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High-resolution 3D images of organelles are of paramount importance in cellular biology. Although light microscopy and transmission electron microscopy (TEM) have provided the standard for imaging cellular structures, they cannot provide 3D images. However, recent technological advances such as serial block-face scanning electron microscopy (SBF-SEM) and focused ion beam scanning electron microscopy (FIB-SEM) provide the tools to create 3D images for the ultrastructural analysis of organelles. Here, we describe a standardized protocol using the visualization software, Amira, to quantify organelle morphologies in 3D, thereby providing accurate and reproducible measurements of these cellular substructures. We demonstrate applications of SBF-SEM and Amira to quantify mitochondria and endoplasmic reticulum (ER) structures.
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This content will become publicly available on November 26, 2025
Bridging high resolution sub-cellular imaging with physiologically relevant engineered tissues
While high-resolution microscopic techniques are crucial for studying cellular structures in cell biology, obtaining such images from thick 3D engineered tissues remains challenging. In this review, we explore advancements in fluorescence microscopy, alongside the use of various fluorescent probes and material processing techniques to address these challenges. We navigate through the diverse array of imaging options available in tissue engineering field, from wide field to super-resolution microscopy, so researchers can make more informed decisions based on the specific tissue and cellular structures of interest. Finally, we provide some recent examples of how traditional limitations on obtaining high-resolution images on sub-cellular architecture within 3D tissues have been overcome by combining imaging advancements with innovative tissue engineering approaches.
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- Award ID(s):
- 2338931
- PAR ID:
- 10571296
- Editor(s):
- Rego, Bruno
- Publisher / Repository:
- Frontiers
- Date Published:
- Journal Name:
- Frontiers in Mechanical Engineering
- Volume:
- 10
- ISSN:
- 2297-3079
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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