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Pregled bibliografske jedinice broj: 898299

Dissection and characterization of microtubule bundles in the mitotic spindle using femtosecond laser ablation


Buđa, Renata; Vukušić, Kruno; Tolić, Iva Marija
Dissection and characterization of microtubule bundles in the mitotic spindle using femtosecond laser ablation // Methods in cell biology, 139 (2017), 81-101 doi:10.1016/bs.mcb.2016.11.007 (međunarodna recenzija, članak, znanstveni)


Naslov
Dissection and characterization of microtubule bundles in the mitotic spindle using femtosecond laser ablation

Autori
Buđa, Renata ; Vukušić, Kruno ; Tolić, Iva Marija

Izvornik
Methods in cell biology (0091-679X) 139 (2017); 81-101

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
Bridging fiber, Confocal laser scanning microscopy, Forces, k-fiber, Laser cutting, Metaphase, Microtubules, Mitosis, Swept field confocal microscopy

Sažetak
The mitotic spindle is a highly organized and dynamic structure required for segregation of the genetic material into two daughter cells. Although most of the individual players involved in building the spindle have been characterized in vitro, a general understanding of how all of the spindle players act together in vivo is still missing. Hence, in recent years, experiments have focused on introducing mechanical perturbations of the spindle on a micron scale, thereby providing insight into its function and organization, as well as into forces acting in the spindle. Among different types of mechanical perturbations, optical ones are more flexible, less invasive, and more precise than other approaches. In this chapter, we describe a detailed protocol for cutting the microtubule bundles in human cells using a near-infrared femtosecond laser. This type of laser microsurgery provides the ability to precisely sever a single microtubule bundle while preserving spindle integrity and dynamics. Furthermore, we describe quantitative measurements obtained from the response of a severed microtubule bundle to laser ablation, which reveal the structure and function of individual parts of the spindle, such as the bridging fiber connecting sister k-fibers. Finally, the method described here can be easily combined with other quantitative techniques to address the complexity of the spindle.

Izvorni jezik
Engleski

Znanstvena područja
Biologija



POVEZANOST RADA


Projekt / tema
EK-H2020-647077 (Iva TOLIC, EK - ERC-2014-CoG)

Ustanove
Institut "Ruđer Bošković", Zagreb

Autor s matičnim brojem:
Iva Marija Tolić, (260543)

Časopis indeksira:


  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus
  • MEDLINE


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