SCALEVIEW-S
Data provided by Dr. Hiroshi Hama, Tetsushi Hoshida and Dr. Atsushi Miyawaki, Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN Biotechnological Optics Research Team, Center for Advanced Photonics, RIKEN Cooperation with Olympus
The original recipe reported by the Miyawaki team in 2011 termed Scale was an aqueous solution based on urea that is limited because the transparency process itself can damage the structures under study.
The research team spent 5 years improving the effectiveness of the original recipe to overcome this critical challenge, and the result is ScaleS, (we called SCALEVIEW-S) a new technique with many practical applications. SCALEVIEW-S creates transparent brains with minimal tissue damage, that can handle both florescent and immunohistochemical labeling techniques, and is even effective in older animals.
The new technique creates transparent brain samples that can be stored in SCALEVIEW-S solution for more than a year without damage. Internal structures maintain their original shape and brains are firm enough to permit the micron-thick slicing necessary for more detailed analyses.
Features
- Easy-to-use
- No special equipment required
- Less damage to sample
- Compatible with IF, FP and other fluorescent labels
[Use it for...]
Mouse brain, human post-mortem brain, bone*, organoid/spheroid
*Decalcification is required for bone.
Necessaries（when using SCALEVIEW-S Trial Kit)
| Reagents | Instrument |
|---|---|
|
|
Protocols
Protocol for clearing of brain slices (thickness: 1-2mm)
It is recommended to prepare slices after fixation process.
Figure 1. Transmittance images of mouse brain before and after clearing with SCALEVIEW-S Solutions.
Figure 2. Two-photon microscope imaging of YFP-H line: Mouse whole brain
| Mouse | Thy1-YFP-H line, 20W, ♂ |
| Size | Whole |
| Microscope | Olympus FVMPE-RS |
| Objective lens | XLPLN10XSVMP (NA 0.6) |
| Laser | 960 nm (for YFP) |
| Image size | 512 x 512, 170 tiles, Z=8000 μm, Z Step16 μm |
Protocol for 3-D IHC (AbScale)
It is recommended to prepare slices after fixation.
**AbScale Solution:0.33 M Urea and 0.1 % (wt/vol) Triton X-100 in PBS(-) Solution
Iba1 (RF635: Green) Amyloid-β (Alexa Fluor 488: Red) Tomato lectin (Texas Red: Blue)
Figure 3. 3D visualization of Aβ plaques (red), microglias (green) and blood vessels (blue) from a 17-month-old AD model mouse.
| Microscope (CLSM) | Olympus FV1200 |
| Objective lens | XLPLN10XSVMP (NA 0.60) |
Protocol for 3-D Chemical Staining (ChemScale)
It is recommended to prepare slices after fixation.
Figure 4. Confocal laser scanning microscope imaging of fluorescent labeled (PI) YFP-H line mouse brain slice (2 mm thick).
| Mouse | Thy1-YFP-H line, 42W, ♂ | Microscope (CLSM) | Olympus FV3000 (Inverted) |
| Size | Coronal Slice (2 mm) | Objective lens | UPLSAPO10x2 (NA 0.40) |
| Laser | 488 nm (for YFP), 561 nm (for PI ) |
Protocol for AbScale of neurosphere
*AbScale Solution:0.33 M Urea and 0.1 % (wt/vol) Triton X-100 in PBS(-) Solution
** After clearing, embed and immobilize with 1.5% (wt/vol) agarose.
Figure5. 3D visualization of Neurosphere
| Microscope (CLSM) | Olympus FV1000 |
| Objective lens | UMPLFLN10XW (NA 0.3) |
References
- Hama,H.et al. : Nature Neuroscience 14, 1481(2011).
- Hama,H.et al. : Nature Neuroscience 18, 1518(2015).
- Hama H, et al. : Protocol Exchange (2016), doi:10.1038/protex.2016.019
- Molly E, Boutin, et al :Scientific Reports, 8, 11135 (2018).
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.