[Histonet] Protocol for fluorescence of myofibers

[J.P.H.M. van den Wijngaard]

Dear experts of histonet,

Given my formal background (chemistry synthesis) and current experience (cardiovascular research) my question may seem either too simple of not well directed. Please forgive my inexperience in these matters. 
 
In short, I am looking for a simple method that enhances the fluorescence of the cardio myofibers. 

In our institution, we have constructed a special setup allowing for investigation of vessel morphology. This is carried out by infusing a fluorescent plastic that polymerizes into an organ and then serially slicing the specimen while after each slice a high resolution image is taken of the remaining bulk material. As such, we can create high resolution 3D images of the vasculature, e.g. of a heart or kidney. 

Recently we have extended our setup (we are using a 16mpixel cooled camera which also allows very long exposure times) and are now trying to visualize the muscle fibers of the heart. For this we use a powerled (around 400nm) and image at around 600nm which seems to generates images that show autofluorescence of either collagen or muscle (I am unsure which this may be). Given these initial promising results, I would like to visualize the muscle fibers in more detail by using a staining protocol that would allow to stain post mortem hearts. I have gathered information so far that includes the use of ALA or Bouin's solution but there may be much better protocols suitable for this problem. 
I appreciate all feedback, thank you in advance,

Jeroen

Jeroen PHM van den Wijngaard, PhD

Department of Biomedical Engineering and Physics
Academic Medical Center
Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
Tel: +31 (20) 5668796