The ease of acquiring high-content data with the Nanoimager is supported by a range of useful analysis features. A common problem across single-molecule biology is the difficulty of interpreting complex results. To support researchers, the Nanoimager software suite contains a range of features that analyse and present data in an accessible way, as well as providing quantitative conclusions about the sample.
smFRET traces are analysed in real time, and can be viewed as individual traces or as a population average across a single acquisition or a whole series of acquisitions. Traces can be grouped into different behaviours and 2D histograms of stoichiometry versus FRET efficiency are presented for use in alternating laser excitation (ALEX) mode.
Single-particle tracking PALM data can be acquired and analysed with the Nanoimager, providing instant diffusion analysis and visualization of tracks. In super-resolution mode, the super-resolved pointillistic image is rendered in real time, with various options for filtering the localizations, enlarging particular features and selecting one or two colors at a time. Moreover, for quantitative analysis, the user can analyse the distribution of localizations along a line; can measure the time-dependency of positions, standard deviation of fitted functions and photon count for all localizations in a given area; can quantify the degree of clustering of molecules, and can measure colocalization of molecules detected in the two separate emission channels.