Protocols for Generating Surfaces and Measuring 3D Organelle Morphology Using Amira.
Algorithms
Animals
Drosophila
Endoplasmic Reticulum
GTP Phosphohydrolases
/ deficiency
Imaging, Three-Dimensional
Male
Mice, Inbred C57BL
Mice, Knockout
Microscopy, Electron, Scanning
Mitochondria
/ ultrastructure
Muscle Fibers, Skeletal
/ metabolism
Muscle, Skeletal
/ ultrastructure
Organelles
/ ultrastructure
3D imaging
3D reconstruction
Amira
FIB-SEM
SBF-SEM
mitochondrial imaging
organelles
segmentation
volume analysis
Journal
Cells
ISSN: 2073-4409
Titre abrégé: Cells
Pays: Switzerland
ID NLM: 101600052
Informations de publication
Date de publication:
27 12 2021
27 12 2021
Historique:
received:
25
09
2021
revised:
18
12
2021
accepted:
22
12
2021
entrez:
11
1
2022
pubmed:
12
1
2022
medline:
9
3
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 35011629
pii: cells11010065
doi: 10.3390/cells11010065
pmc: PMC8750564
pii:
doi:
Substances chimiques
GTP Phosphohydrolases
EC 3.6.1.-
Mfn2 protein, mouse
EC 3.6.1.-
Opa1 protein, mouse
EC 3.6.1.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NHLBI NIH HHS
ID : T32 HL007121
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK092065
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL108379
Pays : United States
Organisme : NIGMS NIH HHS
ID : R25 GM059994
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA015083
Pays : United States
Commentaires et corrections
Type : ErratumIn
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