Organizational transparency and 3D imaging services

Introduction to Transparency - Overview
Tissue Clearing technology refers to the use of one or more special chemical reagents (clearing agents) to treat biological tissues or organs, reducing the attenuation of light by the tissue, thereby making the biological tissue optically transparent, facilitating optical imaging observation of the tissue. Tissue clearing technology, combined with fluorescence imaging and three-dimensional reconstruction technology, allows for three-dimensional observation and analysis of the contents and structures of tissues at the cellular level, having a significant impact in the field of life sciences.

Technical Service Content

01 Tissue Clearing
After perfusion of mice, we mainly use hydrogel-based methods to clear various samples. The advantage of this method is that the tissue volume remains almost unchanged, it protects fluorescent proteins well, allows for secondary staining, and is non-toxic. The HYBRID method is a combination of the DISCO method and the CLARITY method, which better protects fluorescent proteins and improves the degree of transparency; the SHIELD method uses commercial reagents and instruments, making the clearing process faster and more efficient.

The image below shows the results after clearing the whole brain of a mouse:

02 Immunofluorescence Labeling
After tissue clearing is completed, we use our proprietary EERS (Electro-Enhanced Rapid Staining) technology to stain samples through a process of forward labeling → reverse labeling → stop incubation → forward washing → reverse washing → change washing solution → forward washing → reverse washing. For 2mm brain slices, the fastest single antibody staining takes only 4.5 hours, and the antibody consumption can be reduced by 6-10 times compared to traditional methods.

03 3D Imaging Process
Our tissue clearing technology, combined with immunofluorescence labeling, utilizes light-sheet microscopy to comprehensively obtain three-dimensional structural information of biological tissues, including nerves, blood vessels, and cells. Our clearing imaging equipment comes from Lifecanvas Technologies (LCT), specifically the MegaSPIM and SmartSPIM systems, which can obtain high-quality three-dimensional data at various scales.

The image below shows the results of GFAP astrocyte whole brain staining:

Service Features
Fast and efficient. The clearing speed is quick, and the clearing capability is strong. The clearing of a complete adult mouse brain takes only 2.5 days at most, and staining takes only 4.5 days.

Good retention effect of endogenous fluorescent proteins, while being compatible with immunostaining.

Good retention of the morphological structure of the samples.

Uses hydrogel-based reagents, which have high biological safety.

Provides solutions for tissue processing, preparation, and analysis of different sample types.

Supports three-dimensional imaging of biological tissue samples and tissue sections of various sizes.

Complete Service Process

Before the first service, you can learn about the specific technology of product clearing through the platform website. Fill out the tissue sample handover form before sending samples, detailing the specific situation of the samples. The platform will contact the project team liaison for communication, and the project team will send the samples according to the platform's requirements and provide the shipping tracking number. The platform will customize the experimental plan based on experimental needs. Progress summaries will be provided at each stage of the sample.

01 Fill out the handover form, the project team fills in the basic information and experimental requirements of the experimental samples. The platform receives the form information and contacts the project team for coordination.

02 Tissue clearing and staining, customize the experimental plan based on customer experimental needs, and the experimental process can be recorded, archived, and fed back in real-time.

03 3D imaging, imaging can be performed after tissue clearing and immunostaining are completed. The imaging time depends on the size and quantity of the samples.

04 Data processing and analysis, images can be synchronously stitched and processed during the imaging process to obtain raw data. The project team can send a portable hard drive with the samples to copy the raw data as needed.