imaging of individual ICAM-1 antigens in both the tapping mode and the adhesion mode. The contrast in the adhesion image that was measured simultaneously with the topography is caused by recognition between individual antibody-antigen pairs. By comparing the high-resolution height image with the adhesion image, it is possible to show that specific molecular recognition is highly correlated with topography. The stability of the improved microscope enabled imaging with forces as low as 100 pN and ultrafast scan speed of 22 force curves per second. The analysis of force curves showed that reproducible unbinding events on subsequent scan lines could be measured.

Original languageEnglish
Pages (from-to)2220-8
Number of pages9
JournalBiophysical Journal
Volume75
Issue number5
DOIs
Publication statusPublished - Nov 1998

Cite this

Willemsen, O. H., Snel, M. M., van der Werf, K. O., de Grooth, B. G., Greve, J., Hinterdorfer, P., ... Figdor, C. G. (1998). Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy. Biophysical Journal, 75(5), 2220-8. https://doi.org/10.1016/S0006-3495(98)77666-0
Willemsen, O H ; Snel, M M ; van der Werf, K O ; de Grooth, B G ; Greve, J ; Hinterdorfer, P ; Gruber, H J ; Schindler, H ; van Kooyk, Y ; Figdor, C G. / Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy. In: Biophysical Journal. 1998 ; Vol. 75, No. 5. pp. 2220-8.
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title = "Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy",
abstract = "Specific molecular recognition events, detected by atomic force microscopy (AFM), so far lack the detailed topographical information that is usually observed in AFM. We have modified our AFM such that, in combination with a recently developed method to measure antibody-antigen recognition on the single molecular level (Hinterdorfer, P., W. Baumgartner, H. J. Gruber, K. Schilcher, and H. Schindler, Proc. Natl. Acad. Sci. USA 93:3477-3481 (1996)), it allows imaging of a submonolayer of intercellular adhesion molecule-1 (ICAM-1) in adhesion mode. We demonstrate that for the first time the resolution of the topographical image in adhesion mode is only limited by tip convolution and thus comparable to tapping mode images. This is demonstrated by imaging of individual ICAM-1 antigens in both the tapping mode and the adhesion mode. The contrast in the adhesion image that was measured simultaneously with the topography is caused by recognition between individual antibody-antigen pairs. By comparing the high-resolution height image with the adhesion image, it is possible to show that specific molecular recognition is highly correlated with topography. The stability of the improved microscope enabled imaging with forces as low as 100 pN and ultrafast scan speed of 22 force curves per second. The analysis of force curves showed that reproducible unbinding events on subsequent scan lines could be measured.",
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Willemsen, OH, Snel, MM, van der Werf, KO, de Grooth, BG, Greve, J, Hinterdorfer, P, Gruber, HJ, Schindler, H, van Kooyk, Y & Figdor, CG 1998, 'Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy' Biophysical Journal, vol. 75, no. 5, pp. 2220-8. https://doi.org/10.1016/S0006-3495(98)77666-0

Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy. / Willemsen, O H; Snel, M M; van der Werf, K O; de Grooth, B G; Greve, J; Hinterdorfer, P; Gruber, H J; Schindler, H; van Kooyk, Y; Figdor, C G.

In: Biophysical Journal, Vol. 75, No. 5, 11.1998, p. 2220-8.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy

AU - Willemsen, O H

AU - Snel, M M

AU - van der Werf, K O

AU - de Grooth, B G

AU - Greve, J

AU - Hinterdorfer, P

AU - Gruber, H J

AU - Schindler, H

AU - van Kooyk, Y

AU - Figdor, C G

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N2 - Specific molecular recognition events, detected by atomic force microscopy (AFM), so far lack the detailed topographical information that is usually observed in AFM. We have modified our AFM such that, in combination with a recently developed method to measure antibody-antigen recognition on the single molecular level (Hinterdorfer, P., W. Baumgartner, H. J. Gruber, K. Schilcher, and H. Schindler, Proc. Natl. Acad. Sci. USA 93:3477-3481 (1996)), it allows imaging of a submonolayer of intercellular adhesion molecule-1 (ICAM-1) in adhesion mode. We demonstrate that for the first time the resolution of the topographical image in adhesion mode is only limited by tip convolution and thus comparable to tapping mode images. This is demonstrated by imaging of individual ICAM-1 antigens in both the tapping mode and the adhesion mode. The contrast in the adhesion image that was measured simultaneously with the topography is caused by recognition between individual antibody-antigen pairs. By comparing the high-resolution height image with the adhesion image, it is possible to show that specific molecular recognition is highly correlated with topography. The stability of the improved microscope enabled imaging with forces as low as 100 pN and ultrafast scan speed of 22 force curves per second. The analysis of force curves showed that reproducible unbinding events on subsequent scan lines could be measured.

AB - Specific molecular recognition events, detected by atomic force microscopy (AFM), so far lack the detailed topographical information that is usually observed in AFM. We have modified our AFM such that, in combination with a recently developed method to measure antibody-antigen recognition on the single molecular level (Hinterdorfer, P., W. Baumgartner, H. J. Gruber, K. Schilcher, and H. Schindler, Proc. Natl. Acad. Sci. USA 93:3477-3481 (1996)), it allows imaging of a submonolayer of intercellular adhesion molecule-1 (ICAM-1) in adhesion mode. We demonstrate that for the first time the resolution of the topographical image in adhesion mode is only limited by tip convolution and thus comparable to tapping mode images. This is demonstrated by imaging of individual ICAM-1 antigens in both the tapping mode and the adhesion mode. The contrast in the adhesion image that was measured simultaneously with the topography is caused by recognition between individual antibody-antigen pairs. By comparing the high-resolution height image with the adhesion image, it is possible to show that specific molecular recognition is highly correlated with topography. The stability of the improved microscope enabled imaging with forces as low as 100 pN and ultrafast scan speed of 22 force curves per second. The analysis of force curves showed that reproducible unbinding events on subsequent scan lines could be measured.

KW - Aluminum Silicates/chemistry

KW - Animals

KW - Antibodies, Monoclonal/ultrastructure

KW - Antigen-Antibody Complex/ultrastructure

KW - Binding Sites/physiology

KW - CHO Cells

KW - Cell Adhesion/physiology

KW - Cell Adhesion Molecules/ultrastructure

KW - Cricetinae

KW - Humans

KW - Image Processing, Computer-Assisted

KW - Intercellular Adhesion Molecule-1/ultrastructure

KW - Microscopy, Atomic Force/methods

KW - Peptide Fragments/ultrastructure

KW - Protein Binding

U2 - 10.1016/S0006-3495(98)77666-0

DO - 10.1016/S0006-3495(98)77666-0

M3 - Article

VL - 75

SP - 2220

EP - 2228

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 5

ER -

Willemsen OH, Snel MM, van der Werf KO, de Grooth BG, Greve J, Hinterdorfer P et al. Simultaneous height and adhesion imaging of antibody-antigen interactions by atomic force microscopy. Biophysical Journal. 1998 Nov;75(5):2220-8. https://doi.org/10.1016/S0006-3495(98)77666-0