Flow Cytometry
BOC Sciences remains committed to delivering top-notch biochemical products and services, which include high-quality fluorescent dyes alongside essential support services. Our fluorescent dye selection covers traditional organic dyes as well as fluorescent proteins and innovative fluorescent nanomaterials to serve various flow cytometry application demands. Our products provide superior spectral properties along with high fluorescence intensity and stability while maintaining biocompatibility, which makes them dependable for various research applications such as cell surface labeling, intracellular staining, and multicolor analysis. In addition, BOC Sciences provides tailor-made fluorescent labeling support, enabling clients to speed up their research while achieving better efficiency in cell analysis.
What is Flow Cytometry?
Flow cytometry serves as a high-throughput method for analyzing and sorting cells. The system operates by directing a cell suspension through a restricted flow chamber, which aligns cells individually before they traverse a laser beam. Cells emit specific wavelength fluorescence signals when they pass through a laser because the fluorescent dyes inside them become excited. Detectors capture these signals, which they convert into electrical signals for computer analysis to determine cell parameters like size, shape, internal structure, and surface markers. The technique of flow cytometry enables scientists to count cells and analyze phenotypes while simultaneously studying cell functions and sorting cells based on cycle analysis. Flow cytometry serves multiple applications in basic research, clinical diagnostics, and drug development because of its high throughput and sensitive multi-parameter analysis capabilities.
Fig. 1. Schematic diagram of a flow cytometer 1,2 .Comprehensive Fluorescent Dye Technical Support: Core Advantages of BOC Sciences
Strong R&D Team
We have a cross-disciplinary team of chemists, biologists, and engineers who can quickly respond to customer needs and develop high-performance fluorescent dyes.Advanced Synthesis Technology
We utilize advanced organic synthesis and bio-conjugation techniques to ensure the high purity and high activity of fluorescent dyes.Strict Quality Control
Every batch of fluorescent dyes undergoes rigorous quality testing to ensure their performance meets customer requirements.Rich Product Line
We offer a variety of fluorescent dyes to meet a wide range of application needs and can customize synthesis according to specific customer requirements.
Boost Your Experiments with Our Top-Tier Fluorescent Dyes
BOC Sciences provides various types of fluorescent dyes for flow cytometry to meet different research and application needs. These dyes offer high sensitivity, low background noise, and excellent photostability, making them suitable for multiplex labeling and complex sample analysis.
Organic Fluorescent Dyes
- FITC
- Phycoerythrin (PE)
- PerCP
- Allophycocyanin (APC)
- Alexa Fluor Series
- Cy5, Cy7
- BODIPY
- Coumarin
Fluorescent Proteins
- Green Fluorescent Protein
- Red Fluorescent Protein
- Yellow Fluorescent Protein
- Cyan Fluorescent Protein
Tandem Dyes
- PE-Cy5
- PE-Cy7
- APC-Cy7
- PerCP- Cy5.5
Nucleic Acid Dyes
- Hoechst 33342
- 7-AAD
- DAPI
- PI
One-Stop Custom Synthesis and Modification Service of Fluorescent Dyes
BOC Sciences fully understands the diverse needs for fluorescent dyes in scientific research and industrial applications. Different experimental systems and detection platforms often require fluorescent labels with specific wavelengths, specificity, and sensitivity. To meet these needs, BOC Sciences offers highly flexible customized fluorescent dye services, covering the entire process from molecular structure design, chemical synthesis, conjugation labeling to application optimization, to fulfill various experimental requirements in flow cytometry and related fields. Through in-depth communication with clients and personalized design, BOC Sciences ensures that each customized fluorescent dye exhibits excellent performance and reliability.
Personalized Structure Design
The chemical structure of fluorescent dyes directly impacts their optical performance and biocompatibility. BOC Sciences' scientists design personalized fluorophore structures for clients based on the chemical properties of target molecules and excitation/emission wavelength requirements. For example, by modifying electron-donating or accepting groups, the emission wavelength of fluorescent dyes can be adjusted to meet the needs of multicolor fluorescence detection.
Spectral Property Optimization
For multiplex fluorescence labeling experiments, BOC Sciences can optimize the excitation and emission wavelengths of dyes based on the spectral characteristics of different fluorescence channels to avoid fluorescence overlap and improve signal resolution. This service is especially useful for high-throughput analysis of complex cell populations or subpopulations.
Multifunctional Modification
BOC Sciences can introduce various functional groups onto fluorescent dyes, such as carboxyl, amino, maleimide, and azide groups, enabling efficient covalent coupling with proteins, nucleic acids, sugars, and other biomolecules. This multifunctional modification not only enhances the stability and specificity of the labeling but also expands the application range of fluorescent dyes, such as for bioorthogonal labeling in Click chemistry reactions.
Solubility and Stability Regulation
For different experimental environments (such as aqueous solutions, organic solvents, or biological buffers), BOC Sciences can regulate the solubility and photostability of fluorescent dyes through molecular design and chemical modification, thereby improving the reproducibility and data quality of experiments.
Precise and Efficient Fluorescent Labeling Services from Antibodies to Nucleic Acids
BOC Sciences provides efficient and precise fluorescent labeling services, covering antibodies, proteins, peptides, and nucleic acids. These services help clients save time and effort while ensuring the high quality and consistency of the labeled products.
Antibody Labeling
Fluorescently labeled antibodies are among the most commonly used experimental tools in flow cytometry. BOC Sciences employs advanced labeling techniques, such as N-hydroxysuccinimide (NHS) ester and maleimide chemical conjugation, to efficiently covalently bind fluorescent dyes to lysine or cysteine residues on antibodies. The labeled antibodies retain their efficient antigen-binding ability while possessing high fluorescence intensity and signal-to-noise ratio, ensuring sensitivity and accuracy in detection. For multicolor flow cytometry, BOC Sciences offers fluorescence spectral analysis and pairing optimization services to help clients select the most compatible fluorescent dye combinations, minimizing channel overlap and crosstalk.
Protein Labeling and Peptide Labeling
BOC Sciences selects appropriate labeling sites and fluorescent dyes based on the structural features of proteins or peptides, ensuring that the conjugation of the fluorophore does not affect the biological activity of the target molecule. Efficient conjugation chemistry and professional purification techniques (such as gel filtration, ion exchange, and affinity purification) are employed to ensure labeling efficiency and purity. Labeled proteins and peptides can be used for intracellular dynamics research, protein-protein interaction analysis, and enzyme activity detection.
Nucleic Acid Labeling
For DNA and RNA fluorescent labeling, BOC Sciences offers a variety of compatible fluorescent dyes and labeling strategies, such as coupling amino-modified oligonucleotides with NHS fluorescent dyes and Click chemistry labeling methods. Depending on nucleic acid sequences and experimental needs, BOC Sciences provides several labeling strategies, including end-labeling, internal site labeling, and multiplex fluorescence labeling. The labeled probes exhibit high specificity and sensitivity, making them suitable for a variety of molecular biology experiments, such as FISH, qPCR, and RNA detection.
Cell Labeling
BOC Sciences provides comprehensive fluorescent cell labeling services, covering cell membranes, organelles, cell cycles, and cell viability and apoptosis markers. BOC Sciences also offers fluorescent labeling for cell activity, proliferation, and functional status, such as: labeling cells with CFSE or CellTrace to track cell division and proliferation dynamics; using fluorescent probes like Fura-2, Fluo-4, and DCFDA to monitor intracellular calcium signals, pH changes, and oxidative stress states, revealing mechanisms of cell signaling and metabolic regulation.
Stringent Quality Testing Ensures the Reliability of Fluorescent Dyes
Fig. 2. Analytical instruments and equipment (BOC Sciences Authorized).
- Spectral Analysis: Using advanced spectrophotometers, the absorption and emission spectra of dyes are tested to ensure that they have optimal fluorescent properties at specific wavelengths. These spectral data help verify the purity and fluorescence efficiency of the dyes.
- Purity Testing: The purity of the dyes is tested using technologies such as high-performance liquid chromatography (HPLC), ensuring that no impurities are present, thereby guaranteeing the accuracy of experimental results.
- Stability Testing: Fluorescent dyes undergo thermal, photostability, and chemical stability tests to ensure that their performance does not degrade during long-term storage and use.
- Particle Size Testing: For nanoparticle-based fluorescent dyes, dynamic light scattering (DLS) technology is used to analyze particle size distribution, ensuring uniformity and the required size range.
- Biocompatibility and Cytotoxicity Testing: To ensure the safety of fluorescent dyes in cell experiments and in vivo applications, we employ rigorous biocompatibility and cytotoxicity testing, including MTT assays, apoptosis detection, and activity evaluation.
- Batch-to-Batch Consistency Testing: By conducting comparative tests on different batches of fluorescent dyes, we ensure product performance consistency, preventing batch-to-batch variations from affecting experimental results.
Advanced Analytical Platform
- UV-Vis Spectrophotometer
- FTIR
- NMR
- Fluorescence Spectroscopy
- HPLC
- GC
- GPC
- TLC
- LC-MS
- GC-MS
- AAS
- ICP-MS
- XRF
- DSC
- TGA
- Melting Point Apparatus
- Polarimeter
- Viscometer
- XRD
- Karl Fischer Titration
Diverse Applications of Fluorescent Dyes in Flow Cytometry
Immunophenotyping
By conjugating fluorescent dyes with specific antibodies, researchers can simultaneously detect the expression of multiple antigens on the surface or inside cells. This multiplexing technique allows for fine differentiation of immune cell subsets, such as T cells, B cells, NK cells, and their functional status.
Cell Cycle and Apoptosis Analysis
Fluorescent dyes are indispensable in cell cycle and apoptosis analysis. Nucleic acid dyes like PI and DAPI bind to DNA, enabling quantitative measurement of DNA content and cell cycle distribution (G0/G1, S, G2/M phases). Additionally, fluorescent dyes such as Annexin V-FITC detect the externalization of phosphatidylserine during early apoptosis.
Intracellular Calcium Detection
Fluorescent dyes offer unique advantages for intracellular calcium detection. Calcium indicators like Fluo-3 and Fura-2 bind specifically to calcium ions and undergo fluorescence intensity changes upon binding. Using flow cytometry, researchers can monitor real-time dynamic changes in intracellular calcium concentrations and study cellular signaling mechanisms.
Reactive Oxygen Species (ROS) Detection
Reactive oxygen species (ROS) play an essential role in cellular physiology and pathology. Fluorescent dyes like DCFH-DA and HE react with ROS to produce fluorescence signals, enabling the detection of oxidative stress levels in cells. Using flow cytometry, researchers can quantify ROS production and clearance, studying their role in disease progression.
Cell Sorting
Fluorescent dyes play a key role in cell sorting. By binding fluorescent dyes to specific surface or intracellular markers, researchers can use flow cytometry to isolate target cells. For example, stem cells, immune cells, or cancer cells can be isolated and enriched through specific labeling.
Drug Screening
Fluorescent dyes are crucial in high-throughput drug screening. By conjugating fluorescent dyes with cells or molecules, researchers can quickly assess drug effects, such as cytotoxicity, apoptosis induction, or signal pathway activation, using flow cytometry.
FAQs About Flow Cytometry
What is flow cytometry test?
Flow cytometry is a technique used to analyze and sort cells. It involves injecting a cell suspension into a flow chamber, where cells pass through a laser beam and scatter light and emit fluorescence signals. These signals reflect the cell’s size, shape, and characteristics of surface and intracellular molecules. Flow cytometry is widely used in immunology, cancer research, and hematology to analyze large cell populations and obtain multiparameter data.
What is flow cytometry used for?
Flow cytometry is primarily used for cell analysis and sorting. It can detect proteins on the cell surface, DNA content, cell cycle state, and more, with applications in immunophenotyping, cancer diagnostics, stem cell research, and microbiological detection. Additionally, flow cytometry is used for functional studies such as apoptosis, proliferation, and calcium concentration measurement, providing critical tools for biomedical research.
What dyes are used in flow cytometry?
Common dyes used in flow cytometry include fluorescent dyes and nucleic acid dyes. Fluorescent dyes like FITC, PE, and APC are used to label antibodies for detecting proteins on the cell surface or inside cells. Nucleic acid dyes such as PI (propidium iodide), DAPI, and Hoechst 33342 are used to measure DNA content and analyze the cell cycle. There are also functional dyes, like calcium ion indicators and ROS detection dyes, used to study cell functions.
What are fluorophores in flow cytometry?
Fluorophores are fluorescent substances used in flow cytometry to label cells or molecules. They absorb light at a specific wavelength and emit fluorescence at a different wavelength, allowing detection by sensors. Common fluorophores include organic dyes (e.g., FITC, PE), protein fluorophores (e.g., phycoerythrin), and small molecule fluorophores (e.g., Alexa Fluor series). The choice of fluorophore depends on experimental needs such as excitation wavelength, emission wavelength, and brightness.
References
- Image retrieved from Figure 1 " Schematic diagram of a flow cytometer, showing focusing of the fluid sheath, laser, optics (in simplified form, omitting focusing), photomultiplier tubes (PMTs), analogue-to-digital converter, and analysis workstation. https://doi.org/10.1371/journal.pcbi.1003365.g001," O'Neill et al. , 2013, used under [CC BY 4.0](https://creativecommons.org/licenses/by/4.0/). The title was changed to " Schematic diagram of a flow cytometer".
- O'Neill, K. et al. Flow cytometry bioinformatics. PLoS Comput Biol. 2013; 9(12): e1003365.
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