Fig 1.
Flow cytometry side scatter ssc measurements of AgNP treated cells.
Side scatter SSC measurement of cells were made after treatment with AgNP- CIT and AgNP-bPEI doses between 1μg/ml and 30 μg/ml for 24 hours. AgNP-CIT (Fig 1A, top) and AgNP-bPEI (Fig 1B, bottom).
Fig 2.
SSC measurements of AgNP-bPEI treated cells compared to other AgNP treated cells.
Comparison of SSC of the four AgNP compounds at 10 μg/ml A, left and 1 μg/ml B, right. The AgNP-bPEI showed higher values suggesting greater accumulated than the other compounds at 1μg/ml and 10 μg/ml.
Table 1.
SSC measurements of cells treated with AgNP and AuNP.
Fig 3.
Comparison of nanoparticle SSC intensity of 4 AgNP and 1μm beads.
Side scatter comparison of 4 types of 80 nm AgNP particles measured simultaneously with 1μm micron polystyrene beads. The AgNP-bPEI particles had slightly more SSC than the other 3 types of particles. These data were derived by diluting the AgNP with a defined concentration of beads until the AgNP beads were successively diluted to 106 counts per ml.
Table 2.
Surface Plasmonic Resonance (SPR) vs dose.
Fig 4.
Microscopic observation of bPEI and citrate coated NP.
Microscopic images of cells incubated with AgNP-bPEI (Fig 4A) and AgNP-CIT (Fig 4B) for 24 hours. The nuclei were stained with DAPI and the cytoplasm was stained with cell mask orange. The AgNP are represented by the white spots in the darkfield microscopy images. es. Magnification 200x.
Fig 5.
Flow cytometry cytograms of nuclei treated with AgNP-bPEI and AgNP-CIT.
Flow cytometry comparison of DAPI stained nuclei derived from cells treated with AgNP-bPEI or AgNP-CIT at doses between 1μg/ml and 30 μg/ml. Each successive higher dose resulted in greater amount of side scatter. This was presumably due to higher uptake of AgNP as shown in the cell scatter data (Figs 1 and 2) and the lack of complete cell lysis at all doses.
Fig 6.
Comparison of nuclei SSC histograms from NP treated cells.
Histograms of nuclei derived from cells treated with the 4 types of AgNP at 10 μg/ml (Fig 6A) and 30 μg/ml (Fig 6B). All samples demonstrated an increase of scatter after AgNP incubation for 24 hours at doses between 1 and 30 μg/ml. All the AgNP showed greater SSC than the AuNP.
Table 3.
Cell cycle phases of AgNP-bPEI.
Fig 7.
Mitochondria microscopy from AgNP-PVP and AgNP-bPEI treated cells.
Comparison of the mitochondria shapes derived from cells treated with 3 μg/ml of AgNP-PVP (Fig 7A) and AgNP-bPEI (Fig 7B). The AgNP-bPEI had beaded shapes while mitochondria derived from the AgNP-PVP treated cells had a linear strand like formation of mitochondria. The cytoplasm is stained with cell mask orange (designated by red), the mitochondria transfected with GFP (designated by green) and the nuclei are stained with DAPI (designated by blue). The NP (designated by white) are accumulated throughout the cytoplasm and around the nuclei with the AgNP-bPEI samples. Magnification 600x.
Fig 8.
Microscopic comparison between different AgNP.
Microscopic comparison of mitochondria after AgNP uptake into cells treated. with AgNP-bPEI, AgNP-PVP or AgNP-CIT for 24 hours. The AgNP-bPEI showed beaded structures of mitochondria at 0.3μg/ml (Fig 8A) and 3μg/ml (Fig 8B) while the AgNP-PVP 3μg/ml, (Fig 8C) and AgNP-CIT 3μg/ml,(Fig 8D) showed an elongated formation of mitochondria (designated by green color). The NP (designated by white color) are accumulated into the lysosomes (designated by red color) with the AgNP-bPEI samples while the NP are accumulated around the nuclei (Fig 8C) or in the ER (Fig 8D) with the other AgNP coated samples. The cytoplasm was stained with cell mask orange (red) and the nuclei are stained with DAPI (blue). Magnification 600x.
Fig 9.
PARISS hyperspectral imaging of cell and exterior.
The reference spectral library A was obtained by dividing the internal white xenon spectra with the spectra derived from individual NP or regions of NPs. A region of interest (ROI) was made around in the cell and another ROI was made on the exterior of the cell (Fig 9B) to determine the amount of reference spectra shown in Fig 9A that existed in these two regions that are expressed in a histogram format (Fig 9C and 9D).
Fig 10.
Hyperspectral image comparison of 4 types of AgNP.
The top row in Fig 10 consists of scattered images from the PARISS hyperspectral system, while the middle row presents pseudo-colored images using the spectral library shown in Fig 9A. The percentage of each type of spectra is shown in the histograms displayed on the bottom row of Fig 10. The magnification of the images was 600x.