Affinity imaging of red blood cells using an atomic force microscope.
We used an atomic force microscope (AFM) to produce an image of a mixed layer of group A and O red blood cells with a contrast based only on the measured strength of a specific receptor-ligand pair. The image was obtained by measuring and plotting for each image pixel the adhesion force between the mixed RBC layer and the AFM tip functionalized with Helix pomatia lectin. The high specificity of that lectin for the N -acetylgalactosamine-terminated glycolipids present in the membrane of group A RBCs enabled us to discriminate between the two cell populations and to produce an image based on affinity contrast. The rupture force of the adhesion events leading to the image formation were quantitatively analyzed and compared to rupture forces measured with the same AFM tip on N-acetylgalactosamine tethered to agarose beads. The mean rupture force was found to be 65 pN when measured on the group A RBCs and 35 pN on the agarose beads. These results show that the adhesion, mediated by only a few receptor-ligand pairs, produces sufficient contrast for the affinity image formation.
Grandbois
M
M
Department of Physics and Astronomy, University of Missouri-Columbia, Columbia, Missouri 65211, USA. grandboism@missouri.edu
Dettmann
W
W
Benoit
M
M
Gaub
H E
HE
D000017:ABO Blood-Group System / Q000378:metabolism; D000116:Acetylgalactosamine / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D004912:Erythrocytes / Q000166:cytology* / Q000502:physiology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D037102:Lectins / Q000378:metabolism; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D012685:Sepharose / Q000378:metabolism; D013379:Substrate Specificity
D000017:ABO Blood-Group System; C038095:Helix lectin; D037102:Lectins; D012685:Sepharose; D000116:Acetylgalactosamine