Selected Single-Molecule Experiments with Relevance to Living Cells
(bold entries in living cells)
Modified from: Lord, S. J.; Lee, H. D.; Moerner, W. E. Single-Molecule Spectroscopy and Imaging of Biomolecules in Living Cells. Anal. Chem. 2010, 82(6), 2192-2203. (HTML, PDF)
|
year |
milestone |
primary group leaders |
references |
|
1961 |
catalytic activity from one enzyme measured (by observing the buildup of fluorescence from a fluorogenic substrate) |
Rotman |
5 |
| 1978 | optical detection of a single polymer molecule (labeled with many fluorophores) | Hirschfeld | 125 |
|
1981 |
single strands of DNA visualized (labeled with many fluorophores) |
Yanagida, Chu |
6, 7 |
|
1982 |
diffusion of single membrane-bound receptor molecules measured (labeled with many fluorophores) |
W. Webb |
8 |
|
1986 |
single actin filaments (labeled with many fluorophores) sliding on myosin coverslips |
Spudich |
127 |
| 1988 | tracking movements with nm precision | Sheetz | 126 |
|
1989, 1990 |
first optical detection of single chromophore in condensed matter (cryogenic) |
Moerner, Orrit |
13−15 |
|
action from single molecular motors recorded by observing motion of filaments (e.g., sliding assays) |
Vale, Yanagida, Spudich |
117 |
|
|
1990 |
room-temperature detection of single molecules from burst analysis in solution |
Keller, Rigler, Zare |
16−18 |
|
1991 |
first imaging of a single molecule (cryogenic) |
Moerner |
15, 99 |
|
1992 |
elastic measurements of single strands of DNA using magnetic tweezers |
Bustamante, Croquette, Bensimon |
1, 3 |
|
1993 |
room-temperature SMS using near-field tips |
Betzig |
20 |
|
room-temperature SMS in microdroplets |
Ramsey |
19 |
|
|
force studies of single macromolecules using optical tweezers or cantilevers |
Block, Gaub |
2, 4 |
|
|
1995 |
single motor molecules imaged |
Yanagida, Kinosita, Vale |
27−29 |
|
1996 |
3D nanoscale tracking of single emitters, using TIRF evanescent field |
Moerner |
21 |
|
first single-pair FRET measurements |
Weiss, Ha |
25 |
|
|
diffusion of single emitters recorded in a phospholipid membrane |
Schmidt, Schindler |
26 |
|
|
1997 |
first SMS of fluorescent proteins |
Moerner, Tsien, Vale |
24, 118 |
|
first room-temperature SMS example of controlled photoswitching, a fluorescent protein |
Moerner, Tsien |
24 |
|
|
1998 |
3D orientations of single molecules measured from dipole emission pattern (room temperature) |
Moerner |
119 |
|
3D super-resolution by spectral selection of single molecules (cryogenic) |
Betzig, van Oijen, Schmidt, Moerner |
90, 99−101 |
|
| single-molecule enzymatic turnovers of flavoenzyme | Xie, Lu | 124 | |
|
2000 |
spFRET measured in living cells |
Yanagida, Kusumi, S. Webb |
96−98 |
|
3D tracking of single fluorophores in living cells |
Schuetz, Schindler |
78 |
|
|
transmembrane ion channels tracked |
Schmidt, Schuetz, Schindler |
77, 78 |
|
|
2001 |
nucleotide-dependent orientations of single kinesin motors measured |
Moerner, Goldstein |
30 |
|
binding kinetics to chemotactic receptors in the membrane observed |
Yanagida |
79 |
|
|
infection pathway of singly labeled viruses observed |
Brauchle |
91 |
|
|
2002 |
SMS in membranes |
Moerner, McConnell, Kusumi, Dahan, Triller |
67−69, 71−74, 76 |
|
2003 |
single motor proteins tracked with high precision and super-resolution |
Selvin, Spudich |
120, 121 |
|
2004 |
protein localization and movement in living bacteria cells using FP labels |
Moerner, Shapiro, McAdams |
35, 85−87 |
|
molecules tracked through nuclear pore complexes |
Musser |
82 |
|
|
2005 |
nerve growth factor tracked in living neurons |
Tani, Yanagida, Chu, Mobley, Cui |
89, 122 |
|
2006 |
high-precision tracing of filaments in living bacteria |
Moerner, Shapiro |
35 |
|
super-resolution SMS techniques PALM, STORM, and FPALM introduced |
Betzig, H. Hess, Lippincott-Schwartz, Zhuang, S. Hess |
102−104 |
|
|
single-molecule gene expression events studied in living bacteria |
Xie |
36, 94, 95 |
|
|
singly labeled molecular motors observed in living cells |
Meyhoefer, Dahan, Cappello, Spudich |
92, 93, 123 |
|
|
2007 |
super-resolution SMS images of living cells |
Betzig, H. Hess, Lippincott-Schwartz, S. Hess, Moerner |
54, 87, 107−109 |
|
SMS used to count the number of subunits in membrane-bound proteins |
Isacoff, Verkman |
80, 81 |
|
|
2008 |
interaction of cell-penetrating peptides with membrane observed |
Moerner, Wender |
75 |
|
single monomers of the cytoskeletal protein photoactivated and tracked |
Yu |
88 |
|
|
2009 |
SMS in living vertebrates |
Schmidt, Schaaf |
110 |
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