Risultati scientifici

Three-Dimensional Tracking of Quantum Dot-Conjugated Molecules in Living Cells

Anno: 2018

Autori: Gardini L., Calamai M., Hatakeyama H., Kanzaki M., Capitanio M., Pavone FS.

Affiliazione autori: [Gardini, Lucia; Calamai, Martino; Capitanio, Marco; Pavone, Francesco Saverio] LENS European Lab Nonlinear Spect, Sesto Fiorentino, Italy.
[Gardini, Lucia; Calamai, Martino; Pavone, Francesco Saverio] INO Natl Inst Opt, Sesto Fiorentino, Italy.
[Hatakeyama, Hiroyasu] Tohoku Univ, Frontier Res Inst Interdisciplinary Sci, Sendai, Miyagi, Japan.
[Kanzaki, Makoto] Tohoku Univ, Grad Sch Biomed Engn, Sendai, Miyagi, Japan.
[Capitanio, Marco; Pavone, Francesco Saverio] Univ Florence, Dept Phys & Astron, Sesto Fiorentino, Italy.

Abstract: Here, we describe protocols for three-dimensional tracking of single quantum dot-conjugated molecules with nanometer accuracy in living cells using conventional fluorescence microscopy. The technique exploits out-of-focus images of single emitters combined with an automated pattern-recognition open-source software that fits the images with proper model functions to extract the emitter coordinates. We describe protocols for targeting quantum dots to both membrane components and cytosolic proteins.


Parole chiavi: Cytosolic proteins; Lipids; Living cells; Out-of-focus imaging; Plasma membrane proteins; Quantum dots; Single-molecule localization; Three-dimensional tracking; Ganglioside GM1; Cell labeling; Human; Membrane component

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