Three-dimensional imaging of single molecules solvated in pores of poly(acrylamide) gels.
Individual fluorescent molecules and individual singly labeled proteins were observed in the water-filled pores of poly(acrylamide) gels by far-field microscopy. Brownian motion was markedly reduced by the gel framework, thus enabling extended study of single fluorophores in aqueous environments. A highly axially dependent laser field was used both to excite the fluorophores and to image the molecules in three dimensions. Single molecules were followed as they moved within and through the porous gel structure. In contrast to dry polymeric hosts, these water-based gels may form a useful medium for single-molecule studies of biological systems in vitro.
Dickson
R M
RM
Department of Chemistry and Biochemistry, Mail Code 0340, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0340, USA.
Norris
D J
DJ
Tzeng
Y L
YL
Moerner
W E
WE
D000180:Acrylic Resins / Q000737:chemistry*; D000888:Antibodies, Anti-Idiotypic / Q000032:analysis* / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D005782:Gels; D007074:Immunoglobulin G / Q000276:immunology; D007834:Lasers; D008856:Microscopy, Fluorescence; D010078:Oxazines / Q000032:analysis* / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D012996:Solutions
D000180:Acrylic Resins; D000888:Antibodies, Anti-Idiotypic; D005456:Fluorescent Dyes; D005782:Gels; D007074:Immunoglobulin G; D010078:Oxazines; D011506:Proteins; D012996:Solutions; C016680:polyacrylamide gels; C044808:nile red