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SHIELD 組織透明化試劑
以腦科學研究為例, 由於大腦的複雜性，傳統的腦組織連續切片已經漸漸無法滿足研究需要。在細胞分佈，連接模式，活動模式的研究等等都需要全腦成像。目前的可行方法, 都是通過澄清後的透明腦組織, 通過免疫螢光雜染後, 做層光掃描 (light sheet), 以取得全腦全貌的螢光影像.
SHIELD 的特點在於充分保護熒光蛋白（FP）和內源性分子信息. 此技術號稱四大優勢訴求：1. 快速, 2. 簡易, 3. 可信賴, 4. 安全
SHIELD 市售產品如下 :
Technical information:
250-mL SHIELD kit:
Preserve 8 whole mouse brains or samples of equivalent size. Includes:
- SHIELD Epoxy (250 mL)
- SHIELD Buffer (125 mL)
- SHIELD ON (250 mL)
500-mL SHIELD kit:
Preserve 16 whole mouse brains or samples of equivalent size. Includes:
- SHIELD Epoxy (500 mL)
- SHIELD Buffer (250 mL)
- SHIELD ON (500 mL)
Get the first step of tissue processing right with our easy & reliable solution kit
Superior Tissue Preservation
Versatile: Safeguards endogenous fluorescence, protein antigenicity, nucleic acids, and overall tissue architecture. Synergizes with a range of existing tissue processing approaches including SWITCH (Cell, 2015) and MAP (NBT, 2016). Other preservation techniques, such as PFA preservation alone, can lead to information loss due to physical or chemical stressors experienced during processing steps.
Easy, Reliable, Fast, & Safe
Easy & Reliable:
Superior tissue preservation without the variability of hydrogel embedding. Compatible with both non-fixed and PFA-fixed tissues.
Fast:
Streamlined protocol preserves samples in a matter of days with minimal steps.
Safe:
Reagents are non-toxic and can be easily disposed of after use.
Protects Tissue for Multi-Round Processing
Repeated Staining & Imaging:
Protects tissue during FP-imaging and antibody labeling across individual rounds.
About the Technology
The publications below describe the scientific advances that our technology is built upon. These methods are used in several of our whole organ tissue processing products, including SHIELD, SmartClear II Pro, and SmartLabel.
SHIELD is a tissue preservation technique that complements and enhances the standard PFA fixation of biological tissue. It requires a series of incubation steps over the course of 4-6 days to preserve a tissue sample the size of a whole mouse brain. These steps allow epoxide molecules to diffuse into the tissue and crosslink, forming a strong skeleton that anchors proteins, enzymes, nucleic acids, and other important elements of tissue architecture. This is an ideal and necessary preparation step for active clearing or immunolabeling, as exposure to harsher chemicals or environments can strip away important targets in non-SHIELD-preserved tissue. The strong fixation of native proteins also allows for multiplex labeling, making your tissue samples even more powerful and valuable!
The SWITCH method, published in Cell in 2015, introduces a simple process for controlling a wide variety of chemical reactions during tissue processing of large animal and human samples. The method ensures uniformity of preservation and immunolabeling by synchronizing chemical reactions throughout the entire sample. There are 2 basic steps to the method: SWITCH-OFF, when chemicals and buffers are allowed to diffuse freely throughout the tissue, and chemical reactions are suppressed; and SWITCH-ON, when the buffer environment is rapidly changed to a condition allowing chemical reactivity. Tissue architecture, native biomolecules, and antigenicity are highly preserved in this method, allowing for multiple rounds (>20) of labeling.
C. DiD staining of a 1-mm-thick mouse coronal block using PBST for 1.5 days at 37°C. Only tissue surface is labeled. Scale bar, 200 μm.
D. DiD staining of a 1-mm-thick mouse coronal block using SWITCH. The sample was first incubated in DiD, 10 mM SDS containing PBS buffer for 24 hr, then moved to PBST and incubated for 0.5 day at 37°C. The whole sample is uniformly labeled. Scale bar, 200 μm.
Stochastic Electrotransport (SE), published in PNAS in 2015, is a novel method for rapid, nondestructive processing of porous samples. Processes like delipidation and immunolabeling have historically been limited by diffusion time, particularly for large samples such as an intact mouse brain. It may require weeks for antibodies to diffuse through only a few millimeters of tissue, and the labeling gradient from surface to core can be steep. SE uses a rotational electric field to disperse highly electromobile molecules (such as antibodies or surfactant micelles) throughout a porous sample without damaging electrically charged structures within the tissue. This method enables rapid clearing (2-4 days) of intact organs and rapid staining (<24 hours) with nuclear dyes, proteins, and antibodies with incredible uniformity.
