Abstract: FR-PO150

Elastica Masson’s Trichrome (EMT) Staining Is Useful for the Visualization of Podocyte Foot Process and Tubular Mitochondria in the Kidney

Session Information

  • Mitochondriacs and More
    November 03, 2017 | Location: Hall H, Morial Convention Center
    Abstract Time: 10:00 AM - 10:00 AM

Category: Glomerular

  • 1004 Clinical/Diagnostic Renal Pathology and Lab Medicine

Authors

  • Matsumoto, Ayumi, Osaka University Graduate School of Medicine, Suita, Japan
  • Sakaguchi, Yusuke, Osaka University Graduate School of Medicine, Suita, Japan
  • Namba, Tomoko, Osaka University Graduate School of Medicine, Suita, Japan
  • Mizui, Masayuki, Osaka University Graduate School of Medicine, Suita, Japan
  • Hamano, Takayuki, Osaka University Graduate School of Medicine, Suita, Japan
  • Isaka, Yoshitaka, Osaka University Graduate School of Medicine, Suita, Japan
  • Matsui, Isao, Osaka University Graduate School of Medicine, Suita, Japan
  • Shimada, Karin, Osaka University Graduate School of Medicine, Suita, Japan
  • Hashimoto, Nobuhiro, Osaka University Graduate School of Medicine, Suita, Japan
  • Mori, Daisuke, Osaka University Graduate School of Medicine, Suita, Japan
  • Yamaguchi, Satoshi, Osaka University Graduate School of Medicine, Suita, Japan
  • Kubota, Keiichi, Osaka University Graduate School of Medicine, Suita, Japan
  • Oka, Tatsufumi, Osaka University Graduate School of Medicine, Suita, Japan
  • Yonemoto, Sayoko, Osaka University Graduate School of Medicine, Suita, Japan
Background

Changes in microstructures of renal cells, such as foot process effacement in podocytes and mitochondrial fission in tubular cells, are tightly correlated with the development and the progression of kidney disease. Because the sizes of these structures are less than the diffraction limit of visible light, electron microscopy is usually required for their observation. Super-resolution microscopy (SRM) is also available, but the common technique for SRM requires time-consuming immunofluorescent staining for specific molecules.

Methods

To visualize the microstructures more easily, we analyzed paraffin-embedded human renal biopsy sections stained with EMT, Hematoxylin Eosin (HE), Periodic Acid-Schiff (PAS), or Periodic Acid Methenamine Silver (PAM) using structured illumination microscopy. Sections of animal kidney disease models were also analyzed.

Results

EMT-stained paraffin-embedded sections excited by 457 or 561 nm laser were useful for the visualization of podocyte foot process. We could observe foot process in the sections from minor glomerular abnormalities but not from minimal change disease. Foot process was also observable in EMT-stained normal rat kidney but not in puromycin aminonucleoside injected kidney. The other staining methods — HE, PAS, and PAM — were not applicable to the evaluation of foot process. In the tubulointerstitial area of the human kidney, EMT-stained sections excited by 561 nm laser were useful for the evaluation of mitochondria. In some patients with kidney disease, we observed short mitochondria, which suggested the progression of mitochondrial fission. We confirmed the usefulness of EMT staining for the evaluation of mitochondria by observing mitochondrial fission in endotoxin-induced kidney injury and mitochondrial swelling in ischemia reperfusion kidney injury in mice. Tubular mitochondria were also observable in HE or PAS-stained sections, but the images obtained by these staining were less clear.

Conclusion

Paraffin-embedded kidney sections stained with EMT were useful for the visualization of foot process and mitochondria in the kidney.

Funding

  • Private Foundation Support