{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T13:43:19Z","timestamp":1790775799431,"version":"4.1.0"},"reference-count":20,"publisher":"Georg Thieme Verlag KG","issue":"04","license":[{"start":{"date-parts":[[2019,9,23]],"date-time":"2019-09-23T00:00:00Z","timestamp":1569196800000},"content-version":"vor","delay-in-days":357,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Eur J Dent"],"published-print":{"date-parts":[[2018,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Objective: The objective of this study was to evaluate and compare the two scanning electron microscope (SEM) preparation protocols and determine the better SEM preparation technique to study stem cells on human amniotic membrane (hAM) scaffold. Materials and Methods: Formaldehyde-based protocol and glutaraldehyde-based protocol were compared to evaluate the quality of SEM images for stem cells cultured on hAM scaffold. Results: The results suggested that formaldehyde-based protocol is better than glutaraldehyde-based protocol in terms of showing clearer topography of the membrane as well as the boarders of the cells. To provide intact surface of the SEM sample and avoid possible ruptures of the hAM or the thin cell layer, it is recommended to perform the dehydration step using graded alcohol concentrations of 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90%, one time for each and twice in 100% for 10 min each. Gold sputter-coating step is not recommended as it does not improve the image quality. Conclusions: To obtain clear SEM images, it is recommended to run a preliminary study to determine the better chemicals and conditions of sample preparation even when following preexisting protocols.<\/jats:p>","DOI":"10.4103\/ejd.ejd_401_17","type":"journal-article","created":{"date-parts":[[2018,10,3]],"date-time":"2018-10-03T06:23:25Z","timestamp":1538547805000},"page":"574-578","source":"Crossref","is-referenced-by-count":57,"title":["Optimization of scanning electron microscope technique for amniotic membrane investigation: A preliminary study"],"prefix":"10.1055","volume":"12","author":[{"given":"Saaid Al","family":"Shehadat","sequence":"additional","affiliation":[{"name":"Department of Preventive and Restorative Dentistry, College of Dental Medicine, University of Sharjah, Sharjah, UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehmet Omer","family":"Gorduysus","sequence":"additional","affiliation":[{"name":"Department of Preventive and Restorative Dentistry, College of Dental Medicine, University of Sharjah, Sharjah, UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suzina Sheikh Abdul","family":"Hamid","sequence":"additional","affiliation":[{"name":"Tissue Bank Unit, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nurul Asma","family":"Abdullah","sequence":"additional","affiliation":[{"name":"Department of Biomedicine, School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdul Rani","family":"Samsudin","sequence":"additional","affiliation":[{"name":"Department of Oral and Craniofacial Health Sciences, College of Dental Medicine, University of Sharjah, Sharjah, UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Azlina","family":"Ahmad","sequence":"additional","affiliation":[{"name":"Department of Craniofacial Sciences and Oral Biology, School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"194","published-online":{"date-parts":[[2019,9,23]]},"reference":[{"key":"key-10.4103\/1305-7456.242604-1","first-page":"489","volume-title":"Scanning electron microscopic observations on deembedded biological tissue sections: Comparison of different fixatives and embedding materials","author":"Russell","year":"1985","journal-title":"Microsc Res Tech"},{"key":"key-10.4103\/1305-7456.242604-2","first-page":"8","volume-title":"Formaldehyde, formalin, paraformaldehyde and glutaraldehyde: What they are and what they do","author":"Kiernan","year":"2000","journal-title":"Microsc Today"},{"key":"key-10.4103\/1305-7456.242604-3","first-page":"177","volume-title":"Kinetic studies of formaldehyde binding in tissue","author":"Helander","year":"1994","journal-title":"Biotech Histochem"},{"key":"key-10.4103\/1305-7456.242604-4","first-page":"1281","volume-title":"Mechanism of glutaraldehyde-protein bond formation","author":"Monsan","year":"1975","journal-title":"Biochimie"},{"key":"key-10.4103\/1305-7456.242604-5","doi-asserted-by":"crossref","unstructured":"Jeffree CE, Read ND. Ambient-and low-temperature scanning electron microscopy. Electron Microsc Plant Cell 1991. p 313-414.","DOI":"10.1016\/B978-0-12-318880-9.50013-2"},{"key":"key-10.4103\/1305-7456.242604-6","first-page":"153","volume-title":"High-resolution scanning electron microscopy of immunogold-labelled cells by the use of thin plasma coating of osmium","author":"Suzuki","year":"2002","journal-title":"J Microsc"},{"key":"key-10.4103\/1305-7456.242604-7","first-page":"424","volume-title":"A new staining method (OTO) for enhancing contrast of lipid \u2013 Containing membranes and droplets in osmium tetroxide \u2013 Fixed tissue with osmiophilic thiocarbohydrazide (TCH)","author":"Seligman","year":"1966","journal-title":"J Cell Biol"},{"key":"key-10.4103\/1305-7456.242604-8","first-page":"385","volume-title":"Confocal immunofluorescence localization of collagen types I, III, IV, V and VI and their ultrastructural organization in term human fetal membranes","author":"Malak","year":"1993","journal-title":"Placenta"},{"key":"key-10.4103\/1305-7456.242604-9","first-page":"88","volume-title":"Properties of the amniotic membrane for potential use in tissue engineering","author":"Niknejad","year":"2008","journal-title":"Eur Cell Mater"},{"key":"key-10.4103\/1305-7456.242604-10","first-page":"216","volume-title":"Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction","author":"Riau","year":"2010","journal-title":"Biomaterials"},{"key":"key-10.4103\/1305-7456.242604-11","first-page":"2506","volume-title":"Cultivation of corneal epithelial cells on intact and denuded human amniotic membrane","author":"Koizumi","year":"2000","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"key-10.4103\/1305-7456.242604-12","first-page":"106","volume-title":"The successful culture and autologous transplantation of rabbit oral mucosal epithelial cells on amniotic membrane","author":"Nakamura","year":"2003","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"key-10.4103\/1305-7456.242604-13","first-page":"800","volume-title":"Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation","author":"Ishino","year":"2004","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"key-10.4103\/1305-7456.242604-14","first-page":"93","volume-title":"Sterilized, freeze-dried amniotic membrane: A useful substrate for ocular surface reconstruction","author":"Nakamura","year":"2004","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"key-10.4103\/1305-7456.242604-15","first-page":"371","volume-title":"Optimization of amniotic membrane (AM) denuding for tissue engineering","author":"Hopkinson","year":"2008","journal-title":"Tissue Eng Part C Methods"},{"key":"key-10.4103\/1305-7456.242604-16","first-page":"58","volume-title":"Chemical constitution, physical properties, and biocompatibility of experimentally manufactured Portland cement","author":"Hwang","year":"2011","journal-title":"J Endod"},{"key":"key-10.4103\/1305-7456.242604-17","first-page":"3319","volume-title":"How to optimize seeding and culturing of human osteoblast-like cells on various biomaterials","author":"Wiedmann-Al-Ahmad","year":"2002","journal-title":"Biomaterials"},{"key":"key-10.4103\/1305-7456.242604-18","first-page":"123","volume-title":"Comparison of intact and denuded amniotic membrane as a substrate for cell-suspension culture of human limbal epithelial cells","author":"Koizumi","year":"2007","journal-title":"Graefes Arch Clin Exp Ophthalmol"},{"key":"key-10.4103\/1305-7456.242604-19","first-page":"3729","volume-title":"The use of trehalose-treated freeze-dried amniotic membrane for ocular surface reconstruction","author":"Nakamura","year":"2008","journal-title":"Biomaterials"},{"key":"key-10.4103\/1305-7456.242604-20","first-page":"582","volume-title":"Human periodontal ligament cell sheets cultured on amniotic membrane substrate","author":"Adachi","year":"2014","journal-title":"Oral Dis"}],"container-title":["European Journal of Dentistry"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.thieme-connect.de\/products\/ejournals\/pdf\/10.4103\/ejd.ejd_401_17.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2019,10,25]],"date-time":"2019-10-25T04:17:03Z","timestamp":1571977023000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.thieme-connect.de\/DOI\/DOI?10.4103\/ejd.ejd_401_17"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,10]]},"references-count":20,"journal-issue":{"issue":"04","published-online":{"date-parts":[[2019,9,23]]},"published-print":{"date-parts":[[2018,10]]}},"URL":"https:\/\/doi.org\/10.4103\/ejd.ejd_401_17","relation":{},"ISSN":["1305-7456","1305-7464"],"issn-type":[{"value":"1305-7456","type":"print"},{"value":"1305-7464","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,10]]}}}