產(chǎn)品詳情
showed similar results (~68 nm for RL-100 and ~52 nm for PMMA-MA) but with less regular
shapes, which can be explained by the aggregation of these polymers when present in a dried
b (Fig. 1E, S10B). Thus, PMMA-MA in dried b (TEM) displayed smaller size, which could
be explained by the differences in the size of the hydration shell and scattering efficiency.
Next to the possibility of fluorescence intensity-based ratiometric analysis using common
widefield and confocal fluorescence microscopy, the nanoparticles should also display O2 -
sensitive phosphorescence lifetime changes measurable by phosphorescence lifetime imaging
microscopy (PLIM). We perbed calibration of MMIR1 in water and 10% serum (mimicking
cell environment) and found a minor effect of serum, slightly reducing of the luminescence
decay times. This can be explained by the fact that some population of the dye is still exposed
to the ''external'' microenvironment and is not fully protected, due to the nature of the
nanoprecipitation technique. Nevertheless, the effect was minor (an average of 4%) and we
concluded that nanoparticles provide good protection against interferences from serum and
other biological components (Fig. 1F). These measurements also indicated high reproducibility
of the oxygen sensing properties for different batches of the material even if the ratio of both
emitters is slightly varied. For the subsequent experiments, we decided to focus on ratiometric
semi-quantitative detection, as the most widely available and affordable microscopy readout.
shapes, which can be explained by the aggregation of these polymers when present in a dried
b (Fig. 1E, S10B). Thus, PMMA-MA in dried b (TEM) displayed smaller size, which could
be explained by the differences in the size of the hydration shell and scattering efficiency.
Next to the possibility of fluorescence intensity-based ratiometric analysis using common
widefield and confocal fluorescence microscopy, the nanoparticles should also display O2 -
sensitive phosphorescence lifetime changes measurable by phosphorescence lifetime imaging
microscopy (PLIM). We perbed calibration of MMIR1 in water and 10% serum (mimicking
cell environment) and found a minor effect of serum, slightly reducing of the luminescence
decay times. This can be explained by the fact that some population of the dye is still exposed
to the ''external'' microenvironment and is not fully protected, due to the nature of the
nanoprecipitation technique. Nevertheless, the effect was minor (an average of 4%) and we
concluded that nanoparticles provide good protection against interferences from serum and
other biological components (Fig. 1F). These measurements also indicated high reproducibility
of the oxygen sensing properties for different batches of the material even if the ratio of both
emitters is slightly varied. For the subsequent experiments, we decided to focus on ratiometric
semi-quantitative detection, as the most widely available and affordable microscopy readout.
