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. Author manuscript; available in PMC: 2013 Dec 24.
Published in final edited form as: Methods Cell Biol. 2012;111:10.1016/B978-0-12-416026-2.00015-7. doi: 10.1016/B978-0-12-416026-2.00015-7

Fig. 6.

Fig. 6

Gold nanoparticles are used as fiducial markers to align fluorescence and electron micrographs. Image is a cross-section of a C. elegans adult expressing TOM-20-tdEos. (A) A low magnification SEM image is used to orient and place the high magnification SEM image. The low magnification image (5,000×) is opened in photoshop first. A higher magnification electron micrograph (50,000×) is aligned on top of the low magnification image. High magnification image is outlined with a white line. Black dots, indicated by arrows, are the fiducial marks from 100 nm gold nanoparticles applied prior to PALM imaging. (B) A sum TIRF image is aligned onto an electron micrograph based on fluorescence from the gold fiducials. A ‘sum TIRF’ image represents all of the fluorescence obtained during the length of the imaging session. (C) A PALM image is then rotated and translated based on the values obtained in (B).