Whole-brain / body imaging with single-cell resolution

CUBIC

00595_img01.png

Dr. Hiroki R. Ueda et al. developed a simple, efficient, and scalable brain / body clearing method and imaging / computational analysis pipeline, CUBIC (clear, unobstructed brain/body imaging cocktails and computational analysis). CUBIC offers a fluorescent-preserving, high-performance and device-free tissue clearing method based on hydrophilic Solutions (ScaleCUBIC Solutions).
It enables reproducible whole-organ and whole-body clearing and rapid 3D imaging with LSFM. CUBIC also provides processing and analysis of 3D images for extracting biological information.
Therefore, CUBIC presents a platform of whole-organ/body imaging with single-cell resolution and image informatics, which enables a wide range of users to perform experiments targeting cellular and organ layers with multiple samples..


Data provided by Dr. Etsuo A Susaki, and Dr. Hiroki R. Ueda, RIKEN

Features

  • Easy-to-use
  • No special equipment required
  • Follow up 2D H&E
  • High transparency
  • Applicable for many organs
  • Compatible with IF, FP and other fluorescent labels

Protocol

Whole organ samples

2 mm section samples

Fluorescent protein expression tissue

Whole organ samples

00595_img13.png

**Typically, 3:7 mixture of Mounting Solution 1 and 2 (RI ~1.49) can be used.

Brain Heart Lung Kindney Liver
PBS 00595_img02.png
CUBIC

Figure1.Transmittance images of mouse whole organs before and after clearing with Scale CUBIC Solutions.

2 mm section samples

00595_img14.png

**Typically, 3:7 or 2:8 mixture of Mounting Solution 1 and 2 (RI ~1.49) can be used

Brain Heart Liver
PBS 00595_img03.png
CUBIC 00595_img04.png

Figure2.Transmittance images of mouse 2mm slices before and after clearing with Scale CUBIC Solutions.

Fluorescent protein expression tissue

00595_img15.png

00595_img07.png

00595_img08.png

00595_img09.png

Figure3. LSFM imaging of CAG-EGFP Tg Mouse (8w Male) whole brain, whole heart and dissected liver.

Application

CUBIC Application with pathological methods

CUBIC Application with fish clearing

CUBIC Application with crustacean clearing

CUBIC Application with beetle clearing

Visualization of whole-brain vascular networks

CUBIC Application with pathological methods

Data by S. Nojima.et al. : Scientific Reports ,7,9269 (2017) / Adapted.

Before CUBIC

After CUBIC
-------------------------------------------------------------------------------------------
0 day 2 days 3 days 4 days 6 days 7 days
Human
lung
CUBIC 00595_img10.png
HE stain
Human
Lymph
node
CUBIC 00595_img11.png
HE stain

Figure 4.Tissue clearing of various human organs with CUBIC
Tissue clearing and 3D imaging of deparaffinizedsamples with CUBIC

00595_img12.png

Figure 5.Tissue clearing and 3D imaging of pathological specimens with CUBIC.

CUBIC Application with fish clearing

Data were provided by Kunihiko Futami, Tokyo University of Marine Science and Technology.

00595_img16R.png

Figure6. Fish clearing with CUBIC
See Reference 5) for the protocol

00595_img17.png

Figure7. Stained image of Fish organ treated by CUBIC
Organs were sliced at 5 μm and stained with Mayer's hematoxylin after CUBIC treatment.

00595_img18R.png

Figure8. Immunostaining image of zebrafish treated with CUBIC

CUBIC Application with crustacean clearing

Data were provided by Aru Konno and Shigetoshi Okazaki, Hamamatsu University School of Medicine in Japan.

00595_img19.png

Figure9. Crustacean clearing with CUBIC
See Reference 6) for the protocol

00595_img20.png

Figure10. Crustacean clearing with CUBIC (PI staining)
See Reference 6) for the protocol

CUBIC Application with beetle clearing

Data were provided by Shinya Kuroda, Tokyo University

Beetle clearing requires fixing, bleaching, and clearing steps.
See the website for Detailed protocols.

00595_img21.png

Figure11. Beetle clearing with CUBIC
LEFT: drone beetle MIDDLE: rhinoceros beetle RIGHT: stag beetle
This protocol altered from reference 7).

Visualization of whole-brain vascular networks

Data by Miyawaki, T. et al. : Nat. Commun., 11:(1) (2020)

00595_img22.png

Figure12. Tissue clearing with sodium deoxycholate (SDC)
TOP: SDS-based delipidation. The tissue was swelling.
BOTTOM: SDC-based delipidation. Tissue swelling was suppressed.
This protocol altered from reference 8).

00595_img23.png

Figure13. Mouse brain cortex clearing after vascular casting
LEFT: SDS-based delipidation. Some penetrating vessels were severed (arrowheads).
BOTTOM: SDC-based delipidation. Breakages of vessels were rarely.
This protocol altered from reference 8).

References

  1. E. A. Susaki. et al. : Cell, 157(3), 726 (2014).
  2. K. Tainaka. et al. : Cell, 159(4), 911 (2014).
  3. E. A. Susaki. et al. : Nature Protocols, 10, 1709 (2015).
  4. S. Nojima. et al. : Scientific Reports, 7, 9269 (2017).
  5. S. Ohnuma. et al.: Fish Pathology, 52(2), 96(2017).
  6. A. Konno and S. Okazaki.: Zoological Letters, 4:13(2018).
  7. M Kuroda and S Kuroda (2020). Whole-body clearing of beetles by successive treatment of hydrogen peroxide and CUBIC reagents.
  8. Miyawaki, T. et al. : Nat Commun., 27, 11(2020).

Product List

  • Open All
  • Close All

For research use or further manufacturing use only. Not for use in diagnostic procedures.

Product content may differ from the actual image due to minor specification changes etc.

If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.

+49-2131-311-271

Reception hours: Mon-Fri 9:00 - 15:00 (CET)For other hours than the above, please contact us via the inquiry form.