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High-Speed Clearing and High-Resolution Staining for Analysis of Various Markers for Neurons and Vessels.
Park, Jung Min; Choi, Seock Hwan; Lee, Eun-Shil; Gum, Sang-Il; Hong, Sungkuk; Kim, Dong Sun; Han, Man-Hoon; Lee, Soung-Hoon; Oh, Ji Won.
Afiliación
  • Park JM; Department of Anatomy, Yonsei University College of Medicine, Seoul, Republic of Korea.
  • Choi SH; Department of Anatomy, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
  • Lee ES; BK21 Plus KNU Biomedical Convergence Program, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
  • Gum SI; Department of Urology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
  • Hong S; Bio-Medical Research Institute, Kyungpook National University, Daegu, Republic of Korea.
  • Kim DS; Institute of Biomedical Engineering Research, Kyungpook National University, Daegu, Republic of Korea.
  • Han MH; Binaree, Inc., Daegu, Republic of Korea.
  • Lee SH; Department of Anatomy, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
  • Oh JW; BK21 Plus KNU Biomedical Convergence Program, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
Tissue Eng Regen Med ; 21(7): 1037-1048, 2024 Oct.
Article en En | MEDLINE | ID: mdl-38955906
ABSTRACT

BACKGROUND:

Tissue clearing enables deep imaging in various tissues by increasing the transparency of tissues, but there were limitations of immunostaining of the large-volume tissues such as the whole brain.

METHODS:

Here, we cleared and immune-stained whole mouse brain tissues using a novel clearing technique termed high-speed clearing and high-resolution staining (HCHS). We observed neural structures within the cleared brains using both a confocal microscope and a light-sheet fluorescence microscope (LSFM). The reconstructed 3D images were analyzed using a computational reconstruction algorithm.

RESULTS:

Various neural structures were well observed in three-dimensional (3D) images of the cleared brains from Gad-green fluorescent protein (GFP) mice and Thy 1-yellow fluorescent protein (YFP) mice. The intrinsic fluorescence signals of both transgenic mice were preserved after HCHS. In addition, large-scale 3D imaging of brains, immune-stained by the HCHS method using a mild detergent-based solution, allowed for the global topological analysis of several neuronal markers such as c-Fos, neuronal nuclear protein (NeuN), Microtubule-associated protein 2 (Map2), Tuj1, glial fibrillary acidic protein (GFAP), and tyrosine hydroxylase (TH) in various anatomical regions in the whole mouse brain tissues. Finally, through comparisons with various existing tissue clearing methodologies such as CUBIC, Visikol, and 3DISCO, it was confirmed that the HCHS methodology results in relatively less tissue deformation and higher fluorescence retention.

CONCLUSION:

In conclusion, the development of 3D imaging based on novel tissue-clearing techniques (HCHS) will enable detailed spatial analysis of neural and vascular networks present within the brain.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Encéfalo / Ratones Transgénicos / Imagenología Tridimensional / Neuronas Límite: Animals Idioma: En Revista: Tissue Eng Regen Med Año: 2024 Tipo del documento: Article Pais de publicación: Corea del Sur
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Encéfalo / Ratones Transgénicos / Imagenología Tridimensional / Neuronas Límite: Animals Idioma: En Revista: Tissue Eng Regen Med Año: 2024 Tipo del documento: Article Pais de publicación: Corea del Sur