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Photoconverted carbocyanine DiI allows direct visualization of transplanted glial cells at the ultrastructural level
A Gansmuller1, F Kruger, M Gumpel
1INSERM U134, Unité de Neurobiologie Cellulaire, Moléculaire et Clinique, Hôpital de la Salpêtriére, Paris, France.
Abstract:
We have used the carbocyanine fluorochrome, DiI, to trace living glial cells (astrocytes, oligodendrocytes and immortalized Schwann cells) after their transplantation into the newborn shiverer and normal mouse brain. DiI fluorescence first detected on vibratome sections, was photoconverted into a stable, non-diffusible and electron-dense diaminobenzidine product. Both fluorescence and precipitate were found in the same cells and were detectable until 60 days after transplantation. At the ultrastructural level, DiI precipitate was contained within cytoplasmic vesicles scattered in the transplanted cell bodies and processes. Photoconversion did not interfere with the cell fine structure or predicted post-transplantation behavior. DiI is thus a suitable marker to trace, at the ultrastructural level, living cells after their transplantation.