Low Stress Ion Conductance Microscopy of Sub-Cellular Stiffness.
Directly examining subcellular mechanics whilst avoiding excessive strain of a live cell requires the precise control of light stress on very small areas, which is fundamentally difficult. Here we use a glass nanopipet out of contact with the plasma membrane to both exert the stress on the cell and also accurately monitor cellular compression. This allows the mapping of cell stiffness at a lateral resolution finer than 100 nm. We calculate the stress a nanopipet exerts on a cell as the sum of the intrinsic pressure between the tip face and the plasma membrane plus its direct pressure on any glycocalyx, both evaluated from the gap size in terms of the ion current decrease. A survey of cell types confirms that an intracellular pressure of approximately 120 Pa begins to detach the plasma membrane from the cytoskeleton and reveals that the first 0.66 ± 0.09 μm of compression of a neuron cell body is much softer than previous methods have been able to detect.
University Chemical Laboratories, Lensfield Road, Cambridge, CB2 1EW, UK. rwc25@cam.ac.uk dk10012@cam.ac.uk.
Richard W
RW
Clarke
Pavel
P
Novak
Alexander
A
Zhukov
Eleanor J
EJ
Tyler
Marife
M
Cano-Jaimez
Anna
A
Drews
Owen
O
Richards
Kirill
K
Volynski
Cleo
C
Bishop
David
D
Klenerman
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology*; D002478:Cells, Cultured; D003593:Cytoplasm; D003599:Cytoskeleton; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007477:Ions; D008853:Microscopy / Q000379:methods*; D009474:Neurons / Q000166:cytology; D051381:Rats
D007477:Ions