In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue
Abstract
:1. Introduction
2. Materials and Methods
2.1. Mouse Strains and Allele Deletion
2.2. Carbon Tetrachloride (CCl4) Administration
2.3. Ischemia-Reperfusion Injury
2.4. Thymus Atrophy and Toxic Injury
2.5. Plasmids
2.6. Hydrodynamic Tail Vein Injection (HDTVI)
2.7. Trypan Blue Levels in Serum and Toxicity Analysis
2.8. Pulse and Chase
2.9. Liver Sample Collection
2.10. Immunofluorescence
2.11. iDISCO+ Clearing of Mouse Liver
2.12. Light Sheet Fluorescence Microscopy
2.13. 3D Image Reconstruction and Analysis
3. Results
3.1. TBlue Marks Dead Cells during Liver Injury and Regeneration
3.2. TBlue Detects Apoptotic Cell Death
3.3. In Vivo “Pulse and Chase” Tracking of Dead Cells
3.4. In Vivo “Pulse and Chase” of Two Populations of Dead Cells
3.5. Three-Dimensional Mapping of Large Necrotic Areas in Whole Livers
3.6. Detection of Cell Death in Liver Cancer Using TBlue Labelling
3.7. Detection of Cell Death in Other Organs Using TBlue Labeling
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Acknowledgments
Conflicts of Interest
References
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Procel, N.; Camacho, K.; Verboven, E.; Baroja, I.; Guerrero, P.A.; Hillen, H.; Estrella-García, C.; Vizcaíno-Rodríguez, N.; Sansores-Garcia, L.; Santamaría-Naranjo, A.; et al. In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue. Cells 2024, 13, 1379. https://doi.org/10.3390/cells13161379
Procel N, Camacho K, Verboven E, Baroja I, Guerrero PA, Hillen H, Estrella-García C, Vizcaíno-Rodríguez N, Sansores-Garcia L, Santamaría-Naranjo A, et al. In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue. Cells. 2024; 13(16):1379. https://doi.org/10.3390/cells13161379
Chicago/Turabian StyleProcel, Nicole, Karen Camacho, Elisabeth Verboven, Isabel Baroja, Priscila A. Guerrero, Hanne Hillen, Carlos Estrella-García, Nicole Vizcaíno-Rodríguez, Leticia Sansores-Garcia, Ana Santamaría-Naranjo, and et al. 2024. "In Vivo Tracking and 3D Mapping of Cell Death in Regeneration and Cancer Using Trypan Blue" Cells 13, no. 16: 1379. https://doi.org/10.3390/cells13161379