Fluorescence Labeling

Fluorescence Labeling of Peptides

Fluorescence Labeling of Oligonucleotides

Fluorescence Labeling of Antibody

Fluorescence Labeling of Proteins

Fluorescence Labeling of Nucleic Acids

Fluorescence Labeling Polysaccharide

Fluorescence Labeling of Enzyme

Fluorescence Labeling Carbohydrate

Fluorescence Labeling Fatty Acid

Fluorescence Labeling of Magnetic Nanoparticles

Fluorescence labeling is a technique used in biological and medical research to mark or tag specific proteins, cells, or other molecules with a fluorescent substance (known as a fluorophore). As a professional biotech company, we provide top-notch services related to the technology of fluorescence labeling. This is aimed at facilitating the progress of scientific research and supporting the collective mission of the worldwide scientific fraternity towards enhancing human health.

What are fluorescent substances?

Fluorescent substances are compounds on which fluorescent labeling depends. Fluorescent substances are compounds with the chemical structure of conjugated double bond systems, which can be transformed to the excited state when irradiated by UV light or blue-violet light and emit fluorescence when restoring to the ground state.

Fluorescent labeling technique originated in the 1940s and was primarily used for antibody labeling to detect the corresponding antigens. It involves binding a fluorescent substance, either covalently or physically, to molecule groups. This method is employed in various research studies such as nucleic acid, protein, and peptide, to gather information about the subject. Under this technique, a fluorescent dye or tag is affixed to the interest target, and with the help of special devices that emit and detect light, it can be tracked and visually represented. The tagged target emits light when subjected to a specific light wavelength, enabling scientists to monitor its location, movement, or interaction within a system. This methodology is extensively utilized in numerous fields such as molecular biology, microbiology, medical diagnostics, environmental science, and drug discovery. It proves to be especially beneficial in researching biological activities at the molecular and cellular level.

Fig.1 PEG Fluorescent Labeling

Common Reagents for Fluorescence Labeling

Advantages of Fluorescence Labeling

Sensitivity: Fluorescence labeling is incredibly sensitive, with the ability to detect and quantify minute amounts of a sample. The sensitivity allows for less sample material and reagents to be used, enhancing cost-effectiveness.

Specificity: Fluorescent tags can be designed to bind to specific molecular targets within the cell, providing highly specific labeling. This results in a high signal-to-noise ratio, making it much easier to detect specific molecules.

Multiplexing: Using different colored fluorescent labels allows researchers to detect multiple targets within the same sample simultaneously. This is known as multiplexing, and it allows complex biological systems to be studied more comprehensively.

Non-destructive: Fluorescence labeling is non-destructive, which means that it does not damage or alter the samples. The sample can be analyzed multiple times, and further used for other experiments.

Real-time Monitoring: It allows real-time observation of biological processes within live cells. Scientists can watch how molecules interact and change over time adding to the understanding of molecular and cellular dynamics.

Quantification: Fluorescence labeling allows quantification of cells or molecules, which can be valuable for measuring gene expression, protein-protein interaction, etc.

Versatility: Fluorescent labels can be used in diverse applications, from in vitro experiments like flow cytometry and immunofluorescence to in vivo imaging of experimental animals.

Improved Contrast: Fluorescent tags enhance contrast in microscopy, making it easier to visualize samples compared with conventional light microscopy.

No Radioactivity: Unlike some other methods of labeling, fluorescence does not involve hazardous radioactive substances, making it safer for both the experimenter and the environment.

Applications of Fluorescence Labeling

Medical Diagnostics: Fluorescence labeling is commonly used in medical diagnostics such as immunoassays to highlight the presence of specific proteins or other substances in body fluids.

Drug Discovery: In pharmaceutical research, fluorescence labeling is often used to monitor drug distribution within a biological system. It helps in understanding how a drug interacts within the body and how it is distributed across various cells.

Cell Biology: This technique is widely used in cell biology to track the movement and interaction of cells and cell components. It helps in understanding the mechanisms of cell division, protein synthesis, and more.

Genetic Analysis: Fluorescence labeling can be used to mark specific DNA or RNA sequences, allowing for the identification of genetic markers linked with particular diseases.

Microscopy: Fluorescence labeling is a crucial tool in fluorescence microscopy, where it allows for the visualization of specific structures within cells or tissues.

Biochemical Assays: Fluorescence labeling is commonly used in biochemistry for binding studies and enzymatic assays. It can help detect molecular interactions, enzyme activity, and concentration of various biomolecules.

Environmental Monitoring: Fluorescence labeling can be used to track pollutants or contaminants in the environment, helping with the monitoring and management of environmental health.

Food Industry: This technique can be used for the detection of microorganisms or pathogens in food and beverage samples, ensuring food safety.

Nano Technology: Fluorescence labeling can be used in the field of nanotechnology to track nanoparticles in various environments.

Cancer Research: Fluorescence labels can make tumor cells visible, enabling better diagnosis and treatment of cancer. Fluorescent dyes can help identify cancerous cells or can be used to deliver drugs precisely to cancer cells, reducing side effects.

Our Fluorescence Labeling Services

Fluorescent Dye Labeling: We can conjugate fluorescent dyes to various biomolecules such as proteins, antibodies, peptides and nucleic acids. These can then be used for research or diagnostic applications.

Custom Labeling: Based on the specific requirements of our clients, we can customize the fluorescence labeling process. This encompasses the selection of the appropriate dye, optimization of labeling conditions to maintain the biological function of the molecule and increase its stability.

Fluorescence Immunoassay Development: Our services also include developing and optimizing fluorescence immunoassays for the detection and quantification of a wide range of targets.

Bioconjugation Services: We have expertise in bioconjugation techniques and can conjugate fluorescent tags to a variety of biological molecules.

FRET (Fluorescence Resonance Energy Transfer) Studies: We perform FRET studies to probe protein interactions, conformational changes and other biochemical phenomena.

Fluorescence Microscopy Imaging: We utilize advanced microscopy techniques for the characterization and visualization of labeled biomolecules.

Consulting & Training: Our team of experts can provide consultation and training on fluorescence labeling techniques, their applications, and interpretation of results.

Comprehensive Analytical Support: Post-labeling, we provide comprehensive analysis, including the identification of labeling sites, determination of the degree of labeling, and validation of the physicochemical and biological properties of the labeled molecule.

To summarize, we are dedicated to delivering state-of-the-art fluorescence labeling services and solutions tailored to the unique needs of your research studies, diagnostic assay development or therapeutic applications.

General Workflow for Fluorescence Labeling

Sample Preparation: This includes selection and gathering of the right sample for the experiment. The samples can be biological cells, tissues, proteins, etc. The samples are usually kept in conditions that maintain their integrity and biological function.

Initial Cleaning of the Sample: It's important to pre-clean the sample to remove any possible contaminants that could interfere with the labeling process.

Selection of Fluorescent Dyes: Choose the suitable fluorescent dye depending on the experiment's goal and the nature of the sample. Each fluorescent dye has its own excitation and emission properties, cellular retention capacity, and biological compatibility.

Fluorescent Labeling: In this step, the dye is attached to (or incorporated into) the molecules of interest (targets). The attachment can be through chemical reactions or merely just mixing the target substances and fluorescent dyes under an appropriate environment to allow the binding to happen.

Washing and Purification: After the labeling process, the sample could be washed to remove any unbound fluorescent dye. This might sometimes include a purification process to further remove the unbound and loosely bound dye molecules.

Validation: Once the fluorescent labeling has been done, the samples might be subject to validation processes like checking under a microscope to observe if the labeling has been successfully done.

Observation and Data Gathering: Fluorescence Microscope, Flow cytometer, or another suitable equipment is used to observe and detect the fluorescence. The fluorescence intensity reflects the concentration or localization of molecules of interest.

Data Analysis and Interpretation: After gathering the data, analysis will be performed to make sense of the observations. The interpretations will be made based on the experiment objectives.

Reporting and Publication: Finally, the results of the experiment are reported for interpretation, reviewed, and possibly published in academic journals. The data should be reproducible and interpretable by others in the field.

Re-Iteration: Based on the results or feedback, it's common to then refine the experiment or move onto additional experiments to answer more questions.

Remember to always maintain safety measures while performing the experiment as some fluorescent dyes can be hazardous.

Existing Fluorescence Labeling Techniques

Competitive Advantages

High Expertise: Our team is composed of experienced professionals and researchers who are masters in the field of fluorescence labeling technology. Their outstanding skills and deep knowledge base guarantee exceptional service delivery.

Advanced Technologies: We use state-of-the-art technologies in fluorescence labeling that support the highest level of precision and accuracy. Our techniques facilitate efficient bioconjugation of fluorescent dyes to various biomolecules.

Cost-Effective Solutions: Despite offering high-end services, our pricing structure is designed to be affordable. We aim to provide more individuals and institutions access to quality fluorescence labeling technology.

Quick Turnaround: Our efficient procedures and diligent team members ensure that we meet all timeliness requirements set by our clients.

Customizable Services: Understanding that different clients have unique needs, we offer personalized fluorescence labeling services to meet those specific needs.

Excellent Customer Support: We have a prompt and friendly customer support team ready to answer any queries and resolve any issues that may arise in your interaction with our service.

In conclusion, our company provides a quality, cost-effective and efficient solution for Fluorescence Labeling technology services, backed by a skilled team providing excellent customer service.

Fluorescence Labeling Protocol

Materials:

Sample

Fluorescent dye or label

Suitable solvent

Specific tools or equipment such as a fluorescence microscope or flow cytometer

Procedure:

• Choose the appropriate fluorescent dye or labels: These labels must be appropriate for your type of sample and should specifically bind or attach to the molecule or structure you are interested in. Some may require a specific binding partner to be present in the sample.
• Adding the fluorescent label to your sample: Dissolve your label in a suitable solvent, and then add it to your sample. Depending on the type of label and sample, you might need to incubate your sample for some time at a certain temperature or in specific conditions.
• Washing and removing excess labels: This step is very crucial, as you need to remove any unattached or excess fluorescent labels that can give false-positive results. This is usually done by washing your sample multiple times with a suitable buffer.
• Observation and data collection: Your labeled sample is now ready to be observed under a fluorescence microscope or analyzed using a flow cytometer, where the light emitted from the excited fluorescent labels will be captured and analyzed.
• Data analysis: The data captured will be further analyzed using relevant software to draw conclusions.

Remember to always run a negative control (a sample with no fluorescent label) along with your test samples.

Notes: This is a general protocol and depending on the type of sample (cells, tissues, biomolecules, etc.) and the type of fluorescent labels used, specific methods may vary. Always refer to the label's or kit's user manual and best practices in your field.

Case Study

• Case Study 1

The process of fluorescence labeling DNAs is critical yet the existing techniques tend to be costly and labor-intensive. We introduce a novel, efficient, and affordable method for fluorescence labeling of oligonucleotides using base excision trapping (BETr). This method utilizes deaminated DNA bases to identify label positions and aminooxy-substituted rotor dyes to highlight AP sites, leading to high emission intensities. Importantly, BETr can work in conjunction with DNA synthesis by polymerases, allowing for the integration of multiple uracil into an amplicon and in situ BETr labeling without any need for washing. Furthermore, BETr permits high labeling density of dsDNA such as genomic DNA in a single tube through nick translation. By using two different deaminated bases, two-color site-specific labeling is achieved. This methodology also utilizes a multi-labeled DNA construct as a luminescent tag, an added advantage demonstrated by linking to an antibody for protein imaging. The technique also leverages the double-strand selectivity of a repair enzyme in accurately detecting the presence of a target DNA or RNA in a mixed sample with isothermal turnover and single nucleotide specificity. Hence, BETr offers a highly adaptable and easy way for general fluorescence labeling of DNAs.

Fig.2 In situ DNA synthesis and BETr labeling. (Jun, Y.W., 2022)

• Case Study 2

The recent notable rise in the usage of deep eutectic solvents (DESs) offers novel methodologies for sample collection and preparation. However, utilizing these new solvents poses difficulties for subsequent analyses since standard analytical techniques, like fluorescence labeling, may be incompatible. This study investigates the efficacy of three conventional fluorescent labels in derivatizing amino acids in prevalent DESs, comprised of choline chloride and ethylene glycol. We found that the distinctive solvent attributes of the DESs permit two fluorophores, fluorescein isothiocyanate and 5-carboxyfluorescein succinimidyl ester, to successfully label amino acids. Conclusions indicate the feasibility of effectively labeling both D- and L-amino acids at concentrations as low as 4 μM in solution. Following this process, capillary electrophoretic separation is able to detect as low as 50 nM, without the need to eliminate any DES. This is the first time a total fluorescent labeling reaction has been carried out in a DES, followed by capillary electrophoretic separation of the analytes.

Fig.3 Representative electropherograms of various amino acids simultaneously reacted in 5 mM KOH ethaline at (1) Leu, (2) L-His, (3) L-Ser, (4) L-Ala, (5) Gly, and (6) L-Glu, all at 25 μM amino acid with FITC. (2. Torres, J., 2022)

FAQ

1. What is fluorescence labeling?

Fluorescence labeling is a technique used in biological and medical research to tag or highlight specific proteins, cells, or other molecules of interest. This is achieved by attaching fluorescent dyes or proteins to the target molecules, which emits light of a longer wavelength when excited by light of a shorter.

1. How does fluorescence labeling work?

The process of fluorescence labeling involves attaching fluorescent dyes or proteins to the target molecules. Once these molecules are subjected to light of a certain wavelength, they absorb the light energy and re-emit it as light of a longer wavelength, producing a glow that can be detected and analyzed.

2. What is fluorescence labeling of DNA?

Fluorescence labeling of DNA is a technique used in molecular biology where fluorescent dyes are attached to DNA molecules. These labeled DNA fragments can then be visualized under a microscope or in an analytical instrument through the generation of a fluorescent signal. This method is commonly used in various DNA tests and experiments including gene mapping, DNA sequencing, DNA microarrays, and studies involving gene expression and regulation.

3. How does DNA fluorescence work?

It works on the principle that certain chemicals, known as fluorescent dyes, can intercalate (slot between) the bases (A, T, C, G) of DNA. When these dyes are exposed to a specific wavelength of light, they absorb the energy and enter into an excited state. After a short period, they drop back down to their normal energy state and give off a photon - this is the fluorescence that can be detected.

4. What are the advantages of using fluorescence labeling?

This technique allows for the visualization and analysis of specific biological substances in a much clearer and more detailed manner than traditional methods. It aids in studying cellular processes, protein-protein interactions and the distribution of molecules within cells. It is highly sensitive, precise, and can be used in live cells and organisms.

5. In what fields can fluorescence labeling be used?

Fluorescence labeling can be utilized in many biological and medical research fields. This encompasses cell biology, microbiology, immunology, genetics, biochemistry, and more. They are frequently used in diagnostic assays, drug discovery and development, as well as in medical imaging procedures.

6. Are there any risks or side effects of fluorescence labeling?

The main challenge of fluorescence labeling is to introduce the fluorescent label without interfering with the function of the molecule or system of interest. Some fluorescent tags can induce harmful effects when exposed to light, potentially causing cellular damage. Therefore, careful calibration and handling procedures are crucial in the use of this technique.

7. How much does fluorescence labeling service cost?

The cost of fluorescence labeling service can vary greatly depending on the type and amount of material to be labeled, the complexity of the procedure, and the specific fluorescent label used. Please contact us directly for a more specific and accurate quotation.