Fabrication of sharp silicon arrays to wound Caenorhabditis elegans.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
27 02 2020
27 02 2020
Historique:
received:
19
09
2019
accepted:
02
01
2020
entrez:
29
2
2020
pubmed:
29
2
2020
medline:
15
12
2020
Statut:
epublish
Résumé
Understanding how animals respond to injury and how wounds heal remains a challenge. These questions can be addressed using genetically tractable animals, including the nematode Caenorhabditis elegans. Given its small size, the current methods for inflicting wounds in a controlled manner are demanding. To facilitate and accelerate the procedure, we fabricated regular arrays of pyramidal features ("pins") sharp enough to pierce the tough nematode cuticle. The pyramids were made from monocrystalline silicon wafers that were micro-structured using optical lithography and alkaline wet etching. The fabrication protocol and the geometry of the pins, determined by electron microscopy, are described in detail. We also used electron microscopy to characterize the different types of injury caused by these pins. Upon wounding, C. elegans expresses genes encoding antimicrobial peptides. A comparison of the induction of antimicrobial peptide gene expression using traditional needles and the pin arrays demonstrates the utility of this new method.
Identifiants
pubmed: 32108170
doi: 10.1038/s41598-020-60333-7
pii: 10.1038/s41598-020-60333-7
pmc: PMC7046703
doi:
Substances chimiques
Caenorhabditis elegans Proteins
0
Silicon
Z4152N8IUI
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3581Références
Pujol, N. et al. Distinct innate immune responses to infection and wounding in the C. elegans epidermis. Curr. Biol. 18, 481–9, https://doi.org/10.1016/j.cub.2008.02.079 (2008).
doi: 10.1016/j.cub.2008.02.079
pubmed: 18394898
pmcid: 2394561
Hashmi, S. et al. Genetic transformation of nematodes using arrays of micromechanical piercing structures. Bio. Techniques 19, 766–770 (1995).
Trimmer, W. et al. Injection of DNA into plant and animal tissues with micromechanical piercing structures.In Proceedings IEEE Micro Electro Mechanical Systems. 1995, 111, https://doi.org/10.1109/MEMSYS.1995.472544 (IEEE, Amsterdam, Netherlands, 1995).
Hopcroft, M. A., Nix, W. D. & Kenny, T. W. What is the Young’s modulus of silicon? J. Microelectromechanical Syst. 19, 229–238, https://doi.org/10.1109/JMEMS.2009.2039697 (2010).
doi: 10.1109/JMEMS.2009.2039697
Fruhauf, J. Shape and Functional Elements of the Bulk Silicon Microtechnique (Springer-Verlag,Berlin/Heidelberg, 2005).
McAllister, D. V., Allen, M. G. & Prausnitz, M. R. Microfabricated microneedles for gene and drug delivery. Annu. Rev. Biomed. Eng. 2, 289–313, https://doi.org/10.1146/annurev.bioeng.2.1.289 (2000).
doi: 10.1146/annurev.bioeng.2.1.289
pubmed: 11701514
Larrañeta, E., Lutton, R. E., Woolfson, A. D. & Donnelly, R. F. Microneedle arrays as transdermal and intradermal drug delivery systems: Materials science, manufacture and commercial development. Mater. Sci. Eng. R: Reports 104, 1–32, https://doi.org/10.1016/j.mser.2016.03.001 (2016).
doi: 10.1016/j.mser.2016.03.001
Reed, M. & Lye, W.-K. Microsystems for Drug and Gene Delivery. Proc. IEEE 92, 56–75, https://doi.org/10.1109/JPROC.2003.820542 (2004).
doi: 10.1109/JPROC.2003.820542
Wilke, N. & Morrissey, A. Silicon microneedle formation using modified mask designs based on convex corner undercut. J. Micromechanics Microengineering 17, 238–244, https://doi.org/10.1088/0960-1317/17/2/008ISTEX (2007).
doi: 10.1088/0960-1317/17/2/008ISTEX
Jansen, H., Gardeniers, H., de Boer, M., Elwenspoek, M. & Fluitman, J. A survey on the reactive ion etching of silicon in microtechnology. J. Micromechanics Microengineering 6, 14–28, https://doi.org/10.1088/0960-1317/6/1/002ISTEX (1996).
doi: 10.1088/0960-1317/6/1/002ISTEX
Wind, R. A. & Hines, M. A. Macroscopic etch anisotropies and microscopic reaction mechanisms: a micromachined structure for the rapid assay of etchant anisotropy. Surf. Sci. 460, 21–38, https://doi.org/10.1016/S0039-6028(00)00479-9ISTEX (2000).
doi: 10.1016/S0039-6028(00)00479-9ISTEX
Belougne, J. & Caillard, C. Method and Apparatus for Etching a Substrate (2018).WO2018127561.
Dierking, K. et al. Unusual regulation of a STAT protein by an SLC6 family transporter in C. elegans epidermal innate immunity. Cell Host and Microbe 9, 425–435, https://doi.org/10.1016/j.chom.2011.04.011 (2011).
doi: 10.1016/j.chom.2011.04.011
pubmed: 21575913
Stiernagle, T Maintenance of C. elegans, vol. https://doi.org/10.1895/wormbook.1.101.1 of WormBook (The C. elegans Research Community ed, http://www.wormbook.org 2006).
Pujol, N. et al. Anti-fungal innate immunity in C. elegans is enhanced by evolutionary diversification of antimicrobial peptides Plos Pathogens, 4, https://doi.org/10.1371/journal.ppat.1000105 (2008).
doi: 10.1371/journal.ppat.1000105
Ward, J. D. et al. Defects in the C. elegans acyl-CoA synthase, acs-3, and nuclear hormone receptor, nhr-25, cause sensitivity to distinct, but overlapping stresses Plos One, 9, https://doi.org/10.1371/journal.pone.0092552 (2014).
doi: 10.1371/journal.pone.0092552
Zhu, Z. & Lui, C Anisotropic Crystalline Etching Simulation (ACES). University of Illinois (1998).
Seidel, H., Csepregi, L., Heuberger, A. & Baumgartel, H. Anisotropic Etching of Crystalline Silicon in Alkaline Solutions. J. Electrochem. Soc. 137, 15, https://doi.org/10.1149/1.2086277 (1990).
doi: 10.1149/1.2086277
Offereins, H., Kühl, K. & Sandmaier, H. Methods for the fabrication of convex corners in anisotropic etching of (100) silicon in aqueous KOH. Sensors Actuators A: Phys. 25, 9–13, https://doi.org/10.1016/0924-4247(90)87002-ZISTEX (1990).
doi: 10.1016/0924-4247(90)87002-ZISTEX
Tang, B., Sato, K. & Gosálvez, M. A. Sharp silicon tips with different aspect ratios in wet etching/drie and surfactant-modified tmah etching Sensors and Actuators A: Physical, 188, 220–229, https://doi.org/10.1016/j.sna.2012.01.031 (2012). Selected papers from The 16th International Conference on Solid-State Sensors, Actuators and Microsystems.
doi: 10.1016/j.sna.2012.01.031
Wilke, N., Reed, M. L. & Morrissey, A. The evolution from convex corner undercut towards microneedle formation: theory and experimental verification. J. Micromechanics Microengineering 16, 808–814, https://doi.org/10.1088/0960-1317/16/4/018ISTEX (2006).
doi: 10.1088/0960-1317/16/4/018ISTEX
Loer, C. M. et al. Cuticle integrity and biogenic amine synthesis in Caenorhabditis elegans require the cofactor tetrahydrobiopterin (BH4). Genetics 200, 237–53, https://doi.org/10.1534/genetics.114.174110 (2015).
doi: 10.1534/genetics.114.174110
pubmed: 25808955
pmcid: 4423366
Partridge, F. A., Tearle, A. W., Gravato-Nobre, M. J., Schafer, W. R. & Hodgkin, J. The C. elegans glycosyltransferase bus-8 has two distinct and essential roles in epidermal morphogenesis. Dev Biol 317, 549–59, https://doi.org/10.1016/j.ydbio.2008.02.060 (2008).
doi: 10.1016/j.ydbio.2008.02.060
pubmed: 18395708
Liu, J.-H., Betzner, T. M. & Henderson, H. T. Etching of self-sharpening (338) tips in (100) silicon. J. Micromechanics Microengineering 5, 18–24, https://doi.org/10.1088/0960-1317/5/1/004ISTEX (1995).
doi: 10.1088/0960-1317/5/1/004ISTEX
Schroder, H., Obermeier, E., Horn, A. & Wachutka, G. K. M. Convex corner undercutting of 100 silicon in anisotropic KOH etching: the new step-flow model of 3-d structuring and first simulation results. J. Microelectromechanical Syst. 10, 88–97, https://doi.org/10.1109/84.911096 (2001).
doi: 10.1109/84.911096
Shikida, M. et al. A model explaining mask-corner undercut phenomena in anisotropic silicon etching: a saddle point in the etching-rate diagram Sensors and Actuators A: Physical, 97–98, 758–763, https://doi.org/10.1016/S0924-4247(02)00017-1 (2002). Selected papers from Eurosenors XV.
doi: 10.1016/S0924-4247(02)00017-1
Gosálvez, M. A., Sato, K., Foster, A. S., Nieminen, R. M. & Tanaka, H. An atomistic introduction to anisotropic etching. J. Micromechanics Microengineering 17, S1–S26, https://doi.org/10.1088/0960-1317/17/4/S01ISTEX (2007).
doi: 10.1088/0960-1317/17/4/S01ISTEX
Han, J., Lu, S., Li, Q., Li, X. & Wang, J. Anisotropic wet etching silicon tips of small opening angle in KOH solution with the additions of I2/KI. Sensors and Actuators A: Physical 152, 75–79, https://doi.org/10.1016/j.sna.2009.03.008 (2009).
doi: 10.1016/j.sna.2009.03.008
Tanaka, H., Takeda, M. & Sato, K. Si (100) and (110) etching properties in 5, 15, 30 and 48 wt%KOH aqueous solution containing Triton-X-100. Microsystem Technologies 23, 5343–5350, https://doi.org/10.1007/s00542-017-3368-y (2017).
doi: 10.1007/s00542-017-3368-y
Chisholm, A. D. & Xu, S. The Caenorhabditis elegans epidermis as a model skin. ii: differentiation and physiological roles Wiley Interdiscip Rev. Dev Biol 1, 879–902, https://doi.org/10.1002/wdev.77ISTEX (2012).
doi: 10.1002/wdev.77
Taffoni, C. et al. Microtubule plus-end dynamics link wound repair to the innate immune response. eLife 9, https://doi.org/10.7554/eLife.45047 (2020).