Abstract
We present a reproducible protocol for fabrication of polyacrylamide (PAA) hydrogel-based nano-patterns and nano-textures with a wide range of elastic rigidities to study fundamental cell behaviors, such as mechanosensitivity and motility. We explore the benefits of this protocol by successfully testing the compatibility of the PAA platforms with super-resolution microscopy, which is largely unavailable with platforms of nano-scale textures made from different polymers. We also utilized soft and rigid nano-textures to study the mechanosensing basis of T cell behavior and phenotype. For complete information on the generation and use of this protocol, please refer to Tabdanov et al. (2018b).
| Original language | English (US) |
|---|---|
| Article number | 100013 |
| Journal | STAR Protocols |
| Volume | 1 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jun 19 2020 |
Bibliographical note
Funding Information:P.P.P. and this work were supported by a Research Scholar Grant, RSG-14-171-01-CSM from the American Cancer Society and the NIH (U54CA210190 University of Minnesota Physical Sciences in Oncology Center Project 2 to P.P.P. R01CA181385 to P.P.P. and R01CA181385S1 to E.D.T. and P.P.P.), UMN College of Science and Engineering (P.P.P.), Masonic Cancer Center (P.P.P.), and grants from the UMN Institute for Engineering in Medicine (P.P.P.), and The Randy Shaver Research and Community Fund (P.P.P.). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the NIH or other funding agencies. The University Imaging Center (http://uic.umn.edu) at the University of Minnesota facilitates use of the Nikon A1Rsi Confocal microscope and its integral 2D/3D image analysis software (NIS-Elements Confocal, NIS-Elements Confocal Advanced Research). STED microscopy was performed in the NHLBI light microscopy core facility with assistance from Drs. Christian A. Combs and Daniela Malide, supported by the NHLBI intramural research program. We thank the University Imaging Center (http://uic.umn.edu) at the University of Minnesota, UIC staff, and in particular Dr. Guillermo Marqués, for helpful assistance. Portions of this work were conducted in the Minnesota Nano Center, which is supported by the National Science Foundation through the National Nano Coordinated Infrastructure Network (NNCI) under Award Number ECCS-1542202. We thank the members of the Provenzano laboratory for insightful comments regarding this work. E.D.T. participated in the design, execution, and analysis of all nano-patterning and experiments. E.D.T. developed nano-patterning and nano-texture molding methods, and all platforms employed in this study. A.S.Z. participated in the experimental design, conducted experiments, data analysis, and interpretation. V.P. performed experiments and analysis. P.P.P. participated in experimental design, data analysis, data interpretation, and secured funding. E.D.T. A.S.Z. and P.P.P. wrote the manuscript. All authors read and contributed comments to the final manuscript. E.D.T. and P.P.P. oversaw all aspects of the study. The authors declare no competing interests.
Funding Information:
P.P.P. and this work were supported by a Research Scholar Grant , RSG-14-171-01-CSM from the American Cancer Society and the NIH ( U54CA210190 University of Minnesota Physical Sciences in Oncology Center Project 2 to P.P.P., R01CA181385 to P.P.P., and R01CA181385S1 to E.D.T. and P.P.P.), UMN College of Science and Engineering (P.P.P.), Masonic Cancer Center (P.P.P.), and grants from the UMN Institute for Engineering in Medicine (P.P.P.), and The Randy Shaver Research and Community Fund (P.P.P.). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the NIH or other funding agencies. The University Imaging Center ( http://uic.umn.edu ) at the University of Minnesota facilitates use of the Nikon A1Rsi Confocal microscope and its integral 2D/3D image analysis software (NIS-Elements Confocal, NIS-Elements Confocal Advanced Research). STED microscopy was performed in the NHLBI light microscopy core facility with assistance from Drs. Christian A. Combs and Daniela Malide, supported by the NHLBI intramural research program. We thank the University Imaging Center ( http://uic.umn.edu ) at the University of Minnesota, UIC staff, and in particular Dr. Guillermo Marqués, for helpful assistance. Portions of this work were conducted in the Minnesota Nano Center, which is supported by the National Science Foundation through the National Nano Coordinated Infrastructure Network (NNCI) under Award Number ECCS-1542202. We thank the members of the Provenzano laboratory for insightful comments regarding this work.
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© 2020