Masson Goldner Trichrome Staining: A Technique to Visualize Collagen in Heart Sections for Cardiovascular Fibrosis Detection

Published: April 30, 2023

Abstract

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

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

Protocol

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.

Declarações

The authors have nothing to disclose.

Materials

Mayer hematoxylin MHS32-1L Sigma Aldrich
Acid Fuchsin CI 42685 F8129-50G Sigma Aldrich
Ponceau Xylidin CI 16150 P2395-25G Sigma Aldrich
Orange G CI 16230 O3756-100G Sigma Aldrich
Light green CI 42095 L5382-25G Sigma Aldrich
Xylol 10315083 HoneyWell
Ethanol absolute 10303990 HoneyWell
Sterelogical microscope SM2800 Nikon
Formaldehyde 99340 Reactolab
Embedding cassette K113.1 Carl Roth
Bersoft Image measurement Software Bersoft.com Licensed software

Tags

Play Video

Citar este artigo
Masson Goldner Trichrome Staining: A Technique to Visualize Collagen in Heart Sections for Cardiovascular Fibrosis Detection. J. Vis. Exp. (Pending Publication), e21053, doi: (2023).

View Video