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Tissue Clearing, the X-CLARITY™ Systems and Reagents

  Overview

Tissues are inherently three dimensional in nature, which makes imaging intact tissues a necessity for a more complete study into the relationship between structure and function and the system-level study of cellular mechanisms.

Tissue clearing has become an important step for imaging tissues in 3D at single-cell resolution. The X-CLARITY™ is a collection of systems and ready-to-use reagents to standardize, simplify, and accelerate each step of the tissue clearing process.

X-CLARITY™ is based on the CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining /in situ-hybridization-compatible Tissue hYdrogel) method. With CLARITY, preserved tissues are embedded in a hydrogel matrix and lipids are actively extracted through electrophoresis to create a stable and optically transparent tissue-hydrogel hybrid that is chemically accessible for multiple rounds of antibody labeling and imaging.

A tissue sample is incubated in the X-CLARITY™ Hydrogel Solution Kit to allow hydrogel monomers to diffuse uniformly throughout the sample. The sample is then placed in the X-CLARITY™ Polymerization System. Monomers polymerize in the anaerobic environment, linking the biomolecules to a hydrogel network, which preserves molecular information and structural integrity

Once the tissue-hydrogel hybrid has been formed, the hybrid is cleared in the X-CLARITY™ Tissue Clearing System II using the ready-to-use Electrophoretic Tissue Clearing Solution. Lipids are extracted actively through electrophoresis or passively, leaving behind a stable and transparent tissue-hydrogel hybrid that is chemically accessible for molecular phenotyping

The transparent sample is then labeled with antibodies using the DeepLabel™ Antibody Staining Kit, which enhances antibody penetration into clarified tissues. Optimized for clarified tissue samples, DeepLabel™ can be used with tissues cleared with various clearing methods such as passive or active CLARITY, DISCO, 3DISCO, uDISCO, Visikol or CUBIC.

Prior to imaging, the tissue-hydrogel hybrid is placed in X-CLARITY™ Mounting Solution, a refractive index matching solution (RIMS), to homogenize the environment within the tissue to the solution. This reduces light scatter, which in turn increases optical transparency and consequently increases image quality and imaging depth.


Features of the X-CLARITY™ Systems & Reagents for Tissue Clearing:
  • A collection of systems and ready to use reagents to make electrophoretic tissue clearing simple, rapid and efficient
  • Preserved tissues are embedded in a hydrogel matrix
  • Lipids are actively extracted through electrophoresis to create a stable transparent tissue-hydrogel hybrid
  • Chemically accessible for multiple rounds of antibody labelling and imaging
  • Cytoarchitecture remains intact
  • Endogenous fluorescence proteins are preserved for fluorescence imaging
Features of the X-CLARITY™ Hydrogel Solution Kit:
  • Pre-tested for uniform and consistent tissue-hydrogel hybridization
  • Kit consists of hydrogel solution and a polymerization initiator
  • X-CLARITY™ Hydrogel Solution is acrylamide based solution to create polyacrylamide
  • Contains no bis-acrylamide or paraformaldehyde
  • X-CLARITY™ Initiator thermally releases free radicals
Features of the X-CLARITY™ Polymerization System:
  • High throughput (up to 768 samples per run
  • Fully automated vacuum, temperature and time control
  • Rapid and consistent polymerization
  • Choice of two heat blocks, one for 6×50 mL conical tubes, the other for flat-bottom multiwall plates
  • Simplifies tissue-hydrogel hybridization
  • Touch screen interface

Features of the X-CLARITY™ Tissue Clearing System II:
  • All in one solution for electrophoretic tissue clearing
  • ETC Control Tower, ETC Chamber, Buffer Reservoir, Tray, Tissue Container Holder, Tissue Containers, Electrophoretic Clearing Solution
  • Electrophoretic and passive clearing
  • Accelerates the removal of lipids while preserving the structural integrity of the sample
  • Simple, intuitive touch screen interface for setting tissue clearing conditions
  • Built in temperature control system actively cools and heats
  • Buffer is constantly circulated to maintain capacity, control temperature and eliminate clearing byproducts
  • Advanced system insures efficient, rapid and consistent tissue clearing

Features of the DeepLabel™ Antibody Staining Kit for Cleared Tissues:
  • Optimized for thick, cleared tissues
  • Increases tissue permeability and antigen-antibody binding
  • Homogenous antibody distribution
  • Site-specific labeling
  • 2.6 X greater signal-to-background
  • Vibrant imaging at subcellular resolution
  • Simple protocol with non[toxic ready to use reagents – Solution A, Solution B, Washing Buffer and X-CLARITY™ Mounting Solution
  • Compatible with virtually all antibodies and multiple clearing methods

Features of the X-CLARITY™ Mounting System:
  • Cost effective refractive index matching solution (RIMS)
  • Refractive Index (RI) is 1,460 at 25°C
  • RI stable over a wide temperature range
  • Minimizes photobleaching, preserves fluorescence signals
  • I deal for clarified and labeled tissue samples


Specifications (Click here):  Tissue Clearing, the X-CLARITY™ Systems and Reagents

Downloads:  Tissue Clearing, the X-CLARITY™ Systems and Reagents

NumberDescriptionPrice $
C1310X X-CLARITY™ Hydrogel Solution Kit, pretested hydrogel solution for uniform and consistent tissue-hydrogel hybridization, consists of Hydrogel Solution C13103 and Initiator C13104
Inquire
C13103 X-CLARITY™ Hydrogel Solution, acrylamide-based, 1L Inquire
C13104 X-CLARITY™ Polymerization Initiator, thermal free radical initiator (also known as VA-044), 2.5 g Inquire
C2001 X-CLARITY™ Polymerization System, automated, stand alone, samples can be placed in conical tubes or multi-welled plates, touch screen interface
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C20002 X-CLARITY™ Heat Block, holds 6×50 mL conical tubes, 92×134×75 mm, 1.9 kg
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C20003 X-CLARITY™ Heat Block for flat-bottom plates, 92×134×78 mm, 2.5 kg
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C30001 X-CLARITY™ Tissue Clearing System II, accelerates the removal of lipids while preserving integrity of the sample
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C13001 Electrophoretic Tissue Clearing Solution, premixed SDS based buffer optimized for use with the Tissue Clearing System II, 12×1L
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C12002 Container for tissue container C12001
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C12001 Tissue container for use with container C12002
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C12004 Mouse brain slice holder
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C12007 Whole rat brain holder
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C12010 Holder for 36 mouse brain slices, 1.5 pore Inquire
C12020 Holder for 36 mouse brain slices, 0.6 pore Inquire
C12013 Holder for 6 mouse brain slices, 1.5 pore Inquire
C12022 Holder for 6 mouse brain slices, 0.6 pore Inquire
C12011 Holder for 6 slices, 1.5 pore Inquire
C12021 Holder for 6 slices, 0.6 pore Inquire
C12012 Holder for 1 sample, 1.5 pore
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C12022 Holder for 1 sample, 0.6 pore Inquire
C12014 Holder for 48 samples, 1.5 pore Inquire
C12024 Holder for 48 samples, 0.6 pore Inquire
C12015 Holder for 192 samples, 1.5 pore Inquire
C12025 Holder for 192 samples, 0.6 pore Inquire
C33001 DeepLabel™ Antibody Staining Kit, 1×25 mL Solution A, 2×25 mL Solution B, 1×250 mL Washing Buffer
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C33002 DeepLabel™ Solution A, 25 mL Inquire
C33003 DeepLabel™ Solution B, 25 mL Inquire
C33004 DeepLabel™ Washing Buffer, 250 mL Inquire
C13101 X-CLARITY™ Mounting Solution, refractive index matching, Ri 1,460 at 25°C, stable over wide temperature range, 25 mL Inquire
C13102 X-CLARITY™ Mounting Solution, refractive index matching, Ri 1,460 at 25°C, stable over wide temperature range, 10×25 mL
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C13107 X-CLARITY™ Mounting Solution, refractive index matching, Ri 1,460 at 25°C, stable over wide temperature range, 20×25 mL Inquire