Mechanisms of femtosecond laser nanosurgery of cells and tissues

Abstract

We report on femtosecond nanosurgery of fluorescently labeled structures in cells with a spatially superresolved laser beam. The focal spot width is reduced using phase filtering applied with a programmable phase modulator. A comprehensive statistical analysis of the resulting cuts demonstrates an achievable average resolution enhancement of 30 %.

Spatially, Temporally, and Quantitatively Controlled Delivery of Broad Range of Molecules into Selected Cells through Plasmonic Nanotubes

“…[ 21 ] Among them, it is well known that they are very effective in being injected into a semiconductor material. [22][23][24] In laser nanosurgery, water is usually treated as an amorphous semiconductor [ 25,26 ] in which free-electron density can be generated by exciting electrons from the valence to the conduction band. This process requires activation energy equal to 6.5 eV, whereas the energy necessary to remove an electron from gold in the presence of water is 3.7-2.2 eV, [27][28][29] thus making the plasmonic process strongly favored and effi cient.…”

Application of dynamic diffractive optics for enhanced femtosecond laser based cell transfection

“…The cavitation bubbles caused by the low plasma formation are typically short lived and smaller than the resolving power of the objectives [33] However, if the dose is sufficiently high, a visible bubble in brightfield imaging can be observed. We did not see any optoinjection in the absence of visible bubbles, which confirms a similar observation reported by Baumgart et al [26].…”

Biophysical Response to Pulsed Laser Microbeam‐Induced Cell Lysis and Molecular Delivery

“…This enables the generation of controlled and reproducible regions of cellular damage and/or molecular delivery. Moreover, with the potential development of laser microbeam platforms that offer multiple wavelengths (e.g., λ = 355,532,1064nm) and a range of pulse durations (ns-fs) that can provide for optical breakdown over a range of pulse energies [41,22], one can easily conceive of a single platform that can provide for precise zones of cellular damage and molecular delivery over a broad range.…”

“…As a result, there is an increasing interest to use pulsed laser microbeams for precise cellular manipulation, including laserinduced cell lysis [1], cell microdissection and catapulting [2][3][4][5], cell collection, expansion, and purification [6][7][8], cellular microsurgery [9][10][11], and cell membrane permeabilization for the delivery of membrane-impermeant molecules into cells [12 15], The processes of laser-induced optoinjection and optoporation offer the ability to load cells with a variety of biomolecules on short time scales (milliseconds to seconds) through optically produced cell membrane permeabilization [12,14,15], Despite the innovative utilization of laser microbeams in cell biology and biotechnology, only recently have studies provided insight regarding the mechanisms that mediate the interactions of highly focused pulsed laser beams with cells [16][17][18][19][20][21][22], A better understanding of these processes will prove critical to the continued development of laser microbeams for both research and practical applications. In previous studies, we provided a detailed characterization of the physics involved in the interaction of highly-focused nanosecond laser microbeams with cells [19,20], However, it is important to relate these physical effects to the biological response of the cells.…”