Comprehensive Comparison Between STEM-HAADF and TEM Bright-field Mode for Imaging Resin Embedded Biological Samples.

IF 2.9 4区工程技术 Q3 MATERIALS SCIENCE, MULTIDISCIPLINARY

Microscopy and Microanalysis Pub Date : 2025-05-09 DOI:10.1093/mam/ozaf038

Kayla Lynne Haberman, Jelena Danilovic Lukovic, Snezana Kovacevic, Xinyi Zhang, Bessie Kebaara, Joseph Taube, Debora Berti, Bernd Zechmann

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引用次数: 0

Abstract

Most investigations of resin embedded biological samples by transmission electron microscopy (TEM) have been performed in TEM bright-field mode where the electron beam transmits through the sample. Modern TEMs can also be used in scanning TEM mode (STEM) where the beam scans across the sample. The preferred detector for STEM mode is the high-angle annular dark-field (HAADF) detector. The aim of this study was to compare image quality of resin embedded biological samples such as yeast (Saccharomyces), algae (Chlorella, Haematococcus), plant leaves (Nicotiana), human cells (MCF7), and animal tissue (mouse liver and brain) between TEM bright-field and STEM-HAADF mode. Generally, images taken in STEM-HAADF mode showed better image quality in terms of contrast, brightness, and signal-to-noise ratio. Samples of sections that did not receive postcontrasting with uranyl acetate or lead citrate appeared significantly less grainy. Specifically, STEM-HAADF mode resulted in significantly better image quality of algae cells, MCF7, and liver cells that did not receive postcontrasting. Artifacts visible in TEM mode were absent in STEM-HAADF mode. Thus, we can conclude that STEM-HAADF mode has significant advantages when investigating resin embedded biological samples that have little contrast or sections that did not receive postcontrasting rendering postcontrasting of sections unnecessary.

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STEM-HAADF与TEM亮场模式在树脂包埋生物样品成像中的综合比较

通过透射电子显微镜（TEM）对树脂包埋生物样品的大多数研究都是在透射电子显微镜（TEM）的亮场模式下进行的，其中电子束通过样品传输。现代TEM也可用于扫描TEM模式（STEM），其中光束扫描整个样品。STEM模式的首选探测器是高角度环形暗场（HAADF）探测器。本研究的目的是比较树脂包埋的生物样品，如酵母（Saccharomyces）、藻类（小球藻、红球藻）、植物叶片（Nicotiana）、人类细胞（MCF7）和动物组织（小鼠肝脏和大脑）在TEM亮场和STEM-HAADF模式下的图像质量。一般来说，STEM-HAADF模式下拍摄的图像在对比度、亮度、信噪比等方面表现出更好的图像质量。未接受醋酸铀酰或柠檬酸铅后对比的切片样品显着减少颗粒状。具体而言，STEM-HAADF模式显著改善了未接受后对比的藻类细胞、MCF7和肝细胞的图像质量。TEM模式下可见的工件在STEM-HAADF模式下不存在。因此，我们可以得出结论，STEM-HAADF模式在研究树脂包埋的生物样品时具有显著的优势，这些样品的对比度很小，或者没有接受后对比的切片，因此无需进行后对比。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Microscopy and Microanalysis 工程技术-材料科学：综合

CiteScore

1.10

自引率

10.70%

发文量

1391

审稿时长

6 months

期刊介绍： Microscopy and Microanalysis publishes original research papers in the fields of microscopy, imaging, and compositional analysis. This distinguished international forum is intended for microscopists in both biology and materials science. The journal provides significant articles that describe new and existing techniques and instrumentation, as well as the applications of these to the imaging and analysis of microstructure. Microscopy and Microanalysis also includes review articles, letters to the editor, and book reviews.