Supplementary Figure 3: Single-molecule signal quality control across entire p96-well plate for 3D dSTORM acquisition. | Nature Methods

Supplementary Figure 3: Single-molecule signal quality control across entire p96-well plate for 3D dSTORM acquisition.

From: Localization-based super-resolution imaging meets high-content screening

Supplementary Figure 3

(a) Heat maps of the total number of localizations (top), mean of Gaussian fit Chi² (middle), and median of the centroid Z coordinate for all the localizations (bottom). We can notice that there is no correlation between the position inside the plate and these quantitative metadata, illustrating that this p96-well plate is suitable to achieve reproducible 3D super-resolution imaging. (b) Normalized histoplots of integrated intensity per detection (I0) grouped by well. A similar distribution can be observed from the first well (A1) until the last well (H3). (c) Comparison between extreme positions on a plate (A1 and H12) acquired automatically using HCS-SMLM with a manual acquisition done in the middle of the plate (E6). These quality control experiments are coherent with the 3D super-resolution images acquired in Figure 3, demonstrating the possibility to automatically acquire hundreds of images in 3D SMLM with the same quality compared to manual acquisitions.

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