Direct manipulation of intracellular stress fibres using a hook-shaped AFM probe.
Atomic force microscopy (AFM) is a highly successful technique for imaging nanometre-sized samples and measuring pico- to nano-newton forces acting between atoms and molecules. When it comes to the manipulation of larger samples with forces of tens and hundreds of nano-newtons, however, the present chemistry-based modification protocols for functionalizing AFM cantilevers to achieve the formation of covalent/non-covalent linkages between the AFM probe and the sample surface do not produce strong enough bonds. For the purpose of measuring the fracture strength and other mechanical properties of stress fibres (SFs) in living as well as semi-intact fibroblast cells, we fabricated an AFM probe with a hooking function by focused ion beam technology and used the AFM probe hook to capture, pull and eventually sever a chosen SF labelled with green or red fluorescent protein.
Machida
Shinichi
S
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama, Kanagawa, Japan. smachida@bio.titech.ac.jp
Watanabe-Nakayama
Takahiro
T
Harada
Ichiro
I
Afrin
Rehana
R
Nakayama
Tomonobu
T
Ikai
Atsushi
A
D000818:Animals; D002478:Cells, Cultured; D005347:Fibroblasts / Q000648:ultrastructure*; D049452:Green Fluorescent Proteins / Q000032:analysis; D008164:Luminescent Proteins / Q000032:analysis; D055595:Mechanical Phenomena; D018625:Microscopy, Atomic Force / Q000295:instrumentation* / Q000379:methods; D051381:Rats; D022502:Stress Fibers / Q000737:chemistry / Q000648:ultrastructure*; D000097573:Red Fluorescent Protein
D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein