Laser Microsurgery in Cell and Developmental Biology

Abstract

New applications of laser microbeam irradiation to cell and developmental biology include a new instrument with a tunable wavelength (217- to 800-nanometer) laser microbeam and a wide range of energies and exposure durations (down to 25 × 10 -12 second). Laser microbeams can be used for microirradiation of selected nucleolar genetic regions and for laser microdissection of mitotic and cytoplasmic organelles. They are also used to disrupt the developing neurosensory appendages of the cricket and the imaginal discs of Drosophila .

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

“…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.…”

Laser Microsurgery in the GFP Era: A Cell Biologist's Perspective

“…As emphasized early on by Berns et al (1981), Gaussian laser beams can actually generate a central "hot spot" inside the Airy disk. Thus, it is possible to select a beam energy at which damage to the specimen is restricted only to the "hot spot" at the peak intensity in the center of the Airy disk.…”

“…During the next several years this workstation was used to demonstrate, for example, that the spindle assembly checkpoint monitors kinetochore attachment , that chromosomes containing a single kinetochore can congress to the equator of the forming spindle (Khodjakov et al, 1997a), and that entry into mitosis in vertebrate cells is guarded by a DNA damage checkpoint that reverses the cell cycle when triggered during early prophase (Rieder and Cole, 1998). Further, this same instrument was also used by other biologists to address a number of questions in systems as diverse as fungi to cranefly (Inoue et al, 1998;LaFountain, Jr., et al, 2001, 2002Orokos et al, 2000).As emphasized early on by Berns et al (1981), Gaussian laser beams can actually generate a central "hot spot" inside the Airy disk. Thus, it is possible to select a beam energy at which damage to the specimen is restricted only to the "hot spot" at the peak intensity in the center of the Airy disk.…”

Measurement of Tension Release During Laser Induced Axon Lesion to Evaluate Axonal Adhesion to the Substrate at Piconewton and Millisecond Resolution

“…With the exploitation of effects such as radiation pressure (as in optical tweezers 1 ), or high-energy photon flux (as in laser dissector 2 ), this technology was extended to nano-manipulation. The optical imaging system furnishes a precise control to visualize and manipulate sub cellular targets 3 .…”