Visualization of Aspergillus fumigatus biofilms with Scanning Electron Microscopy and Variable Pressure-Scanning Electron Microscopy: A comparison of processing techniques

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• 作者：Lydia-Marie Joubert^a ; ^{lydiaj@stanford.edu} ; Jose AG Ferreira^b ; ^c ; ¹ ; David A Stevens^b ; ^c ; Hasan Nazik^b ; ^c ; ² ; Lynette Cegelski^d
• 关键词：BSE ; Backscattered Electron ; CF ; cystic fibrosis ; CPD ; critical point drying ; ECM ; extracellular matrix ; EPS ; extracellular polymeric substances ; ET ; Everhart-Thornley ; FESEM ; Field Emission Scanning Electron Microscopy ; GA ; glutaraldehyde ; HMDS ; hexamethyldisilazane ; NMR ; Nuclear Magnetic Resonance ; OsO4 ; osmium tetroxide ; PBS ; phosphate-buffered saline ; PFA ; paraformaldehyde ; SE ; Secondary Electron ; SEM ; Scanning Electron Microscopy ; SNR ; Signal to Noise Ratio ; VP-SEM ; Variable Pressure-SEM
• 刊名：Journal of Microbiological Methods
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：132
• 期：Complete
• 页码：46-55
• 全文大小：2987 K
• 卷排序：132

文摘

Visualization of A. fumigatus biofilms is influenced by SEM processing techniques. Hyphae and ECM are sensitive to interface, reagents, fixation times and drying. Shortened primary fixation, with OsO4 staining and CPD, optimally preserves structure. Contrasting with OsO4, RuO4 and Ruthenium Red enhance resolution for VP-SEM. Ionic Liquid treatment renders biofilms hydrated and conductive for SEM analysis.