MitoSOX (Molecular Probes, M36008), MitoTracker Red CMXRos (Cell Signaling Technology, 9082), LysoTracker Red (Invitrogen, L7526), TMRM (Biotium, 70,017) and Hoechst 33,342 (Sigma, B2261) staining were described previously [30 (link),31 (link),59 ,62–64 (link)]. Insulin- (Sigma, I0516) stimulated glucose uptake assay was measured as previously described using the fluorescent D-glucose analog 2NBDG (Invitrogen, N13195) [29 (link)]. Microscopy was performed on an Olympus IX70 inverted microscope (Toronto, ON, Canada) with QImaging Retiga SRV Fast 1394 camera (Surrey, BC, Canada) using NIS Elements AR 3.0 software (Nikon Instruments Inc., Melville, NY, USA), or a Zeiss Axiovert 200 inverted microscope fitted with a Calibri 7 LED Light Source and Axiocam 702 mono camera (Pleasanton, USA) [30 (link),59 ]. Quantification, scale bars, background subtract, and processing were done on Fiji (ImageJ) and Zen 2.3 Pro software.
Protocol detail
Mitochondrial and Glucose Uptake Imaging
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : University of Manitoba
Other organizations : Athabasca University, Unité de Nutrition Humaine, Agriculture and Agri-Food Canada
Top 5 similar protocols
Variable analysis
independent variables
- Insulin stimulation
- Glucose uptake
- Mitochondrial superoxide production (MitoSOX staining)
- Mitochondrial membrane potential (MitoTracker Red CMXRos staining)
- Lysosomal/acidic organelle staining (LysoTracker Red)
- Mitochondrial membrane potential (TMRM staining)
- Nuclear staining (Hoechst 33,342)
- Cell culture conditions
- Microscopy settings
- No positive controls explicitly mentioned
- No negative controls explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!