Clear a mouse brain in 6 hours!

Fig. 1: Mouse brains are cleared easily before mounting and imaging

With the Binaree® Tissue Clearing Rapid™ system the whole clearing process from fixation to imaging takes approximately 2-3 days*, which is significantly faster than many other available tissue clearing methods.

*Note: does not include primary and secondary antibody incubation times for immunostaining.

Fig. 2: Binaree Tissue Clearing Rapid is the fastest available clearing method (ref: www.binaree.com)

Passive clearing can also be performed without the Rapid system and electrophoresis, by incubating mouse brains in the Rapid clearing solution. This process takes approximately 5-7 days to achieve tissue clearing.

Fig. 3: Clear and mount mouse brains with the simple Binaree protocol

1.  Fix the mouse brain via transcardial perfusion with 4% paraformaldehyde (PFA).
2. Incubate the sample with 4% Paraformaldehyde PFA overnight at 4 °C.
3. Incubate in 30% sucrose solution at 4 °C until the sample sinks.
4. Set tissue in the electrophoresis chamber, fill with 20 mL Binaree Tissue Clearing Rapid solution, and place the chamber into the Rapid system.
   Run for 6 to 8 hours.
5. Incubate in Binaree Tissue Clearing Rapid mounting solution at 37 °C for 24-48 hours.
   The mounting solution will give the brain a uniform refractive index of 1.52.

Fig. 4: Tissue morphology and fluorescence are best preserved with Binaree clearing

2024

Therapeutic efficacy of intracerebral hematopoietic stem cell gene therapy in an Alzheimer’s disease mouse model.
Milazzo, R., Montepeloso, A., Kumar, R., Ferro, F., Cavalca, E., Rigoni, P., … & Biffi, A.
(2024). Nature Communications, 15(1), 8024.

High-Speed Clearing and High-Resolution Staining for Analysis of Various Markers for Neurons and Vessels.
Park, J. M., Choi, S. H., Lee, E. S., Gum, S. I., Hong, S., Kim, D. S., … & Oh, J. W.
(2024). Tissue Engineering and Regenerative Medicine, 1-12.

2023

Comparison of Light-Sheet Fluorescence Microscopy and Fast-Confocal Microscopy for Three-Dimensional Imaging of Cleared Mouse Brain.
Ryu, Y., Kim, Y., Park, S. J., Kim, S. R., Kim, H. J., & Ha, C. M.
(2023). Methods and Protocols, 6(6), 108.

Second Wave, Late-Stage Neuroinflammation in Cleared Brains of Aged 5xFAD Alzheimer’s Mice Detected by Macrolaser Light Sheet Microscopy Imaging.
Lee, S. H., & Son, H. J.
(2023).  International Journal of Molecular Sciences, 24(23), 17058.

2022

A new protocol for whole-brain biodistribution analysis of AAVs by tissue clearing, light-sheet microscopy and semi-automated spatial quantification.
Lopes, M. M., Paysan, J., Rino, J., Lopes, S. M., Pereira de Almeida, L., Cortes, L., & Nobre, R. J.
(2022).  Gene Therapy, 29(12), 665-679.

Three-dimensional visualization of cerebral blood vessels and neural changes in thick ischemic rat brain slices using tissue clearing.
Lee, E. J., Hong, S. K., Choi, D. H., Gum, S. I., Hwang, M. Y., Kim, D. S., … & Lee, E. S.
(2022). Scientific Reports, 12(1), 15897.