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Tissue Optical Clearing Reagent
Whole-brain/ body imaging with single-cell resolution

CUBIC

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

1. Link

2. Benefits of CUBIC

✔ 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

3. CUBIC Applications

  Fixation Wash Preprocessing Delipidation Wash Permeabilization Clearing Imaging
Perfusion fixation + post fixation 4% PFA/PBS(-) PBS 50% Scale CUBIC-1 Solution Scale CUBIC-1 Solution Scale CUBIC-1 Solution PBS 50% Scale CUBIC-2 Solution Scale CUBIC-1 Solution Mounting**
Processing temperature 4 °C RT RT 37°C 37°C RT - 37°C RT RT - 37°C RT
Processing time 1 day 1 day 1 day 2 days 6 - 8 days 1 day 1 day 2 days  

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

 

  Brain Heart Lung Kindney Liver
PBS
CUBIC

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

 

  Brain Heart Liver
PBS
CUBIC

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

4. CUBIC Application for fluorescent protein expression

  Fixation Wash Preprocessing Delipidation Wash
Perfusion fixation + post fixation
4% PFA/PBS(-)
[pH 7.6 - 7.8]
PBS 50% Scale CUBIC-1 Solution Scale CUBIC-1 Solution Scale CUBIC-1 Solution PBS
Processing temperature 4 °C RT RT 37 °C 37 °C RT - 37°C
Processing time 1 day 1 day 1 day 2 days 6 - 8 days* 1 day
  Fluorescent labels Wash Permebilization Clearing Imaging
PI(5 μg / mL) PB(10 mM) 50 % Scale CUBIC-2
Solution
Scale CUBIC-2
Solution
Mounting:**
Processing temperature RT RT RT RT - 37 °C RT
Processing time 2 days > 1day 1 day 2 days  

 

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

5. CUBICApplication 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
HE stain
Human
Lymph node
CUBIC
HE stain

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

 

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

6. 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).

7. Product list

Product Name Grade Pkg. Size Wako Cat. No. Storage Condition
CUBIC Trial Kit Tissue Optical Clearing Reagent 1 kit

290-80801

Keep at RT
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