Nanoscale scanning electrochemical microscopy is emerging as a powerful electrochemical technique due to recent pioneering advances in tip fabrication and handling, instrument and cell design, and awareness of airborne and solution organic contamination which impact tip and substrate handling.
After two washes in PBSA, the cells were resuspended in 0.5% paraformaldehyde and counted using a FACScan instrument and Cell Quest software (Becton Dickinson).
Then, the absorbance was measured at a wavelength of 682 nm using a UV/Visible spectrophotometer (Kontron Instruments, Germany), and cell mass was estimated by using calibrated standard curves [ 24].
Rats were placed in a Kopf stereotaxic frame (Kopf Instruments) and VM cell suspension or vehicle (HBSS-glucose 0.2% DNAse) was injected in four deposits distributed in 2 AP tracts, using a 22-gauge, 10 µl Hamilton syringe.
The plates were placed for 15 min at room temperature to dissolve the dyes, and then the absorbance was examined at 595 nm by Ultra Microplate Reader ELX808IU (BioTek Instruments, VT, USA) and cell viability was calculated as a percentage of control cells treated without hollow gold nanoshells.
At 24, 48 and 72 h after drug-washout the number of cells was evaluated by counting the cells with a Coulter Counter instrument, and the cells were fixed in 70% ethanol and kept at 4°C before DNA staining with propidium iodide.
Nuclei were then analyzed on a FACS Scan instrument (Becton Dickinson), and cell cycle fractions were quantified with CellQuest software (Becton Dickinson).
However, GCT using fish embryos and/or hatchlings as recipients requires sophisticated instruments and skills for cell transplantation into the blastodisc of embryos or the peritoneal cavity of the small, sometimes only a few millimeters long larvae.
For this purpose, we undertook a range of experiments applying our modified IN Cell Analyzer 1000 instrument to various dye and cell samples.
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