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A protocol for the analysis of a binary system comprising polyacrylamide hydrogel-attached sperm cells using high-vacuum scanning electron microscopy (SEM) is presented. This protocol focuses on optimizing the SEM procedure to obtain accurate and detailed imaging of the sperm cells and their interactions with the hydrogel scaffold. The methodology involves a stepwise sample preparation, including sample dehydration through a gradual exchange of ethanol/water ratios, followed by the application of a conductive metal coating. By employing this modified protocol, the traditional use of acetone dehydration, which may introduce chemical alterations to the materials, is avoided. The proposed approach enables a comprehensive evaluation of the morphology and interactions within the biological system in contact with the soft material scaffold. Furthermore, the potential application of this protocol extends to the study of other mammalian reproductive cells or cells of different origins adhered to hydrogel scaffolds. RESEARCH HIGHLIGHTS: Novel SEM protocol reveals precise imaging of sperm-hydrogel attachment in a binary system, enhancing our understanding of cell-material interactions. By optimizing SEM procedures, the protocol achieves precise imaging of sperm-hydrogel interactions using ethanol/water dehydration and a conductive metal coating. This modified approach enables a thorough assessment of morphology and interactions in the binary system,extending its potential applicability to other reproductive cells on hydrogelscaffolds.

Titel:	Optimized protocol for high-vacuum scanning electron microscopy analysis of polyacrylamide hydrogel-attached sperm cells in a binary system.
Autor/in / Beteiligte Person:	Ebel, F ; Ramírez-Reveco, A ; Strobel, P ; Wagenknecht, L ; Rodríguez, N ; Bosch, P ; Rivarola, C
Link:	Zum Volltext
Zeitschrift:	Microscopy research and technique, Jg. 87 (2024-05-01), Heft 5, S. 1122-1127
Veröffentlichung:	New York, NY : Wiley-Liss, c1992-, 2024
Medientyp:	academicJournal
ISSN:	1097-0029 (electronic)
DOI:	10.1002/jemt.24502
Schlagwort:	Animals Male Microscopy, Electron, Scanning Vacuum Hydrogels Spermatozoa Ethanol Water Mammals Dehydration Semen Acrylic Resins
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Review Language: English [Microsc Res Tech] 2024 May; Vol. 87 (5), pp. 1122-1127. <i>Date of Electronic Publication: </i>2024 Jan 23. MeSH Terms: Dehydration* ; Semen* ; Acrylic Resins* ; Animals ; Male ; Microscopy, Electron, Scanning ; Vacuum ; Hydrogels ; Spermatozoa ; Ethanol ; Water ; Mammals References: Blois, D. A., Liaudat, A. C., Capella, V., Morilla, G., Rivero, R. E., Broglia, M. F., Barbero, C. A., Rodríguez, N., Bosch, P., & Rivarola, C. R. (2021). Interaction between hyaluronic acid semi‐interpenetrated hydrogel with bull spermatozoa: Studies of sperm attachment–release and sperm quality. Advanced Materials Interfaces, 8, 2101155. ; Bordbar, S., Lotfi Bakhshaiesh, N., Khanmohammadi, M., Sayahpour, F. A., Alini, M., & Baghaban Eslaminejad, M. (2020). Production and evaluation of decellularized extracellular matrix hydrogel for cartilage regeneration derived from knee cartilage. 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(2020). Flexible dry hydrogel with lamella‐like structure engineered via dehydration in poor solvent. Chinese Chemical Society of Chemistry, 1, 533–543. Grant Information: FOVI210036 Laboratory of Cryobiology and Sperm Functionality Analysis; PICT-2020-SERIEA-03621 National Agency for the Promotion of Research, Technological Development and Innovation, Argentina; PIP11220200102431 National Research Council and Technical Research (CONICET), Argentina Contributed Indexing: Keywords: polyacrylamide hydrogel; scanning electron microscopy; stallion sperm Substance Nomenclature: 0 (polyacrylamide gels) ; 0 (Hydrogels) ; 3K9958V90M (Ethanol) ; 059QF0KO0R (Water) ; 0 (Acrylic Resins) Entry Date(s): Date Created: 20240123 Date Completed: 20240403 Latest Revision: 20240403 Update Code: 20240403

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Optimized protocol for high-vacuum scanning electron microscopy analysis of polyacrylamide hydrogel-attached sperm cells in a binary system.