Cells (Dec 2021)
Protocols for Generating Surfaces and Measuring 3D Organelle Morphology Using Amira
- Edgar Garza-Lopez,
- Zer Vue,
- Prasanna Katti,
- Kit Neikirk,
- Michelle Biete,
- Jacob Lam,
- Heather K. Beasley,
- Andrea G. Marshall,
- Taylor A. Rodman,
- Trace A. Christensen,
- Jeffrey L. Salisbury,
- Larry Vang,
- Margaret Mungai,
- Salma AshShareef,
- Sandra A. Murray,
- Jianqiang Shao,
- Jennifer Streeter,
- Brian Glancy,
- Renata O. Pereira,
- E. Dale Abel,
- Antentor Hinton
Affiliations
- Edgar Garza-Lopez
- Hinton and Garza Lopez Family Consulting Company, Iowa City, IA 52246, USA
- Zer Vue
- Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA
- Prasanna Katti
- National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
- Kit Neikirk
- Department of Biology, University of Hawaii at Hilo, Hilo, HI 96720, USA
- Michelle Biete
- Department of Biology, University of Hawaii at Hilo, Hilo, HI 96720, USA
- Jacob Lam
- Department of Internal Medicine, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
- Heather K. Beasley
- Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA
- Andrea G. Marshall
- Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA
- Taylor A. Rodman
- Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA
- Trace A. Christensen
- Microscopy and Cell Analysis Core Facility, Mayo Clinic, Rochester, MN 55905, USA
- Jeffrey L. Salisbury
- Microscopy and Cell Analysis Core Facility, Mayo Clinic, Rochester, MN 55905, USA
- Larry Vang
- Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN 37232, USA
- Margaret Mungai
- Department of Internal Medicine, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
- Salma AshShareef
- Department of Internal Medicine, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
- Sandra A. Murray
- Department of Cell Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 52013, USA
- Jianqiang Shao
- Central Microscopy Research Facility, University of Iowa, Iowa City, IA 52242, USA
- Jennifer Streeter
- Department of Internal Medicine, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
- Brian Glancy
- National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
- Renata O. Pereira
- Department of Internal Medicine, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
- E. Dale Abel
- Department of Internal Medicine, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
- Antentor Hinton
- Hinton and Garza Lopez Family Consulting Company, Iowa City, IA 52246, USA
- DOI
- https://doi.org/10.3390/cells11010065
- Journal volume & issue
-
Vol. 11,
no. 1
p. 65
Abstract
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.
Keywords
