Analysis of subcellular surface structure, function and dynamics.
Analytics of single biological cells allows quantitative investigation from a structural, functional and dynamical point of view and opens novel possibilities to an unamplified subcellular analysis. In this article, we report on three different experimental methods and their applications to single cellular systems with a subcellular sensitivity down to the single molecule level. First, the subcellular surface structure of living bacteria (Corynebacterium glutamicum) was investigated with atomic force microscopy (AFM) at the resolution of individual surface layer (S-layer) proteins; discrimination of bacterial strains that lack the expression of hexagonally packed surface layer proteins was possible. Second, quantitative measurement of individual recognition events of membrane-bound receptors on living B-cells was achieved in single cell manipulation and probing experiments with optical tweezers (OT) force spectroscopy. And third, intracellular dynamics of translocating photoactivatable GFP in plant protoplasts (Nicotiana tabacum BY-2) was quantitatively monitored by two-photon laser scanning microscopy (2PLSM).
Anselmetti
D
D
Experimental Biophysics and Applied Nanoscience, Faculty of Physics, Bielefeld University, Universitätsstrasse 25, 33615, Bielefeld, Germany. dario.anselmetti@physik.uni-bielefeld.de
Hansmeier
N
N
Kalinowski
J
J
Martini
J
J
Merkle
T
T
Palmisano
R
R
Ros
R
R
Schmied
K
K
Sischka
A
A
Toensing
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D001402:B-Lymphocytes / Q000737:chemistry / Q000166:cytology; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D002467:Cell Nucleus / Q000378:metabolism; D002645:Chickens; D048230:Corynebacterium glutamicum / Q000166:cytology / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D047569:Laser Scanning Cytometry / Q000379:methods*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D018625:Microscopy, Atomic Force / Q000379:methods*; D008863:Microspheres; D052898:Optical Tweezers; D021381:Protein Transport; D011523:Protoplasts / Q000166:cytology / Q000378:metabolism; D011947:Receptors, Antigen, B-Cell / Q000737:chemistry / Q000276:immunology / Q000502:physiology; D014026:Nicotiana / Q000166:cytology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D008562:Membrane Glycoproteins; D011947:Receptors, Antigen, B-Cell; C055764:S-layer proteins; D049452:Green Fluorescent Proteins