The LPS (100 ng/mL) stimulated and unstimulated RAW 264.7 cells were seeded separately in the 12-well plates containing coverslips and incubated overnight. Then, the culture medium was replaced by the fresh medium containing free rhodamine B, rhodamine B-loaded calcium phosphate NPs (CaPRB NPs), and fluorescein isothiocyanate (FITC) - rhodamine B co-loaded LCaP NPs (LFCaPRB NPs) and continue culturing for another 12 h. After that, coverslips with cells were taken off and washed with ice-cold PBS to remove free dyes. The final samples were analyzed by an inverted fluorescence microscope (Leica DMIL, Germany). The ImageJ software (National Institutes of Health, USA) was used to analyze the fluorescence in each image semi-quantitatively.
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Nanoparticle Uptake in RAW 264.7 Cells
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Other organizations : Xi'an Jiaotong University, First Affiliated Hospital of Xi'an Jiaotong University
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Variable analysis
independent variables
- LPS (100 ng/mL) stimulation
- Presence of free rhodamine B
- Presence of rhodamine B-loaded calcium phosphate NPs (CaPRB NPs)
- Presence of fluorescein isothiocyanate (FITC) - rhodamine B co-loaded LCaP NPs (LFCaPRB NPs)
- Fluorescence intensity in RAW 264.7 cells
- RAW 264.7 cell type
- Incubation duration (overnight and 12 h)
- Culture medium replacement
- Washing with ice-cold PBS to remove free dyes
- Fluorescence microscopy and ImageJ analysis
- Unstimulated RAW 264.7 cells (negative control)
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