Korstanje2 project protocol

Aging study: Glomerular mesangial matrix expansion in 29 inbred strains of mice   (2013)

Korstanje R
With: Noordmans GA, Caputo CR, Huang Y, Sheehan SM, Bulthuis M, Heeringa P, Hillebrands JL, van Goor H




Korstanje2 Protocol

Project protocol - Contents

Workflow and sampling

Step
Procedure
Equipment
Data collected
1
 Mice euthanized at 6, 12, 20 months of age
-
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2 Kidneys harvested and fixed Dissection kit -
3
 Histologic staining and scoring (50 glomeruli per mouse)
Microscope
Number of affected glomeruli out of 50 examined (affected=mesangial matrix expansion)

Equipment and supplies

  • Dissection kit
  • Histological sectioning equipment

Reagents and solutions

  • Bouin fixative
  • Paraffin
  • Periodic acid-Schiff staining reagent

Procedure: Kidney histologic staining and scoring glomeruli

    1. The right kidney is fixed in Bouin fixative and embedded in paraffin.
    2. Paraffin sections (3 µm) are stained with Periodic acid-Schiff reagent.
    3. Fifty glomeruli are scored for the presence or absence of mesangial matrix expansion (MME) (glomerulus is positive for MME if the mesangium is broadened more than twice).

Data collected by investigator

  • percentage of glomeruli (out of 50 examined) with mesangial matrix expansion (MME)