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Then the cells were washed with normal media and incubated in the humidified 5% CO 2 incubator at 37 ◦ C.

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After attenuation through prisms and filters, the power of the laser exposed on the samples was only about 2 m W, which made it almost impossible for the laser to degrade the biological samples.

Spectra were collected from cells with 50 × long-working-distance objective (NA 0.5, Olympus, Japan), and the signal was integrated for 2 × 60 s and measured over a spectral range of 700–1800 cm − 1 with respect to the excitation wavenumber.

5 × 10 8 cells m L − 1 , as used for Raman measurements.

Working solutions of SNP were prepared by dissolving the yellow powder with PBS.

Confocal laser scanning microscopy (CLSM) is the most popular, nondestructive technique used to monitor the time-resolved biochemical changes in a single cell.

In this technique, the different targeting biomacromolecules can be identified and quantitatively visualized by the addition of fluorescent probes.

In addition, Raman spectroscopy was also demonstrated to be more sensitive for in vivo and in situ monitoring of the drug stimulus response of single cells than conventional flow cytometric assay, especially at low drug concentration.

Human oral squamous cell carcinoma cells (TCA8113) were purchased from the China Center for Typical Culture Collection (CCTCC).

Sodium nitroprusside (SNP), a drug used for angiectasis, acted as a NO source by the spontaneous release of NO in solution.

The amount of NO could be controlled by adjusting the SNP concentration in culture solution.

Cell suspensions were then centrifuged into a pellet (1000 rpm for 4 min) and the medium was removed.

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