In situ observation of cell-detachment process initiated by femtosecond laser-induced stress wave

Maezawa, Yasuyo; Hosokawa, Yoichiroh; Okano, Kazunori; Matsubara, Mie; Masuhara, Hiroshi

doi:10.1007/s00339-010-5771-4

Cited by 9 publications

(8 citation statements)

References 13 publications

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“…Previously, the cell detachment process from the source substrate due to trypsion treatment has been compared with that due to the impulsive force. This comparison was performed by visualizing a collagen scaffold on the source substrate with a fluorescence probe of quantum dots (Maezawa et al 2010). When cell was detached from the substrate by the impulsive force loading, fluorescence from the scaffold simultaneously disappeared from the substrate, although the scaffold was not irradiated by the laser.…”

Section: Resultsmentioning

confidence: 99%

Morphological evaluation of cell differentiation after the isolation of single cells by a femtosecond laser-induced impulsive force

Maezawa

Okano

Matsubara

et al. 2010

Biomed Microdevices

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When nerve growth factor (NGF) is interacted with PC12 cells derived from rat pheochromocytoma, they are partially differentiated into neuron-like cells with neurites. In this work, PC12 cells differentiated by NGF were selectively isolated using a localized impulsive force in a μm-scale area, which was generated by focusing an infrared femtosecond laser into a cell culture medium. In order to evaluate the ability of the isolation method, differentiated and undifferentiated cells were isolated and their morphological changes after the isolation were compared. In both cases, their neurites were once contracted and some of them gradually regenerated day by day. When differentiated cells were isolated, the percentage of differentiated cells with regenerated neurites, 6 h after the isolation, was about 3.3 times higher than that when undifferentiated ones were isolated. This result was compared with a control trypsin experiment. In the comparison, it was indicated that the same degree of cell function was maintained when the present isolation method was used.

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Section: Resultsmentioning

confidence: 99%

Morphological evaluation of cell differentiation after the isolation of single cells by a femtosecond laser-induced impulsive force

Maezawa

Okano

Matsubara

et al. 2010

Biomed Microdevices

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“…[20] This value is comparable to, or higher than, that for cells released by trypsin. [21] In this study, we showed that detached cells are sequentially translocated to adjacent cell-adhesive domains on the R f platform in a few minutes by applying the stress wave as illustrated in Figure 1 A. When the fs laser is focused on the platform, the glass absorbs the photons, and this is followed within a short time by an ablation sequence.…”

Section: Application Of Fs Laser In Cell Arraying and In Situ Lithogrmentioning

confidence: 97%

“…Our group has demonstrated that the stress wave could detach cells individually from a culture substrate. [19][20][21] Since the force affecting the cells was estimated to be of the order of just nano-Newtons, [19] cells (e.g., mouse embryonic NIH3T3) would readhere to the substrate in high yield (80 %). [20] This value is comparable to, or higher than, that for cells released by trypsin.…”

Section: Application Of Fs Laser In Cell Arraying and In Situ Lithogrmentioning

confidence: 99%

Induction of Cell–Cell Connections by Using in situ Laser Lithography on a Perfluoroalkyl‐Coated Cultivation Platform

Okano

Matsui

et al. 2011

ChemBioChem

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This article describes a novel laser-directed microfabrication method carried out in aqueous solution for the organization of cell networks on a platform. A femtosecond (fs) laser was applied to a platform culturing PC12, HeLa, or normal human astrocyte (NHA) cells to manipulate them and to facilitate mutual connections. By applying an fs-laser-induced impulsive force, cells were detached from their original location on the plate, and translocated onto microfabricated cell-adhesive domains that were surrounded with a cell-repellent perfluoroalkyl (R(f)) polymer. Then the fs-laser pulse-train was applied to the R(f) polymer surface to modify the cell-repellent surface, and to make cell-adhesive channels of several μm in width between each cell-adhesive domain. PC12 cells elongated along the channels and made contact with others cells. HeLa and NHA cells also migrated along the channels and connected to the other cells. Surface analysis by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) confirmed that the R(f) polymer was partially decomposed. The method presented here could contribute not only to the study of developing networks of neuronal, glial, and capillary cells, but also to the quantitative analysis of nerve function.

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“…When a near-infrared femtosecond laser is tightly focused in an aqueous solution under a microscope, shock and stress waves are generated at the laser focal point (4)(5)(6). These waves localize in a micrometer-sized space around the laser focal point (4) and they act as an impulsive force on a micrometer-sized object, such as a single cell, which is located near the laser focal point (7)(8)(9)(10). We previously succeeded in quantifying the magnitude of the micrometer-sized impulsive force using atomic force microscopy (AFM) (11,12).…”

Section: Introductionmentioning

confidence: 99%

Time-Course Statistical Evaluation of Intercellular Adhesion Maturation by Femtosecond Laser Impulse

Iino

Hagiyama

Furuno

et al. 2016

Biophysical Journal

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The maturation of intercellular adhesion is an essential process for establishing the signal transduction network in living cells. Although the maturation is naturally considered to enhance the signal transduction, the relationship between the signal transduction and the maturation process has not been revealed in detail using time-course data. Here, using a coculture of mast cells and neurites, differences in maturation between individual cells were estimated as a function of the adhesion strength by our original single-cell measurement method utilizing a laser-induced impulsive force. When an intense femtosecond laser is focused into a culture medium under a microscope, shock and stress waves are generated at the laser focal point that exert an impulsive force on individual cells. In our method, this impulse is used to break the adhesion between a mast cell and a neurite. The magnitude of the impulse is then quantified by a local force-measurement system utilizing an atomic force microscope, and the adhesion strength is estimated from the threshold of the impulse required to break the adhesion. The measurement is conducted within 1 min/cell, and thus, data on the individual differences of the adhesion strength can be obtained within only a few hours. Coculturing of neurites and mast cells for 4 h resulted in a specific adhesion that was stronger than the nonspecific adhesions between the substrate and mast cells. In the time-course investigation, we identified two distinct temporal patterns of adhesion: 1) the strength at 24 h was the same as the initial strength; and 2) the strength increased threefold from baseline and became saturated within 24 h. Based on these results, the distribution of CADM1 adhesion molecules in the neurites was suggested to be inhomogeneous, and the relationship between adhesion maturation and the signal-transduction process was considered.

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In situ observation of cell-detachment process initiated by femtosecond laser-induced stress wave

Cited by 9 publications

References 13 publications

Morphological evaluation of cell differentiation after the isolation of single cells by a femtosecond laser-induced impulsive force

Morphological evaluation of cell differentiation after the isolation of single cells by a femtosecond laser-induced impulsive force

Induction of Cell–Cell Connections by Using in situ Laser Lithography on a Perfluoroalkyl‐Coated Cultivation Platform

Time-Course Statistical Evaluation of Intercellular Adhesion Maturation by Femtosecond Laser Impulse

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