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Method Article
Sathya Srinivasan
Experimental Imaging Centre, University of Calgary
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
We propose a simple method to fix, attach and premeabilize isolated mouse cardiomyocytes for immuno-staining and confocal microscopic imaging. This method is highly reproducible and retains the morphology of the cardiomyocytes. This method will be useful to study morphological changes in the cytoskeletal protein architecture, organellar and protein distribution within the cell without any distortion of intracellular organization. This protocol can be completed in less than 2 days.
Keywords
cardiomyocytes, confocal imaging, immunocytochemistry, method
Figure 1
The authors declare no competing financial interests
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.png
Supplementary Figure 1 Snap-shot of deconvolution confocal images of fixed isolated mouse cardiomyocytes showing different channels. Snap-shot of deconvolution confocal images of fixed isolated mouse cardiomyocytes showing different channels. (a-d) Deconvolution images obtained from a confocal microscopy with pseudo-colors. (a) nucleus stained with SYTO 11 Green-Fluorescent Nucleic Acid stain shown as green (b) F-actin filaments stained with Alexa Fluor 568 phalloidin shown as red (c) mitochondria stained with MitoTracker Deep Red 633 shown as blue, and (d) Overlay of the deconvolution image from all the channels. Scale bar, 10 μm. All experiments using animals were carried out under institutional and national guidelines.
- supplement0.mov
Supplementary Movie 1 3D cardiomyocyte Three dimentional reconstruction of isolated mouse cardiomyocyte imaged using confocal microscope. The confocal image stacks of each channels were reconstructed using Voxx (http://www.nephrology.iupui.edu/imaging/voxx/). All experiments using animals were carried out under institutional and national guidelines. <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/6heglp2OcoE&hl=en&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/6heglp2OcoE&hl=en&fs=1" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object>
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Version History
CURRENT STATUS: Posted
Version 1
Posted 11 May, 2011
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Subject Areas
Biological techniques
Associated Publications
CRYAB and HSPB2 deficiency increases myocyte mitochondrial permeability transition and mitochondrial calcium uptake
by Toshie Kadono, Xiu Quan Zhang, Sathya Srinivasan, +3
Journal Of Molecular And Cellular Cardiology (10 May, 2011)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Sathya Srinivasan
Experimental Imaging Centre, University of Calgary
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 11 May, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biological techniques
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
We propose a simple method to fix, attach and premeabilize isolated mouse cardiomyocytes for immuno-staining and confocal microscopic imaging. This method is highly reproducible and retains the morphology of the cardiomyocytes. This method will be useful to study morphological changes in the cytoskeletal protein architecture, organellar and protein distribution within the cell without any distortion of intracellular organization. This protocol can be completed in less than 2 days.
Figure 1
The authors declare no competing financial interests
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.png
Supplementary Figure 1 Snap-shot of deconvolution confocal images of fixed isolated mouse cardiomyocytes showing different channels. Snap-shot of deconvolution confocal images of fixed isolated mouse cardiomyocytes showing different channels. (a-d) Deconvolution images obtained from a confocal microscopy with pseudo-colors. (a) nucleus stained with SYTO 11 Green-Fluorescent Nucleic Acid stain shown as green (b) F-actin filaments stained with Alexa Fluor 568 phalloidin shown as red (c) mitochondria stained with MitoTracker Deep Red 633 shown as blue, and (d) Overlay of the deconvolution image from all the channels. Scale bar, 10 μm. All experiments using animals were carried out under institutional and national guidelines.
- supplement0.mov
Supplementary Movie 1 3D cardiomyocyte Three dimentional reconstruction of isolated mouse cardiomyocyte imaged using confocal microscope. The confocal image stacks of each channels were reconstructed using Voxx (http://www.nephrology.iupui.edu/imaging/voxx/). All experiments using animals were carried out under institutional and national guidelines. <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/6heglp2OcoE&hl=en&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/6heglp2OcoE&hl=en&fs=1" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object>
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Associated Publications
CRYAB and HSPB2 deficiency increases myocyte mitochondrial permeability transition and mitochondrial calcium uptake
by Toshie Kadono, Xiu Quan Zhang, Sathya Srinivasan, +3
Journal Of Molecular And Cellular Cardiology (10 May, 2011)