Tissue Clearing Kit (ab243298)
Overview
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Product name
Tissue Clearing Kit
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Product overview
Tissue Clearing Kit ab243298 enables the easy set-up of rapid tissue clearing for 3D tissue imaging using simple protocols and standard laboratory equipment.
The kit enables you to set up tissue clearing easily and clear tissue quickly. The protocols are simple and use standard laboratory equipment, and can be used with immunostaining, fluorescent proteins and chemical dyes. Clearing is reversible so that you can treat your sample for conventional H&E or IHC staining after 3D imaging.
To help you get started with tissue clearing, we have tissue clearing validated antibodies. Validated antibodies are for key markers, such as NeuN, GFAP, Iba1, etc, and are tested for tissue clearing with 1 mm mouse brain sections, and also for 3D cell culture clearing with neuronal spheroids and hepatic spheroids (depending on the marker).If you want to set up tissue clearing on a sample type that we haven’t tested or to use your own antibodies, please consult the protocol book for this kit.
To learn more about tissue clearing, and tissue clearing methods (including for 3D cell cultures), please review our guide to tissue clearing.
Tissue clearing reagents and companion products are also available as individual products.
These tissue clearing reagents were developed with Visikol Inc, who developed the Visikol® HISTO™ tissue clearing technology which is the basis of our reagents.
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Notes
Simplified tissue clearing and immunostaining workflow:
- fix tissue section in your preferred fixative
- dehydrate with increasing % methanol series
- incubate with DMSO/methanol
- rehydrate with decreasing % methanol series
- incubate with Penetration Buffer
- block with Blocking Buffer
- stain with primary antibody, wash, stain with secondary antibody
- dehydrate with increasing % methanol series
- incubate with Tissue Clearing Reagent 1
- incubate with Tissue Clearing Reagent 2
- image tissue sectionThe full process, excluding fixation time and imaging, can be completed in ~ 3 days.
The workflow shown is for immuno-staining a 1 mm brain section, see kit protocol book for guidance for fluorescent proteins, labeling with chemical dyes, and other sample types / thicknesses.
Suitable sample types:
The tissue clearing reagents can be used with mammalian tissues of thicknesses ranging from conventional thin tissue sections up to whole organs.
When starting to use tissue clearing, we recommend using sections of no more than 1 mm thick; at this thickness, protocols are shorter, there are less requirements on the equipment used for imaging, and the volume of data generated is less, so analysis is easier.
We recommend consulting the kit protocol book before collecting tissue samples.
If you are preparing 1 or 2 mm mouse brain sections, we recommend our Brain Slicers.
Imaging:
Please consult the section on Imaging in the kit protocol book, before purchasing the kit. Confocal microscopes and light sheet microscopes are often suitable for 3D imaging of cleared tissues.
If you are using an air gap objective, you should also purchase a Silicon Imaging Chamber.
If you are using an immersion objective, you may require a double chambered cuvette.
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Tested applications
Suitable for: Histologymore details
Properties
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Storage instructions
Store at room temperature. Please refer to protocols. -
Components 30 ml 15 ml Tissue Clearing Antibody Buffer 1 x 30ml 1 x 15ml Tissue Clearing Blocking Buffer 1 x 30ml 1 x 15ml Tissue Clearing Penetration Buffer 1 x 30ml 1 x 15ml Tissue Clearing Permeabilization Buffer 1 x 30ml 1 x 15ml Tissue Clearing Reagent 1 1 x 30ml 1 x 15ml Tissue Clearing Reagent 2 1 x 30ml 1 x 15ml Tissue Clearing Washing Buffer 10X 1 x 70ml 1 x 35ml -
Relevance
The goal for tissue clearing is making large, fixed biological samples transparent. “Large” in this case means thick sections of tissue, whole organoids, entire organs or even entire young rats, ranging in thickness from around 100 µm to several centimeters. Typically, samples of this size are not transparent, and are therefore difficult to analyze using visible wavelengths of light with a microscope. Tissue clearing involves a series of chemical steps that render a large sample transparent.
Images
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NeuN antibody used with Tissue Clearing Kit ab243298 on 1mm mouse brain
NeuN antibody ab177487 was used with Tissue Clearing Kit ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain.
See more tissue clearing validated antibodies.
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GFAP antibody ab7260 used with Tissue Clearing Kit ab243298 on 1 mm mouse brainGFAP antibody ab7260 was used with Tissue Clearing Kit ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain.
See more tissue clearing validated antibodies.