Simple Methods to Correlate
Conclusion The presence of internal or external fiducial markers is
very important for any sequential CLEM workf low. These markers can be either added to the specimen (gold fiducials, nanospheres, etc.), be part of the specimen (features within the sample, shape, etc.), or embedded within the sample holder or substrate of choice [1,4,5,7]. In the work described here, the three CLEM procedures used were sequential and, in each case, a different fiducial marker was exploited (dia- mond scribed marks, embedded grids on epoxy blocks, and plastic section shaped together with tissue morphology). Although our imaging used commercial correlative soft- ware, the workf lows described here can be replicated using any instrument since the fiducials described are visible in both imaging platforms and should help the researcher locate the ROIs. That being said, a correlative image soft- ware program is essential for bridging the differences in resolution and enabling the integration of molecular and structural information.
Acknowledgements Microscopy was performed at the Multiscale Microscopy
Core, a University Shared Resource at OHSU. Te authors acknowledge funding from OCSSB to CSL and NIH-NCI Cancer Center Support Grant (CCSG) 2P30CA069533. Te Brenden-Colson Center for Pancreatic Care additionally funded portions of this work. We would also like to acknowl- edge all the collaborators who contributed with samples and advice.
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[2] T Muller-Reichert and P Verkade, Methods Cell Biol 111 (2012) xvii–xix.
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