{"title":"Optimization of method for cross section hydrogels preparation using high-pressure freezing.","authors":"Shuichi Ichihashi, Masahiko Kuwata, Kodai Kikuchi, Tatsushi Matsuyama, Akio Shimizu","doi":"10.1093/jmicro/dfae020","DOIUrl":null,"url":null,"abstract":"High-pressure water freeze fracturing (HPWFF) is a method for preparing water-containing samples such as hydrogels for scanning electron microscopy, in which a sample is placed in a divisible pressure vessel, filled with water, sealed, frozen with liquid nitrogen, then vacuum dried after the vessel is divided. The pressure (about 200 MPa) generated by the phase transition from water to ice is expected to inhibit ice crystal formation that causes large deformation of microstructure in the sample. To maximize the useable sample size, where SEM observation is not affected by ice crystal growth, preparation conditions including the size of pressure vessel were examined in this work. Using pressure vessels 8.0 mm, 5.5 mm and 4.5 mm in diameter, agarose gel, gelatin gel, wheat starch hydrogel, wheat flour noodle and cellulose hydrogel were used to prepare SEM samples. With agarose gel, an area of 3.6 mm in diameter in the 5.5 mm vessel was achieved as the maximum size of the area observable without ice crystal growth. The observable size of other samples was comparable, except for gelatin gel. As a result, observation of the three-dimensional network structure of hydrogels could be performed over a wider range than with the conventional method without shredding or chemical treatment of the samples. Additionally, usability of agarose gel for sample support matrix in HPWFF was demonstrated.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.5000,"publicationDate":"2024-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microscopy","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1093/jmicro/dfae020","RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MICROSCOPY","Score":null,"Total":0}
引用次数: 0
Abstract
High-pressure water freeze fracturing (HPWFF) is a method for preparing water-containing samples such as hydrogels for scanning electron microscopy, in which a sample is placed in a divisible pressure vessel, filled with water, sealed, frozen with liquid nitrogen, then vacuum dried after the vessel is divided. The pressure (about 200 MPa) generated by the phase transition from water to ice is expected to inhibit ice crystal formation that causes large deformation of microstructure in the sample. To maximize the useable sample size, where SEM observation is not affected by ice crystal growth, preparation conditions including the size of pressure vessel were examined in this work. Using pressure vessels 8.0 mm, 5.5 mm and 4.5 mm in diameter, agarose gel, gelatin gel, wheat starch hydrogel, wheat flour noodle and cellulose hydrogel were used to prepare SEM samples. With agarose gel, an area of 3.6 mm in diameter in the 5.5 mm vessel was achieved as the maximum size of the area observable without ice crystal growth. The observable size of other samples was comparable, except for gelatin gel. As a result, observation of the three-dimensional network structure of hydrogels could be performed over a wider range than with the conventional method without shredding or chemical treatment of the samples. Additionally, usability of agarose gel for sample support matrix in HPWFF was demonstrated.
期刊介绍:
Microscopy, previously Journal of Electron Microscopy, promotes research combined with any type of microscopy techniques, applied in life and material sciences. Microscopy is the official journal of the Japanese Society of Microscopy.