کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8422519 1545947 2014 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Pre-fixation of virulent Mycobacterium tuberculosis with glutaraldehyde preserves exquisite ultrastructure on transmission electron microscopy through cryofixation and freeze-substitution with osmium-acetone at ultralow temperature
ترجمه فارسی عنوان
پیش تصحیح میکروسکوپیک توبرکلوزیس با غلظت گلوتارالدئید حفظ فوق العاده ای از نفوذ در میکروسکوپ الکترونی انتقال از طریق غربالگری و بستن جایگزینی با اسسمیا-استون در دمای فوقانی
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
چکیده انگلیسی
Sample preparations for transmission electron microscopy of virulent Mycobacterium tuberculosis are usually performed with chemical fixation using glutaraldehyde (GA) in a biosafety area followed by post-fixation with aqueous osmium tetroxide (OT) in a conventional laboratory outside the biosafety area. Freeze-substitution with osmium-acetone (OA) at ultralow temperature (− 85 °C) has been shown to provide high quality final images and preserves cellular structures intact. However, some preparation procedures for freeze-substitution often require large fixed devices for freezing in a special laboratory. We have reported a novel freeze-substitution preparation method that can be performed using a portable device in a biosafety cabinet at biosafety level (BSL) 3 areas. Here, as a next step, we examined whether images obtained from rapid freeze-substitution (RFS) after fixation with glutaraldehyde (GA > RFS) are of comparable quality to those obtained using standard RFS. GA > RFS provided excellent preservation of mycobacterial cell ultrastructure, including visualization of cytoplasmic ribosomes, DNA fibers, and the outer membrane. The average number of ribosomes per cubic micrometer counted on RFS and GA > RFS was not significantly different (6987.8 ± 2181.0 and 6888.9 ± 1799.3, respectively). These values were higher, but not significantly so, than those obtained using conventional chemical fixation (5018.7 ± 2511.3). This procedure may be useful for RFS preparation of unculturable mycobacteria strains or virulent strains isolated in laboratories that cannot perform RFS.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 96, January 2014, Pages 50-55
نویسندگان
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