Formation of light-absorbing centers induced in cytoplasm of mouse embryos by femtosecond pulsed near-infrared radiation

Астафьев, А. А.; Zalesskii, A. D.; Shakhov, Alexander N.; Osychenko, A. A.; Надточенко, В. А.

doi:10.1134/s0018143916050039

Cited by 6 publications

(2 citation statements)

References 8 publications

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“…Similar effects of accumulation of carbonized products in biological species upon ultrashort laser irradiation have been observed previously . They also explain emergence of near-infrared (NIR) absorption in the laser-irradiated cytoplasm which we previously found in the mouse embryo and also observed in the present experiments, because unlike biological materials, carbonaceous species produced by laser irradiation can exhibit an appreciable absorption in the NIR region. Analysis of the Raman spectra reveals additional characteristics of the carbonized species.…”

Section: Results and Discussionmentioning

confidence: 99%

Probing Intracellular Dynamics Using Fluorescent Carbon Dots Produced by Femtosecond Laser In Situ

et al. 2020

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Fluorescent particle tracking is a powerful technique for studying intracellular transport and microrheological properties within living cells, which in most cases employs exogenous fluorescent tracer particles delivered into cells or fluorescent staining of cell organelles. Herein, we propose an alternative strategy, which is based on the generation of fluorescent species in situ with ultrashort laser pulses. Using mouse germinal vesicle oocytes as a model object, we demonstrate that femtosecond laser irradiation produces compact dense areas in the intracellular material containing fluorescent carbon dots synthesized from biological molecules. These dots have tunable persistent and excitation-dependent emission, which is highly advantageous for fluorescent imaging. We further show that tight focusing and tuning of irradiation parameters allow precise control of the location and size of fluorescently labeled areas and minimization of damage inflicted to cells. Pieces of the intracellular material down to the submicrometer size can be labeled with laser-produced fluorescent dots in real time and then employed as probes for detecting intracellular motion activity via fluorescent tracking. Analyzing their diffusion in the oocyte cytoplasm, we arrive to realistic characteristics of active forces generated within the cell and frequency-dependent shear modulus of the cytoplasm. We also quantitatively characterize the level of metabolic activity and density of the cytoskeleton meshwork. Our findings establish a new technique for probing intracellular mechanical properties and also promise applications in tracking individual cells in population or studies of spatiotemporal cell organization.

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Section: Results and Discussionmentioning

confidence: 99%

Probing Intracellular Dynamics Using Fluorescent Carbon Dots Produced by Femtosecond Laser In Situ

et al. 2020

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“…It was reported that irradiation with pulsed laser light can induce synthesis of CDs from an aromatic precursor possibly through a plasma-mediated mechanism [31]. While there were evidences of femtosecond laser-induced carbonization in biological tissues [32][33][34] and formation of light absorbing centers [35] a laser synthesis of carbon dots from bio-molecules has not been positively demonstrated.…”

Section: Introductionmentioning

confidence: 99%

Live cell bioimaging with carbon dots produced in situ by femtosecond laser from intracellular material

Shakhov

Astafiev

Osychenko

et al. 2018

Preprint

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Owning to excellent optical properties and high bio-compatibility carbon dots (CDs) have drawn increasing attention and have been widely applied as imaging agents for various bio-applications. Here we report a strategy for live-cell fluorescent bioimaging based on in situ synthesis of CDs within cells by tightly focused femtosecond laser pulses. Laser-produced carbon dots exhibit bright excitationdependent fluorescence and are highly two-photon active under near infrared femtosecond excitation, thus demonstrating a potential for two-photon fluorescence imaging. The Raman spectra of fluorescent centers show strong D (1350 cm -1 ) and G (1590 cm -1 ) bands, thus suggesting that they are composed of carbon dots with sp 2 -hybridyzed core. Using Mouse GV oocytes as a model system we examine cytotoxicity and demonstrate the possibility of long-term fluorescent intracellular tracking of the laser-produced CDs. Created virtually in any point of the live cell, CD-based fluorescent µm-sized markers demonstrate high structural stability and retain bright fluorescence many hours after formation. Our results point to laser-produced fluorescent CDs as a highly-potent tool for cell cycle tracking, culture cell marking and probing intracellular movements.

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The use of optical chopper increases the efficiency of femtosecond laser-induced cell fusion

Osychenko

Zalessky

Tochilo

et al. 2020

J. Phys.: Conf. Ser.

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Artificial cell fusion is a widely used approach in cell biology and biomedicine. Femtosecond laser-induced cell fusion is considered to be a precise and low-invasive tool for the cell fusion. However, the percentage of somatic cell fusion remains not very high, and the use of polyethylene glycol is often required. In our research we propose the use of rotating optical chopper to produce ultra-short trains of femtosecond pulses (up to 1 ms) for decreasing the laser impact. It helps to control the appearance and size of gas-vapor bubbles, avoiding cell destruction. We achieved the cell fusion efficiency of 45% without the use of polyethylene glycol, but only in freshly thawed cells. Continuously cultured cells completely failed to fuse.

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Formation of light-absorbing centers induced in cytoplasm of mouse embryos by femtosecond pulsed near-infrared radiation

Cited by 6 publications

References 8 publications

Probing Intracellular Dynamics Using Fluorescent Carbon Dots Produced by Femtosecond Laser In Situ

Probing Intracellular Dynamics Using Fluorescent Carbon Dots Produced by Femtosecond Laser In Situ

Live cell bioimaging with carbon dots produced in situ by femtosecond laser from intracellular material

The use of optical chopper increases the efficiency of femtosecond laser-induced cell fusion

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