Journal: Journal of Nanobiotechnology
Article Title: Exploiting the anticancer effects of a nitrogen bisphosphonate nanomedicine for glioblastoma multiforme
doi: 10.1186/s12951-021-00856-x
Figure Lengend Snippet: Characterisation and intracellular delivery of RALA/AF647-RIS NPs. a Mean hydrodynamic size and zeta potential of RALA/AF647-RIS NPs. Particles were formulated at mass ratio 10 with 1 μg AF647-RIS and were subsequently measured using the Malvern Zetasizer Nano ZS instrument at 20 °C through DLS and laser doppler electrophoresis, respectively. b Encapsulation efficiency assay of RALA complexed AF647-RIS NPs at a range of mass ratios. Sample fluorescence was measured at 666 nm using the Cytation™5 Cell imaging Multi-Mode reader (Biotek, USA). The fluorescence intensity of AF647-RIS only control was taken as 100% fluorescence and 0% encapsulation, therefore any detected fluorescence of samples was taken as un-encapsulated fluorescent BP. The percentage of un-encapsulated AF647-RIS was used to determine the percentage of encapsulated AF647-RIS. The intracellular uptake of AF647-RIS NPs in c LN229 and d T98G cells (white arrows denote red fluorescence of AF647-RIS). Particles were prepared over a range of mass ratios containing 0.1 μg AF647-RIS. Cells were subsequently treated with RALA/AF647-RIS NPs for 2 h before flow cytometry. Confocal microscopic images exhibit the intracellular localisation of uncomplexed and RALA complexed AF647-RIS 2 h post-treatment. Slides were imaged using a TCS SP8-Leica Microsystems confocal microscope (Leica, UK) with a 63x oil objective lens and subsequently analysed using LAS AF Lite Software (Leica, UK). Results are reported as mean ± SEM, n=3, and images are representative of three independent repeats. Scale bars = 20 μM
Article Snippet: AlexaFluor®647-Risedronate (AF647-RIS) (MW: 1198 g/mol) (BioVinc, California, USA) was reconstituted in TE buffer to give a final concentration of 1 μg/μL.
Techniques: Electrophoresis, Fluorescence, Imaging, Flow Cytometry, Microscopy, Software
