Figure 4.

Optical resolution measurements from wide-field (a) and laser scanning (b) modes with filter sets for imaging FITC-like fluorophores. 100 nm fluorescent beads were prepared and imaged with a 40x 0.9 NA air objective according to standard protocols (Cole, Jinadasa and Brown, 2011). Pixel data in the X and Y dimensions are shown (squares) together with Gaussian profile fits (lines). The FWHM of the fitted profiles are shown. Measured point spread functions are slightly larger than the theoretical minimum (350 nm from imaging 100 nm beads at a wavelength of 500 nm) to achieve faster imaging with reasonable FLIM photon count. Fits were generated with the MetroloJ plugin for ImageJ (http://imagejdocu.tudor.lu/doku.php?id=plugin:analysis:metroloj:start).