[Histonet] H and E vs. Masson Trichrome in cardiac injury

From:"Lapointe, Nathalie,Ph.D."

> Hi,
> I found that website by hazard and I'm very impress!
> I have another questions for you:
> I'm currently exploring some hormones that may cause myocardial injury in
> adrenalectomized rats. 
> The heart were stained with hematoxylin and eosin for light microscopic
> analysis (x40). A scale from 0-4 was used to score the level of myocardial
> injury. A score of 0 represented no damage. A score of 1 represented the
> presence of myocytes demonstrating early necrotic changes such as nuclear
> pyknosis or karylysis, and oesinophil staining of the cytoplasm associated
> with the presence of scattered neutrophilic infiltrates.  A score of 2 was
> given when one clear area of necrosis (loss of myocardial cells with heavy
> neutrophilic infiltrates) was observed.  When two or more separate areas of
> necrosis were found (implicating the presence of two different myocardial
> infarctions in the same heart), but the areas localized and compromised less
> then 50% of the ventricular wall, the hearts received a score of 3. A score of
> 4 was assigned to hearts that demonstrated extensive areas of necrosis
> compromising more than 50% of wither the left or the right ventricle. 
> I'm looking for another way then the above score to measure cardiac injury.
> Based to the fact that Hematoxylin is a dark purplish dye that will stain the
> chromatin within the nucleus (therefore also monocyte infiltration), and that
> Eosin is an orangish pink to red dye that stains the cytoplasmic material
> including connective tissue and collagen (therefore doesn't stain necrotic
> tissues), I was wondering if a software in which I could measure the contrast
> of color related to injury vs. intact tissue exist? Does this same software
> (if available) can also calculated the number of dark purplish dye (therefore
> level of inflammation)?
> We also stained the same heart with Masson Trichrome. Do you think that
> measuring the % of blue staining (collagen) in the whole heart at a
> magnification of X4 is equivalent to H and E score described above? How can I
> know if the blue staining with Masson Trichrome is fibrosis rather then
> collagen or only red Sirus staining can measure fibrosis? 
Thank you very much for all your help.


> Nathalie Lapointe
> Brigham and Women's Hospital
> LMRC #214
> 221 Longwood Ave
> Boston, MA, USA
> 02115
> (617) 515-5936

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