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Tunable Substrates Discussion


We have demonstrated the use of a new support material to enhance specimen preparation techniques for EM investi- gations in the clinical sciences. Here, we described recent applications of the tunable microchip approach to dissect molecular disease states for 3D structural analysis. While this new platform is currently being implemented to directly see and reconstruct native BRCA1 assemblies formed in human breast cancer cells, there are many other areas of scientifi c inquiry that could benefi t from the use of this technology. For instance, alternative substrates also can be used to dissect active protein components in a number of other disease conditions related to infectious pathogens, cardiac defi ciencies, or neurological disorders. Broader applications for specialized microchips may include their integration into wearable technology (for example, an Apple Watch) for the detection of diagnostic or clinical biomarkers. Overall, having versatile molecular tools to determine new hallmarks of disease states also can contribute important information to improve the design of drug therapies.


Conclusion Silicon nitride microchips with surfaces modifi ed with


specifi c molecules can be used to harvest proteins from human cells. T ese “tunable” specimen supports are engineered to target a particular protein under investigation. T ese supports were used to examine viruses and protein assemblies isolated from eukaryotic cells in cryo-EM studies.


Acknowledgments T is work was supported in part by funds from the Commonwealth Health Research Board (2080914), the Concern Foundation (303872), and NIH/NCI (R01CA193578) to D.F.K. C.W. is funded through the ICTAS Doctoral Scholar’s program at Virginia Tech and the Medical Research Scholar’s program at the Virginia Tech Carilion Research Institute.


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[3] AF Brilot et al ., J Struct Biol 177 ( 2012 ) 630 – 37 . [4] D Rhinow and W Kuhlbrandt , Ultramicroscopy 108 ( 2008 ) 698 – 705 .


[5] C Yoshioka et al ., Microsc Microanal 16 ( 2010 ) 43 – 53 . [6] CJ Russo and LA Passmore , Nat Methods 11 ( 2014 ) 649 – 52 .


[7] JR Tanner et al ., J Anal Mol Tech 1 ( 1 ) (2014 ) 1 – 6 . [8] BL Gilmore et al ., Sci Rep 5 ( 2015 ) 14440 . [9] CE Winton et al ., NPJ Breast Cancer 2 ( 2016 ) 16016 . [10] R Scully et al ., Proc Natl Acad Sci USA 94 ( 1997 ) 5605 – 10 . [11] SH Scheres , J Struct Biol 180 ( 2012 ) 519 – 30 . [12] C Walmacq et al ., Mol Cell 46 ( 2012 ) 18 – 29 . [13] RS Williams et al ., Nat Struct Biol 8 ( 2001 ) 838 – 42 .


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