Resolving bundled microtubules using anti-tubulin nanobodies
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Resolving bundled microtubules using anti-tubulin nanobodies. / Mikhaylova, Marina; Cloin, Bas M.C.; Finan, Kieran; van den Berg, Robert; Teeuw, Jalmar; Kijanka, Marta M.; Sokolowski, Mikolaj; Katrukha, Eugene A.; Maidorn, Manuel; Opazo, Felipe; Moutel, Sandrine; Vantard, Marilyn; Perez, Frank; van Bergen en Henegouwen, Paul M. P.; Hoogenraad, Casper C.; Ewers, Helge; Kapitein, Lukas C.
in: NAT COMMUN, Jahrgang 6, Nr. 7933, 11.08.2015.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Resolving bundled microtubules using anti-tubulin nanobodies
AU - Mikhaylova, Marina
AU - Cloin, Bas M.C.
AU - Finan, Kieran
AU - van den Berg, Robert
AU - Teeuw, Jalmar
AU - Kijanka, Marta M.
AU - Sokolowski, Mikolaj
AU - Katrukha, Eugene A.
AU - Maidorn, Manuel
AU - Opazo, Felipe
AU - Moutel, Sandrine
AU - Vantard, Marilyn
AU - Perez, Frank
AU - van Bergen en Henegouwen, Paul M. P.
AU - Hoogenraad, Casper C.
AU - Ewers, Helge
AU - Kapitein, Lukas C.
PY - 2015/8/11
Y1 - 2015/8/11
N2 - Microtubules are hollow biopolymers of 25-nm diameter and are key constituents of the cytoskeleton. In neurons, microtubules are organized differently between axons and dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techniques can detect specific structures at an increased resolution, but the narrow spacing between neuronal microtubules poses challenges because most existing labelling strategies increase the effective microtubule diameter by 20-40 nm and will thereby blend neighbouring microtubules into one structure. Here we develop single-chain antibody fragments (nanobodies) against tubulin to achieve super-resolution imaging of microtubules with a decreased apparent diameter. To test the resolving power of these novel probes, we generate microtubule bundles with a known spacing of 50-70 nm and successfully resolve individual microtubules. Individual bundled microtubules can also be resolved in different mammalian cells, including hippocampal neurons, allowing novel insights into fundamental mechanisms of microtubule organization in cell- and neurobiology.
AB - Microtubules are hollow biopolymers of 25-nm diameter and are key constituents of the cytoskeleton. In neurons, microtubules are organized differently between axons and dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techniques can detect specific structures at an increased resolution, but the narrow spacing between neuronal microtubules poses challenges because most existing labelling strategies increase the effective microtubule diameter by 20-40 nm and will thereby blend neighbouring microtubules into one structure. Here we develop single-chain antibody fragments (nanobodies) against tubulin to achieve super-resolution imaging of microtubules with a decreased apparent diameter. To test the resolving power of these novel probes, we generate microtubule bundles with a known spacing of 50-70 nm and successfully resolve individual microtubules. Individual bundled microtubules can also be resolved in different mammalian cells, including hippocampal neurons, allowing novel insights into fundamental mechanisms of microtubule organization in cell- and neurobiology.
U2 - doi: 10.1038/ncomms8933
DO - doi: 10.1038/ncomms8933
M3 - SCORING: Journal article
VL - 6
JO - NAT COMMUN
JF - NAT COMMUN
SN - 2041-1723
IS - 7933
ER -