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Whole-mouse clearing and imaging at the cellular level with vDISCO.
Cai, Ruiyao; Kolabas, Zeynep Ilgin; Pan, Chenchen; Mai, Hongcheng; Zhao, Shan; Kaltenecker, Doris; Voigt, Fabian F; Molbay, Muge; Ohn, Tzu-Lun; Vincke, Cécile; Todorov, Mihail I; Helmchen, Fritjof; Van Ginderachter, Jo A; Ertürk, Ali.
Afiliación
  • Cai R; Institute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Munich, Munich, Germany.
  • Kolabas ZI; Institute for Stroke and Dementia Research, Klinikum der Universität München, Ludwig Maximilian University of Munich, Munich, Germany.
  • Pan C; Institute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Munich, Munich, Germany.
  • Mai H; Institute for Stroke and Dementia Research, Klinikum der Universität München, Ludwig Maximilian University of Munich, Munich, Germany.
  • Zhao S; Graduate School of Systemic Neurosciences (GSN), Munich, Germany.
  • Kaltenecker D; Institute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Munich, Munich, Germany.
  • Voigt FF; Institute for Stroke and Dementia Research, Klinikum der Universität München, Ludwig Maximilian University of Munich, Munich, Germany.
  • Molbay M; Institute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Munich, Munich, Germany.
  • Ohn TL; Institute for Stroke and Dementia Research, Klinikum der Universität München, Ludwig Maximilian University of Munich, Munich, Germany.
  • Vincke C; Institute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Munich, Munich, Germany.
  • Todorov MI; Institute for Stroke and Dementia Research, Klinikum der Universität München, Ludwig Maximilian University of Munich, Munich, Germany.
  • Helmchen F; Institute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Munich, Munich, Germany.
  • Van Ginderachter JA; Institute for Stroke and Dementia Research, Klinikum der Universität München, Ludwig Maximilian University of Munich, Munich, Germany.
  • Ertürk A; Institute for Diabetes and Cancer, Helmholtz Munich, Munich, Germany.
Nat Protoc ; 18(4): 1197-1242, 2023 04.
Article en En | MEDLINE | ID: mdl-36697871
ABSTRACT
Homeostatic and pathological phenomena often affect multiple organs across the whole organism. Tissue clearing methods, together with recent advances in microscopy, have made holistic examinations of biological samples feasible. Here, we report the detailed protocol for nanobody(VHH)-boosted 3D imaging of solvent-cleared organs (vDISCO), a pressure-driven, nanobody-based whole-body immunolabeling and clearing method that renders whole mice transparent in 3 weeks, consistently enhancing the signal of fluorescent proteins, stabilizing them for years. This allows the reliable detection and quantification of fluorescent signal in intact rodents enabling the analysis of an entire body at cellular resolution. Here, we show the high versatility of vDISCO applied to boost the fluorescence signal of genetically expressed reporters and clear multiple dissected organs and tissues, as well as how to image processed samples using multiple fluorescence microscopy systems. The entire protocol is accessible to laboratories with limited expertise in tissue clearing. In addition to its applications in obtaining a whole-mouse neuronal projection map, detecting single-cell metastases in whole mice and identifying previously undescribed anatomical structures, we further show the visualization of the entire mouse lymphatic system, the application for virus tracing and the visualization of all pericytes in the brain. Taken together, our vDISCO pipeline allows systematic and comprehensive studies of cellular phenomena and connectivity in whole bodies.
Asunto(s)

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Encéfalo / Imagenología Tridimensional Límite: Animals Idioma: En Revista: Nat Protoc Año: 2023 Tipo del documento: Article País de afiliación: Alemania

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Encéfalo / Imagenología Tridimensional Límite: Animals Idioma: En Revista: Nat Protoc Año: 2023 Tipo del documento: Article País de afiliación: Alemania