A method to fix and permeabilize isolated adult mouse cardiomyocytes for immuno-staining and confocal imaging
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
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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.
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>
Posted 11 May, 2011
A method to fix and permeabilize isolated adult mouse cardiomyocytes for immuno-staining and confocal imaging
Posted 11 May, 2011
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
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