LifeCanvas Technologies is pioneering holistic, intact-sample tissue processing with its sample-to-dataset pipeline and tools, seamlessly integrating each step in the workflow. As the only company to own the entire workflow from tissue preservation, clearing, antibody labeling, and refractive index-matching, to SPIM imaging and analysis, we are positioned to break down barriers and empower researchers with our streamlined approaches to ensure the best results and data possible.
Seamless data acquisition requires getting the first step right. Our novel SHIELD (Park et al, Nature Biotech, 2018) tissue preservation technique forms intramolecular bonds using polyfunctional, flexible epoxides to stabilize tissue architecture and safeguard the sample’s endogenous fluorescence, protein antigenicity, and nucleic acids without the variability of hydrogel embedding.
SHIELD is the best preservation technique to safeguard endogenous fluorescence, protein antigenicity, nucleic acids and overall tissue architecture. SHIELD preservation follows a simple protocol with no hydrogel embedding step required leading to a more repeatable protocol. Other techniques, such as PFA preservation, can lead to information loss due to physical or chemical stressors experienced during processing steps. SHIELD also protects tissue for multi-round processing allowing for different individual rounds of FP-imaging and antibody labeling on the same sample.
Our active clearing system employs a patent-pending stochastic electrotransport (Kim et al, PNAS, 2015) mechanism to foster rapid delivery of detergent SDS into tissues, facilitating uniform removal of light-scattering membrane lipids. Thanks to the application of a rotational electric field, the tissue is cleared without any damage or deformation while ensuring preservation of endogenous fluorescence proteins.
SmartClear II Pro is compatible with tissue samples preserved with SHIELD, SWITCH, and others. These methods avoid the loss of endogenous fluorescence and tissue shrinkage present in organic solvent-based clearing methods like BABB, iDisco, uDisco, 3DISCO, and vDISCO. Compared to passive clearing methods, SmartClear II Pro offers orders of magnitude faster clearing with an easy-to-use interface to increase clearing throughput. We have continuously improved upon the original electrophoretic clearing design, introducing stochastic electrotransport and a dual buffer-membrane system. These improvements ensure homogenous delipidation while preventing black particle formation, all with economic and user-friendly buffers that last up to 10 days.
By combining two powerful technologies – stochastic electrotransport (Kim et al, PNAS, 2015) and SWITCH (Murray et al, Cell, 2015), SmartLabel allows achieving whole-organ antibody labeling that is uniform from surface to core. Visualize proteins deep within the organ and study the fine-scale topography of the cells they identify.
Compared to passive labeling techniques, SmartLabel provides unprecedented penetration and uniformity of signal. There is no other reagent or product available that can label large, whole organ samples in a matter of 1-2 days. Passive labeling of large tissues is plagued by prohibitively long staining times (~weeks) and penetration issues caused by antibody depletion.
After clearing and optional labeling, translucent, delipidated samples are incubated in EasyIndex, raising and homogenizing their refractive index and turning them fully transparent. This increases light penetration into the sample and ensures that images will exhibit the highest resolution and focus. The cleared and labeled samples are then embedded in an agarose gel to stabilize them while imaging, guaranteeing alignment across channels and aiding colocalization of signals.
Built from the ground up, SmartSPIM is an advanced light sheet microscope capable of imaging large tissue samples with high speed and resolution. The system oﬀers superior imaging speed to two-photon and confocal while ensuring uniform axial resolution across the entire sample, generating datasets with 1.8 x 1.8 µm pixel size in XY and with 2 µm Z-steps (3.6x, 0.2 NA objective, ~4.5 µm tall PSF). With rapid 4-color acquisition (488, 561, 642, & 785 nm laser lines) of your embedded samples, we can precisely overlay multiplexed immunoﬂuorescence signals and ﬂuorescent protein expression patterns alike.
The axially swept light sheet design of the SmartSPIM provides unprecedented resolution and uniformity over a large field of view. It is optimized for imaging large, cleared tissue samples with its large imaging chamber, easy to use interface and rapid volumetric acquisition rate.
New to tissue clearing? Don’t worry – we have a versatile, simple, and reliable passive clearing kit that includes everything you need! The bundle includes all the ready-made solutions to preserve, clear, and index match your samples with ease for 3-D imaging and analysis.
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