Research Papers

Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope

[+] Author Affiliations
Leonardo Sacconi

University of Trento, via Sommarive 14, 38050 Povo (TN), Italy

Iva M. Tolic´-Nørrelykke

European Laboratory for Non-linear Spectroscopy, via Nello Carrara 1, 50019 Sesto Fiorentino (Fl), Italy

Rugjer Bos?kovic´ Institute, Bijenic?ka 54, 10000 Zagreb, Croatia

Renzo Antolini

University of Trento, via Sommarive 14, 38050 Trento, Italy

Francesco S. Pavone

European Laboratory for Non-linear Spectroscopy, via Nello Carrara 1, 50019 Florence, Italy

University of Florence and INFM Sezione di Firenze, Department of Physics, via Sansone 1, 50019 Florence, Italy E-mail: pavone@lens.unifi.it

J. Biomed. Opt. 10(1), 014002 (Feb. 3, 2005). doi:10.1117/1.1854675
History: Received Mar. 1, 2004; Revised May 27, 2004; Accepted May 28, 2004; Feb. 3, 2005; Online February 03, 2005
Text Size: A A A

We use near-IR femtosecond laser pulses for a combination of microscopy and nanosurgery on fluorescently labeled structures within living cells. Three-dimensional reconstructions of microtubule structures tagged with green fluorescent protein (GFP) are made during different phases of the cell cycle. Further, the microtubules are dissected using the same laser beam but with a higher laser power than for microscopy. We establish the viability of this technique for the cells of a fission yeast, which is a common model to study the mechanics of cell division. We show that nanosurgery can be performed with submicrometer precision and without visible collateral damage to the cell. The energy is primarily absorbed by the GFP molecules, and not by other native structures in the cell. GFP is particularly suitable for multiphoton excitation, as its excitation wavelength near 900 nm is benign for most cellular structures. The ability to use GFP to label structures for destruction by multiphoton excitation may be a valuable tool in cell biology. © 2005 Society of Photo-Optical Instrumentation Engineers.

© 2005 Society of Photo-Optical Instrumentation Engineers

Citation

Leonardo Sacconi ; Iva M. Tolic´-Nørrelykke ; Renzo Antolini and Francesco S. Pavone
"Combined intracellular three-dimensional imaging and selective nanosurgery by a nonlinear microscope", J. Biomed. Opt. 10(1), 014002 (Feb. 3, 2005). ; http://dx.doi.org/10.1117/1.1854675


Tables

Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

Advertisement


  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.