We have developed a versatile technique for ultra-high-precision laser machining that uses tightly focused femtosecond laser pulses to ablate sharply defined nanometer-scale regions in materials. With this technology it is possible to selectively ablate regions even smaller than 20 nm across. This milestone achievement holds great promise for a wide range of applications, perhaps none more exciting than as a tool to highly selectively destroy intracellular structures. The ability to create structural """"""""knockouts"""""""" in which intracellular components are selectively destroyed holds enormous promise for elucidating structure-function relationships, just as molecular-genetic knockouts have been crucial to understanding the function of genes and the proteins they encode. Toward fulfilling this potential, this proposal has dual goals of demonstrating the utility of this approach to the broader biological community, and addressing fundamental questions concerning cell division. To these ends we propose to apply structural knockout technology to study the biomechanics of the cytoskeleton and mitosis. These experiments will characterize: 1) the mechanical and force generating properties that allow chromosomes to bind and move along microtubules, 2) the antimitotic activity of the chemotherapy drug taxol, and 3) the role of centrioles in establishing mitotic architecture. The ultimate limits of structural knockout technology will also be investigated to explore the potential for intriguing future applications such as targeted disruption of single molecules or genes.
| Cheng, Jun; Tiyaboonchai, Amita; Yamashita, Yukiko M et al. (2011) Asymmetric division of cyst stem cells in Drosophila testis is ensured by anaphase spindle repositioning. Development 138:831-7 |
| Lee, Sanghyun; An, Ran; Hunt, Alan J (2010) Liquid glass electrodes for nanofluidics. Nat Nanotechnol 5:412-6 |
| Herbstman, Jeffrey F; Hunt, Alan J (2010) High-aspect ratio nanochannel formation by single femtosecond laser pulses. Opt Express 18:16840-8 |
| Cheng, Jun; Waite, Andrea L; Tkaczyk, Eric R et al. (2010) Kinetic properties of ASC protein aggregation in epithelial cells. J Cell Physiol 222:738-47 |
| Yamashita, Yukiko M; Yuan, Hebao; Cheng, Jun et al. (2010) Polarity in stem cell division: asymmetric stem cell division in tissue homeostasis. Cold Spring Harb Perspect Biol 2:a001313 |
| Cheng, Jun; Hunt, Alan J (2009) Time-lapse live imaging of stem cells in Drosophila testis. Curr Protoc Stem Cell Biol Chapter 2:Unit 2E.2 |
| Ke, Kevin; Cheng, Jun; Hunt, Alan J (2009) The distribution of polar ejection forces determines the amplitude of chromosome directional instability. Curr Biol 19:807-15 |
| An, Ran; Uram, Jeffrey D; Yusko, Erik C et al. (2008) Ultrafast laser fabrication of submicrometer pores in borosilicate glass. Opt Lett 33:1153-5 |
| Herbstman, Jeffrey F; Hunt, Alan J; Yalisove, Steven M (2008) Novel morphologies and non-linear scaling of laser damage in glass by tightly-focused femtosecond pulses. Appl Phys Lett 93:11112 |
| Cheng, Jun; Turkel, Nezaket; Hemati, Nahid et al. (2008) Centrosome misorientation reduces stem cell division during ageing. Nature 456:599-604 |
