HeLa cells transfected with mtGFP or MEFmtGFP cells were seeded at a density of 100,000 cells/well on glass‐bottom dishes (MatTek) coated with poly‐D‐lysine (Sigma) or rat tail collagen 1 (GIBCO). Cells were then treated as indicated and images were recorded with an Olympus IX71 microscope with 60× oil objective (Olympus), a Hamamatsu C8484 camera (Hamamatsu Photonics), and HCI image software (Hamamatsu Photonics). Quantification was performed by blinding the images and then scoring cells based on the presence of primarily fragmented, tubular, or elongated mitochondria, as before (Lebeau et al, 2018 (link)). At least three different researchers scored each set of images and these scores were averaged for each individual experiment and all quantifications shown were performed for at least three independent experiments quantifying a total of > 60 cells/condition across all experiments. The data were then prepared in PRISM (GraphPad, San Diego, CA) and plotted on a stacked bar plot to show the average morphology and standard error of the mean across all experiments. Statistical comparisons were performed using a two‐way ANOVA in PRISM, comparing the relative amounts of fragmented, tubular, or elongated mitochondria across different conditions.
Hci image software
Manufactured by Hamamatsu Photonics
HCI image software is a computer program designed for the analysis and processing of digital images. It provides a range of tools and functionalities for tasks such as image enhancement, segmentation, and feature extraction. The software is primarily intended for use in research and laboratory settings, where it can assist in the quantitative analysis of microscopic or other types of images.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using hci image software
1
Quantifying Mitochondrial Morphology Changes
2
Mitochondrial Morphology Quantification
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MEF or HeLa cells transiently transfected with mtGFP or MEFmtGFP cells were seeded at a density of 100,000 cells/well on glass-bottom dishes (MatTek) coated with poly-D-lysine (Sigma) or rat tail collagen 1 (GIBCO). Cells were then treated as indicated and images were recorded with an Olympus IX71 microscope with 60x oil objective (Olympus), a Hamamatsu C8484 camera (Hamamatsu Photonics), and HCI image software (Hamamatsu Photonics). Quantification was performed by blinding the images and then scoring cells based on the presence of primarily fragmented, tubular, or elongated mitochondria, as before (Lebeau et al., 2018 (link)). At least three different researchers scored each set of images and these scores were averaged for each individual experiment and all quantifications shown were performed for at least 3 independent experiments quantifying a total of >60 cells/condition across all experiments. The data were then analyzed in PRISM (GraphPad, San Diego, CA) and plotted on a stacked bar plot to show the average morphology and standard error of the mean across all experiments. Statistical comparisons were performed using a 2-way ANOVA in PRISM, comparing the relative amounts of fragmented, tubular, or elongated mitochondria across different conditions.
3
Mitochondrial Morphology Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cells transfected with mt GFP or MEF mtGFP cells were seeded at a density of 100,000 cells/well on glassbottom dishes (MatTek) coated with poly-D-lysine (Sigma) or rat tail collagen 1 (GIBCO). Cells were then treated as indicated and images were recorded with an Olympus IX71 microscope with 60x oil objective (Olympus), a Hamamatsu C8484 camera (Hamamatsu Photonics), and HCI image software (Hamamatsu Photonics). At least 20 cells were imaged per condition for each experiment for quantification. Quantification was performed by blinding the images and then scoring cells based on the presence of primarily fragmented, tubular, or elongated mitochondria, as before 38 (link) . Three different researchers scored each set of images and these scores were averaged for each individual experiment. All quantifications shown were performed for at least 3 independent experiments, where averages in morphology quantified from each individual experiment were then combined. The data were then prepared in PRISM (GraphPad, San Diego, CA) and plotted on a stacked bar plot to show the average morphology and standard error of the mean across all experiments.
Statistical comparisons were performed using a 2-way ANOVA in PRISM, comparing the relative amounts of fragmented, tubular, or elongated mitochondria across different conditions.
Statistical comparisons were performed using a 2-way ANOVA in PRISM, comparing the relative amounts of fragmented, tubular, or elongated mitochondria across different conditions.
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