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CytoVista Blocking Buffer is used to block non-specific signal while performing immunohistochemistry (IHC) or immuno-labeling with tissue or 3D cell cultures. This buffer consists of PBS with 0.2% Triton X-100, 6% donkey serum, and 10% DMSO. It helps enable sharp and bright images at any depth of tissue or 3D cell culture when used with other CytoVista products.
Features of the CytoVista family of tissue and 3D cell culture clearing reagents include:
• Rapid clearing of fluorescently labeled tissue for 3D imaging
• Minimal shrinkage, expansion, or other morphological changes to tissue
• Simple workflow does not require any special instruments or equipment and is easily adapted to IHC
• Compatible with most fluorophores, including dyes and fluorescent proteins
The CytoVista tissue and 3D cell culture clearing workflow clears tissue or 3D cultured cells with minimal changes to morphology or the sensitivity of detection with almost any fluorophore. With an easy-to-use protocol, whole mouse brain (8-mm section) can be cleared in 24 hours, or a 1-mm section in 2 hours, without the use of any special instruments or equipment.
The CytoVista clearing workflow is compatible with most fluorophores, including fluorescent proteins that are detected with common fluorescent imaging instruments, such as wide-field microscopes, high content analyzers, confocal microscopes, and lightsheet microscopes. For first time users, we recommend the CytoVista Tissue Clearing/Staining Kit (Cat No V11324) or CytoVista 3D Cell Culture Clearing/Staining Kit (Cat No. V11325). These kits contain all necessary reagents for both clearing and IHC staining.
CytoVista clearing technology sufficiently clears tissue for 3D fluorescent imaging, but unlike other clearing techniques does not significantly changes the tissue morphology. Minimal morphological changes, such as shrinkage and contraction, have been observed. For precious samples, the clearing can be reversed, and the tissue can be processed for histology studies, like H&E staining.
For tissues that are easier to clear, like brain, lung, intestine, skin, and tissues of less than 250-µm thickness, treatment with CytoVista Tissue Clearing Reagent is sufficient for 3D imaging. However, tissues of more than 250-µm thickness or that are hard to clear like kidney, liver, heart, and placenta will require an initial treatment with CytoVista Tissue Clearing Reagent followed by treatment with CytoVista Tissue Clearing Enhancer.
CytoVista 3D Cell Culture Clearing Reagent is recommended for spheroids or organoids up to 1 mm in thickness.
For hard tissues like bone, cartilage, and connective tissue, CytoVista Tissue Clearing Reagent is not recommended.