Intracellular disruption of mitochondria in a living HeLa cell with a 76-MHz femtosecond laser oscillator

Shimada, Toshio; Watanabe, Wataru; Matsunaga, Sachihiro; Higashi, Taishi; Ishii, Hiroshi; Fukui, Kiichi; Isobe, Keisuke; Itoh, Kazuyoshi

doi:10.1364/opex.13.009869

Cited by 66 publications

(37 citation statements)

References 32 publications

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“…Furthermore, when applied to plant cells, laser nanosurgery gives the possibility to dissect the cell wall in living tissues (Tirlapur and Konig , 2002a ) individual chloroplasts without compromising the cell viability. This fi nding paved the way for the ablation of other membrane bound organelles such as mitochondria in mammalian cultured cells (Figure 2 B; Khodjakov et al , 2004b ;Watanabe et al , 2004 ;Shen et al , 2005 ;Shimada et al , 2005 ).…”

Section: Membranes and Membrane-bound Organellesmentioning

confidence: 94%

“…Furthermore, the size and positioning of membrane bound organelles could be studied after partial laser-mediated removal. As discussed before, different studies showed the possibility to ablate chloroplasts and mitochondria (Tirlapur and Konig , 2002a ;Watanabe et al , 2004 ;Shen et al , 2005 ;Shimada et al , 2005 ) but their subsequent biogenesis and spatial re-organization has not yet been addressed. Given the fact that organelle localization is emerging as a crucial determinant for signaling events and activation of different pathways (Korolchuk et al , 2011 ), research on organelle shape and spatial organization as well as their biogenesis could take advantage of laser nanosurgery and micropatterning to gain insights into topics that are otherwise diffi cult to address.…”

Section: Laser Nanosurgery On Patterned Cells: a Promising Applicationmentioning

confidence: 99%

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At the cutting edge: applications and perspectives of laser nanosurgery in cell biology

Ronchi

Terjung

Pepperkok³

2012

BCHM

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Laser-mediated nanosurgery has become popular in the last decade because of the previously unexplored possibility of ablating biological material inside living cells with sub-micrometer precision. A number of publications have shown the potential applications of this technique, ranging from the dissection of sub-cellular structures to surgical ablations of whole cells or tissues in model systems such as Drosophila melanogaster or Danio rerio . In parallel, the recent development of micropatterning techniques has given cell biologists the possibility to shape cells and reproducibly organize the intracellular space. The integration of these two techniques has only recently started yet their combination has proven to be very interesting. The aim of this review is to present recent applications of laser nanosurgery in cell biology and to discuss the possible developments of this approach, particularly in combination with micropattern-mediated endomembrane organization.

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Section: Membranes and Membrane-bound Organellesmentioning

confidence: 94%

Section: Laser Nanosurgery On Patterned Cells: a Promising Applicationmentioning

confidence: 99%

At the cutting edge: applications and perspectives of laser nanosurgery in cell biology

Ronchi

Terjung

Pepperkok³

2012

BCHM

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“…Focused femtosecond laser has been successfully used to dissect single dendrites and cytoskeletons, to optically knock out intracellular mitochondria [2][3][4], for membrane and microtubules surgery [5], for chromosome dissection [6] as well as for ocular refractive surgery [7]. Femtosecond laser pulses have been also used for efficient targeted transfection of Chinese hamster ovary cells [8] and canina mammary cells MTH53a [9] by transient opening of the cellular membrane and the subsequent diffusion of the foreign DNA into the cytoplasm.…”

Section: Biophotonicsmentioning

confidence: 99%

Optical knock out of stem cells with extremely ultrashort femtosecond laser pulses

Uchugonova

Isemann²,

Gorjup

et al. 2008

Journal of Biophotonics

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Novel ultracompact multiphoton. sub-20 femtosecond near infrared 85MHz laser scanning microscopes and conventional 250 fs laser microscopes have been used to perform, high spatial resolution two-photon imaging of stem cell clusters as well, as selective-,int.l.lacellula,r nanoprocessing and knock out of living single stem cells within an 3D microenvironment without any collateral damage. Also lethal, cell exposure of large, parts of cell clusters was successfully probed while maintaining single cells of interest alive. The mew power could be kept in the milliwatt range for 3D nanoprocessing and even in the microwatt range for two-photon imabing. Ultracompact low power sub-20 fs laser systems may becone interesting tools for optical nanobiotechnology such as optical cleaning of stem clusters as well as optical transfection. [GRAPHICS] Optical knock out of a single human pancreatic stem cell without collateral damage

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Section: B the Middle Years: Laser-based Microirradiationmentioning

confidence: 99%

“…In fact, it appears that punching holes in mitochondria with new and improved lasers, without providing any additional insight into how these organelles work, has become a litmus test for proving the utility of a new system. Just last year there were at least three independent reports of this exact experiment (Colombelli et al, 2005;Shen et al, 2005;Shimada et al, 2005).Initially, the complexity of laser microirradiation systems restricted their distribution to just a few institutions specializing primarily in laser physics. However, in early 1980s Michael Berns started the laser microbeam program (LAMP) at the University of California (Irvine) which was (and remains) sponsored by the NIH Center for Research Resources as a National Biotechnology Resource.…”

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confidence: 99%

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

Magidson

Lončarek

Hergert

et al. 2007

Methods in Cell Biology

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Modern biology is based largely on a reductionistic "dissection" approach-most cell biologists try to determine how complex biological systems work by removing their individual parts and studying the effects of this removal on the system. A variety of enzymatic and mechanical methods have been developed to dissect large cell assemblies like tissues and organs. Further, individual proteins can be inactivated or removed within a cell by genetic manipulations (e.g., RNAi or gene knockouts). However, there is a growing demand for tools that allow intracellular manipulations at the level of individual organelles. Laser microsurgery is ideally suited for this purpose and the popularity of this approach is on the rise among cell biologists. In this chapter, we review some of the applications for laser microsurgery at the subcellular level and describe practical requirements for laser microsurgery instrumentation demanded in the field. We also outline a relatively inexpensive but versatile laser microsurgery workstation that is being used in our laboratory. Our major thesis is that the limitations of the technology are no longer at the level of the laser, microscope, or software, but instead only in defining creative questions and in visualizing the target to be destroyed.At last in an incredible manner he [Archimedes] burned up the whole Roman fleet. For by tilting a kind of mirror toward the sun he concentrated the sun's beam upon it; and owing to the thickness and smoothness of the mirror he ignited the air from this beam and kindled a great flame, the whole of which he directed upon the ships that lay at anchor in the path of the fire, until he consumed them all.1 I. History of the Field A. The Genesis of "Micro-Photo-Surgery"The origins of Cell Biology as a branch of "natural philosophy" can be traced to the English polymath Robert Hooke who, using a hand-crafted, leather and goldtooled compound light microscope (LM), published a book containing elaborate drawings of magnified objects which he called Micrographia. In this book, which became an immediate best-seller (and has been reprinted countless times), Hooke used the term "cell" to describe the repeating units seen in magnified slices of cork that resembled the monk cells of a monastery. Ironically, these repeating units were not actual cells but rather just the cellulose walls that surround cells in plant tissues. It took another 175 years of optical development and exploration, before Schleiden and Schwann (1839) convincingly asserted that cells are the fundamental building blocks of all life. In 1855, the Prussian physician and politician Virchow postulated that cells arise only from preexisting cells by reproduction and cannot be formed de novo from amorphous "living matter" or "protoplasma." This principle, which Virchow eloquently 1 Dio's Roman History, translated by Earnest Cary Loeb Classical Library, Harvard University Press, Cambridge, 1914, Vol. II, p. 171. NIH Public Access Author ManuscriptMethods Cell Biol. Author manuscript; availabl...

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Intracellular disruption of mitochondria in a living HeLa cell with a 76-MHz femtosecond laser oscillator

Cited by 66 publications

References 32 publications

At the cutting edge: applications and perspectives of laser nanosurgery in cell biology

At the cutting edge: applications and perspectives of laser nanosurgery in cell biology

Optical knock out of stem cells with extremely ultrashort femtosecond laser pulses

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

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