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En este artículo

  • Overview
  • Protocolo
  • Divulgaciones
  • Materiales
  • Referencias

Overview

This video demonstrates the differential staining of collagen fibers, nuclei, and cytoplasm in heart tissue to visualize fibrotic growth associated with cardiac pathologies.

Protocolo

1. Masson-Goldner Trichrome Staining

  1. Make tissue sections from paraffin-embedded heart blocks with a manual microtome (thickness: 5 µm).
  2. Stain one slide from each heart part for each rat (3 - 4 transversal sections per slide) with Masson-Goldner trichrome staining.
    1. Melt the slides at 60 °C in an oven. Under a fume hood, deparaffinize them in xylol twice for 10 min each. Rehydrate them in 100% ethanol, 95% ethanol, 70% ethanol, and distilled water for 3 min each.
    2. Fix them in Bouin solution overnight. Then, rinse them in running tap water for 10 - 15 min. Rinse them in distilled water. Incubate them in Mayer's hematoxylin for 3 min.
    3. Remove the slides and leave them in distilled water for 5 min. Then, incubate them in acid Fuchsin-Ponceau for 5 min. Rinse them in 1% acetic acid for 1 min.
    4. Next, incubate them in phosphomolybdic acid Orange G for 1 min. Rinse them in 1% acetic acid for 1 min, incubate them in light green dye for 10 min, and rinse them in 1% acetic acid for 1 min.
    5. Dehydrate them in 70% ethanol (30 sec), 95% ethanol (30 sec), and 100% ethanol (5 min). Put one drop of a resinous mounting medium on the tissue sections, cover them with coverslips, and let them dry.

Divulgaciones

No conflicts of interest declared.

Materiales

NameCompanyCatalog NumberComments
Mayer hematoxylinMHS32-1LSigma Aldrich
Acid Fuchsin CI 42685F8129-50GSigma Aldrich
Ponceau Xylidin CI 16150P2395-25GSigma Aldrich
Orange G CI 16230O3756-100GSigma Aldrich
Light green CI 42095L5382-25GSigma Aldrich
Xylol10315083HoneyWell
Ethanol absolute10303990HoneyWell
Sterelogical microscopeSM2800Nikon
Formaldehyde99340Reactolab
Embedding cassetteK113.1Carl Roth
Bersoft Image measurement SoftwareBersoft.comLicensed software

Referencias

This article has been published

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Source: Valentin, J., et al. Histological Quantification of Chronic Myocardial Infarct in RatsJ. Vis. Exp. (2016). 

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