Tms f inverted phase contrast microscope

A549 cells were seeded in 6-cm dishes at a density of 5 × 10⁴ cells per dish and pre-incubated for 48 h before being treated with the indicated NK-LAAO concentrations. The cell morphology was captured at 24 h and 48 h post treatments using a Nikon TMS-F Inverted Phase Contrast Microscope (Tokyo, Japan) at 100× magnification coupled with a Dino-Eye AM423X Digital Microscope Eyepiece Camera USB 2.0 (AnMo Electronics Corporation, New Taipei City, Taiwan).

The morphologies of P5, P5PA and of P5PA-4I NPs were here investigated by optical microscopy (OM) analysis. In the performed experiments, powdery samples or aqueous dispersions were observed with a Nikon TMS-F inverted phase contrast microscope equipped with 4 $\times$ brightfield objective, 10 $\times$ phase contrast objective, LWD 20 $\times$ phase contrast objective, LWD 40 $\times$ phase contrast objective (Nikon Instruments, Inc., New York, NY, USA). Sequential images were acquired with 4 $\times ,$ 10 $\times$ , 20 $\times$ and 40 $\times$ phase contrast objectives. The video was captured with a 20 $\times$ phase contrast objective. The camera for image capture was a Moticam 10+ (10 MP) (MoticEurope S.L.U., Cabrera de Mar, Barcelona, Spain).
The aqueous dispersions of samples were prepared at P5, P5PA and P5PA-4I concentrations of 4.1, 5.3, and 5.1 mg/mL, respectively.

Twenty-four hours after seeding the cells, Aβ_1–42 was diluted in PBS to a concentration of 30 μm, aggregated with or without p-FTAA (1.5 mm stock solution) and diluted to a concentration of 30, 3, or 0.3 μm. The solution was incubated for up to 5 h at 37 °C. At the end of the aggregation period, each sample was diluted in serum-free medium to a final dilution of 1:10 (3 μm Aβ_1–42) and added to the cells. After 72 h of exposure, the cells were visualized for morphological changes and photographed using a Nikon TMS-F inverted phase contrast microscope (Nikon Instruments Inc.) equipped with an Olympus Altra20 camera using the analySIS getIT v.5 software (Olympus Soft Imaging Solutions GmbH). Furthermore, cell viability was determined using the Cell viability assay Kit II (XTT assay; Roche Diagnostics GmbH) according to the manufacturer's instructions. The absorbance at 450 nm and 750 nm was measured after 16 h using a Victor3V 1420 multilabel reader (PerkinElmer). As controls, serum-free medium with 5% (v/v) diluents or serum-free medium with 5% (v/v) diluents with 30, 3, or 0.3 μm p-FTAA were used.