Azide (Az)/Difluorocyclooctene (DIFO)-based Click Imaging Service
Your Trusted Azide (Az)/Difluorocyclooctene (DIFO)-based Click Imaging Partner
Az/DIFO-based click imaging is a bioimaging technique used to detect interactions between biomolecules. It uses the chemical reaction between two small molecules, Az and DIFO, to label them on the biomolecule, and then observe their location and quantity in cells or tissues. CD BioGlyco has extensive experience with GlycoCLICK™-based Imaging Services to ensure results are delivered on time. Our scientific project managers provide individual oversight of each project to support your Az/DIFO-based click imaging project. Here are the details of our service:
- Labeling strategies
We achieve Az/DIFO imaging by using different labeling strategies. Our labeling strategies are divided into direct and indirect labeling methods. The direct labeling is to bind the Az/DIFO directly to the target molecule to form a fluorescent probe for imaging. Indirect click-labeling utilizes Az and DIFO that bind to a specific linker (e.g., click-responsive complement), which then reacts with the target molecule. This method can be extended to apply to more types of target molecules because the linker can be selected to fit the pairing reaction of a specific target molecule. With the two-step reaction, Az and DIFO can be introduced to the target molecule, enabling imaging of the target molecule.
- Imaging methods
We not only provide Az/DIFO imaging technology but also offer to combine it with different imaging methods to observe and document the localization of molecules within cells. Our commonly used imaging methods include fluorescence microscopy, confocal laser scanning microscopy (CLSM), structured light microscopy, and multiphoton microscopy. Since these methods exhibit different resolutions and imaging depths, we select the appropriate method based on your specific research needs.
- Image processing and analysis
We process the acquired fluorescence microscope images by applying image processing software and algorithms for denoising, enhancing, and segmenting to obtain clear images. We then use relevant analytical methods to quantitatively measure the intensity, distribution pattern, and co-localization relationship of the fluorescence signals to obtain quantitative data about molecular localization.
Fig.1 Process of Az/DIFO-based click imaging. (CD BioGlyco)
Publication Data
Technology: Metabolic labeling and western blot
Journal: ACS Chemical Biology
IF: 4.0
Published: 2009
Results: This article focuses on the in vivo imaging of glycans in Caenorhabditis elegans (C. elegans). Specifically, the researchers used a method called "glycan metabolic labeling" to image glycans in vivo by adding specific glycan labels to C. elegans to make them glow. The authors introduced azidosugar-containing glycan molecules into C. elegans by metabolic labeling. These azidosugars were processed and incorporated into newly synthesized glycan molecules through the action of intracellular esterases, a portion of which is displayed on cell surface glycoproteins. The authors used a fluorescent probe containing DIFO to react with metabolically labeled azidosugar in C. elegans. This reaction does not require a copper catalyst and can be performed in vivo. Finally, protein blot analysis was performed to detect glycoproteins containing azide labels. This method could help researchers understand the distribution and function of glycans in C. elegans.
Fig.2 Strategy for in vivo imaging of glycans in C. elegans. (Laughlin & Bertozzi, 2009)
Applications
- This technology is applied to study the detection and localization of tumor markers by attaching Az/DIFO labeling to tumor markers, allowing for their efficient visualization and localization in living cells and tissues.
- This imaging technique visualizes and localizes proteins by attaching Az/DIFO small molecule labeling to target proteins through specific chemical reactions, and allows for the study of protein interactions.
- Click imaging technology allows visualization and localization of drugs and targets by introducing Az/DIFO labeling into a drug or drug candidate compound.
Advantages
- Our Az/DIFO-based imaging technology can be combined with other imaging methods and techniques to provide more comprehensive and multidimensional information.
- This technique is highly selective, allowing researchers to accurately visualize and localize biomolecules of interest, such as proteins and nucleic acids, without interfering with the observation of other molecules.
- The click reaction between Az and DIFO is highly efficient and the reaction product has a strong fluorescence or emission signal so that the target molecule can be detected at low concentrations.
At CD BioGlyco, we provide internationally acclaimed scientific project management that assists you throughout the entire process of your customized Az/DIFO-based click imaging project, starting from design and concluding with delivery. If you are interested in our services, please feel free to contact us.
Reference
- Laughlin, S.T.; Bertozzi, C.R. In vivo imaging of Caenorhabditis elegans glycans. ACS chemical biology. 2009, 4(12): 1068-1072.
For research use only. Not intended for any clinical use.
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