Figure 1
From: Innovative particle standards and long-lived imaging for 2D and 3D dSTORM
Long-lived- fluorescence dSTORM imaging of innovative blinking LipoParticles using the in-house developed Eternity buffer. (a) Schematic representation of our innovative blinking LipoParticle used for calibration purposes. The 1 µm polymer core in blue is surrounded by lipid layers (green) in which polymer chains (black line) bearing the fluorophores (purple stars) are immobilized. (b–e) Comparison of dSTORM LipoParticle reconstruction in classical (top) and Eternity (bottom) buffers at days 0 (b,d) and 6 (c,e). The dSTORM signals are colour-coded according to their localisation precision (5 to 60 nm, inverted rainbow scale). The size of the points is also proportional to their localisation precision within a ratio of 1/10 between the smallest and the biggest points. Measurements were conducted on LipoParticles (N = 3) from the same slide kept at 4 °C and in the dark throughout the entire experiment. (f–i) Number of blinking events (f,h) and median localisation precision (g,i), extrapolated from experiments b-e, and presented as bar charts at days 0 and 6. For each time point, N = 3 replicates (±SD). (j–n) Reconstruction by dSTORM of a LipoParticle at days 0 (j), 7 (k), 14 (l), 36 (m) and 58 (n). The same colour and size code as above was applied. Measurements were conducted on LipoParticles (N = 3) from the same slide kept at 4 °C and in the dark throughout the entire experiment. (o–q) The number of blinking events (o), the median number of photons per event (p) and the median localisation precision in nm (q) extrapolated from (j–n) are represented as a function of time. For each time point, N = 3 replicates (±SD). Table 1 summarizes the conditions used to acquire and visualise images in this figure.
