Continuous throughput and long-term observation of single-molecule FRET without immobilization.
We present an automated microfluidic platform that performs multisecond observation of single molecules with millisecond time resolution while bypassing the need for immobilization procedures. With this system, we confine biomolecules to a thin excitation field by reversibly collapsing microchannels to nanochannels. We demonstrate the power of our method by studying a variety of complex nucleic acid and protein systems, including DNA Holliday junctions, nucleosomes and human transglutaminase 2.
1] Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany. [2].
Swati
S
Tyagi
1] Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany. [2] [3].
Virginia
V
VanDelinder
Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Niccolò
N
Banterle
Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Gustavo
G
Fuertes
Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Sigrid
S
Milles
Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Morgane
M
Agez
Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Edward A
EA
Lemke
D001331:Automation; D031541:Fluorescence Resonance Energy Transfer; D019204:GTP-Binding Proteins / Q000235:genetics; D006801:Humans; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D008958:Models, Molecular; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases / Q000235:genetics
D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins