Chromosome Lesions Produced with an Argon Laser Microbeam without Dye Sensitization

Berns, Michael W.; Cheng, Wanny K.; Floyd, Alton D.; Ohnuki, Yasushi

doi:10.1126/science.171.3974.903

Cited by 42 publications

(14 citation statements)

References 4 publications

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“…Previous phase-contrast microscope studies involving microirradiation with the 225–300 nm UV laser (39,40), 488–514 nm argon ion laser (21,23) and the 483–519 nm dye laser (21,36) have described chromosomal alterations as phase-paled regions when compared with the surrounding chromatin. These early studies were performed with laser exposures in the millisecond (ms) and microsecond (µs) domains.…”

Section: Discussionmentioning

confidence: 99%

Analysis of DNA double-strand break response and chromatin structure in mitosis using laser microirradiation

Gomez-Godinez¹,

Wu²,

Sherman³

et al. 2010

Nucleic Acids Research

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In this study the femtosecond near-IR and nanosecond green lasers are used to induce alterations in mitotic chromosomes. The subsequent double-strand break responses are studied. We show that both lasers are capable of creating comparable chromosomal alterations and that a phase paling observed within 1–2 s of laser exposure is associated with an alteration of chromatin as confirmed by serial section electron microscopy, DAPI, γH2AX and phospho-H3 staining. Additionally, the accumulation of dark material observed using phase contrast light microscopy (indicative of a change in refractive index of the chromatin) ∼34 s post-laser exposure corresponds spatially to the accumulation of Nbs1, Ku and ubiquitin. This study demonstrates that chromosomes selectively altered in mitosis initiate the DNA damage response within 30 s and that the accumulation of proteins are visually represented by phase-dark material at the irradiation site, allowing us to determine the fate of the damage as cells enter G1. These results occur with two widely different laser systems, making this approach to study DNA damage responses in the mitotic phase generally available to many different labs. Additionally, we present a summary of most of the published laser studies on chromosomes in order to provide a general guide of the lasers and operating parameters used by other laboratories.

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Section: Discussionmentioning

confidence: 99%

Analysis of DNA double-strand break response and chromatin structure in mitosis using laser microirradiation

Gomez-Godinez¹,

Wu²,

Sherman³

et al. 2010

Nucleic Acids Research

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“…Time-resolved monitoring of laserinduced cell damage clearly confirmed that for both pigmented and non-pigmented cells the damage processes can be detected and evaluated in similar way. Comparisons of damage thresholds for strongly absorbent cells, such as RBCs (0.5-1 J/cm 2 at a 420-nm wavelength with a hemoglobin absorption coefficient of around 1,100-1,300 cm À1 [23]), with data obtained with pigmented retinal cells (0.1-0.8 J/cm 2 at 694 nm and 532 nm wavelengths, with a melanin absorption coefficient of around 1,000-1,900 cm À1 [2,5,7]) show similar threshold parameters. There are no available data for comparisons with other non-pigmented cells.…”

Section: Discussionmentioning

confidence: 99%

Spectral evaluation of laser-induced cell damage with photothermal microscopy

Lapotko

Zharov

2005

Lasers Surg. Med.

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“…1 Through their continual refinement, a high level of precision in single-cell manipulation has been achieved. Noteworthy cellular applications of optical scissors include the ablation of axons, 2 microtubules, 3,4 and chromosomes, [5][6][7] as well as plasma membrane poration. 8 Recently, optical scissors have seen extensive use in DNA damage repair research by exposing a linear path across the nucleus of interphase cells followed by the monitoring of DNA damage recognition and repair in the altered region.…”

Section: Introductionmentioning

confidence: 99%

Spatially sculpted laser scissors for study of DNA damage and repair

et al. 2009

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Abstract. We present a simple and efficient method for controlled linear induction of DNA damage in live cells. By passing a pulsed laser beam through a cylindrical lens prior to expansion, an elongated elliptical beam profile is created with the ability to expose controlled linear patterns while keeping the beam and the sample stationary. The length and orientation of the beam at the sample plane were reliably controlled by an adjustable aperture and rotation of the cylindrical lens, respectively. Localized immunostaining by the DNA double strand break ͑DSB͒ markers phosphorylated H2AX ͑␥H2AX͒ and Nbs1 in the nuclei of HeLa cells exposed to the "line scissors" was shown via confocal imaging. The line scissors method proved more efficient than the scanning mirror and scanning stage methods at induction of DNA DSB damage with the added benefit of having a greater potential for high throughput applications.

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Chromosome Lesions Produced with an Argon Laser Microbeam without Dye Sensitization

Abstract: Improvements in the argon laser microbeam have made it possible to cause damage to chromosomes of tissue culture cells without prior treatment of the cells with a photosensitizing agent. These results have been confirmed independently in two laboratories.

Cited by 42 publications

References 4 publications

Analysis of DNA double-strand break response and chromatin structure in mitosis using laser microirradiation

Analysis of DNA double-strand break response and chromatin structure in mitosis using laser microirradiation

Spectral evaluation of laser-induced cell damage with photothermal microscopy

Spatially sculpted laser scissors for study of DNA damage and repair

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