Fluorophore-based Liposome Fusion Cell Surface Glycoengineering Service

Fluorophore-based Liposome Fusion Technology

Fluorophore-based liposome fusion technology represents a potent approach to investigating membrane fusion processes. This technique relies on fluorophores for the labeling of liposomes and target membranes, enabling real-time monitoring of fusion events via changes in fluorescence. It finds valuable applications in fields such as drug delivery, cellular research, and the exploration of membrane interactions.

This technology is particularly useful for a protocol designed to assess the kinetics of fusion stages and visualize fusion complexes involving influenza viruses and liposomes. This is achieved through fluorescence dequenching assays and cryo-electron tomography (cryo-ET). Fluorescence dequenching assays allow the tracking of membrane fusion progress by monitoring factors like liposomal content leakage and lipid mixing. Two commonly employed dyes in this protocol are sulforhodamine-B (SRB) and the lipophilic fluorophore DiD.

Fig.1 Flowchart of the major steps in the preparation of (A) SRB-labeled liposomes and (B) DiD-labeled influenza viruses. (Gui & Lee, 2018)

Fluorophore-based Liposome Fusion Cell Surface Glycoengineering Service at CD BioGlyco

CD BioGlyco provides custom fluorophore-based liposome fusion cell surface glycoengineering services for clients, fluorophores used for liposome fusion cell surface glycoengineering should be selected based on compatibility with the experiment's goals and the specific glycan or cell surface modification. According to the client's needs, we offer a suitable solution for clients to bring the professional protocol and help our clients study the field of glycobiology. Moreover, CD BioGlyco also provides other Liposome Fusion-based Cell Surface Glycoengineering Services such as the GAGs-based Liposome Fusion Cell Surface Glycoengineering Service and the Hydrophobic Membrane Insertion-based Cell Surface Glycoengineering Service. These services can be comminated with other Technologies for Cell Surface Glycoengineering to help researchers study the potential applications of Cell Surface Glycoengineering.

Fig.2 The typical steps in a fluorophore-based liposome fusion process. (CD BioGlyco)

Several dyes are commonly used for lipid labeling in various biological and biochemical applications.

DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate): DiI is a lipophilic carbocyanine dye used for long-term cell tracing and lipid membrane labeling.
DiO (3,3'-dioctadecyloxacarbocyanine perchlorate): DiO is another carbocyanine dye ideal for lipid membrane labeling and cell tracing.
Nile Red: Nile Red is a hydrophobic, lipophilic dye used for lipid staining and labeling. It exhibits strong fluorescence in nonpolar environments.
FM Dyes (styryl pyridinium dyes): FM dyes like FM1-43 are used for labeling synaptic vesicles and endocytic pathways. They are particularly useful for live-cell imaging.
BODIPY Dyes: Various BODIPY dyes are suitable for lipid labeling. They come in different emission wavelengths and have diverse applications.
Laurdan: Laurdan is used as a probe for the polarity of lipid bilayers. It undergoes a shift in the emission spectrum depending on the polarity of the environment.
Fluorescein-Phosphatidylethanolamine (fluorescein-PE): This compound is a green fluorescent phospholipid used for membrane labeling.
Fluorescent Cholesterol Derivatives: Cholesterol derivatives, such as NBD-Cholesterol, are used to label membrane cholesterol, an important lipid component.
Dansyl-DHPE: Dansyl-DHPE is used for labeling and studying membrane properties.

Publication

Publication Data

Paper Title: Directing neuronal signaling through cell-surface glycan engineering.

Technology: Membrane fusion, liposomes, fluorescence spectrometry, and cryo-electron tomography.

Book Series: Methods in Molecular Biology

IF: NA

Published: 2018

Results: In this research, scientists outline a methodology for investigating the fusion of viruses and liposomes. They employ a combination of fluorescence dequenching assays and cryo-electron tomography (cryo-ET). Fluorescence dequenching, in particular, evaluates membrane fusion effectiveness by monitoring entire influenza viruses tagged with a lipophilic dye (DiD) and liposomes labeled with a water-soluble dye (sulforhodamine B). Simultaneously tracking these fluorescent signals allows for determining the relative timeframes of liposomal content leakage or transfer compared to lipid merging. Additionally, cryo-ET provides the capability to capture three-dimensional images of various fusion stages, including pre-fusion, fusion intermediates, and post-fusion events.

Here are some of the results shown in this research.

Fig.3 Schematic diagram shows fluorescence dequenching assays. (Gui & Lee, 2018)

Applications

Drug Delivery: This technology can be employed to design liposomes that carry therapeutic agents, allowing for targeted drug delivery to specific cells or tissues.
Cellular Studies: Researchers can use this approach to investigate cell membrane dynamics, cellular interactions, and fusion events, providing insights into fundamental cellular processes.
Virus-Host Interactions: It enables the study of virus-cell membrane fusion events, shedding light on viral entry mechanisms and facilitating the development of antiviral strategies.

Highlights

We have a well-equipment Glycoengineering Platform and provide high-quality fluorophore-based liposome fusion glycoengineering service for our clients.
We offer custom solutions for our clients and aim to be an outstanding company in the field of glycobiology.
Our talented scientists and well-experienced laboratory team can perform our experiments and subsequent data analysis in good quality, providing our clients with reliable laboratory reports.

CD BioGlyco has adventure experience in the field of glycobiology and has established an excellent glycoengineering platform to offer several custom cell surface engineering services. If you are interested in our services and would like to get in touch with us, please feel free to contact us for more information.

Reference

Gui, L.; Lee, K.K. Influenza virus-liposome fusion studies using fluorescence Dequenching and cryo-electron tomography. Methods in Molecular Biology. 2018, 261–279.

This service is for Research Use Only, not intended for any clinical use.