TY - CHAP
T1 - Three-dimensional tracking of quantum dot-conjugated molecules in living cells
AU - Gardini, Lucia
AU - Calamai, Martino
AU - Hatakeyama, Hiroyasu
AU - Kanzaki, Makoto
AU - Capitanio, Marco
AU - Pavone, Francesco Saverio
PY - 2018/1/1
Y1 - 2018/1/1
N2 - 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.
AB - 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.
KW - Cytosolic proteins
KW - Lipids
KW - Living cells
KW - Out-of-focus imaging
KW - Plasma membrane proteins
KW - Quantum dots
KW - Single-molecule localization
KW - Three-dimensional tracking
UR - http://www.scopus.com/inward/record.url?scp=85049332705&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85049332705&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-8591-3_26
DO - 10.1007/978-1-4939-8591-3_26
M3 - Chapter
C2 - 29956248
AN - SCOPUS:85049332705
T3 - Methods in Molecular Biology
SP - 425
EP - 448
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -