Cell Imaging Probes

Product#: BIOACT3
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Cell Imaging Probes

The live cell imaging with fluorescent probes can give opportunities to analyze cellular events in real–time thereby lead to new biological discoveries those unachievable with traditional analysis methods. Conventional techniques, such as flow cytometry and antibody immunocytochemistry staining, analyze cellular events as a snapshot rather than in real–time. We offer novel fluorescent cell imaging probes for cell structure analysis, apoptosis and angiogenesis imaging, ROS detection, etc.
 

Cell Structure Analysis Probes

FSD Fluor™ Phalloidin Series

Phalloidin is a rigid bicyclic peptide toxin isolated from Amanita phalloides mushroom that commonly used in imaging applications to selective label F-actin. Phalloidin is known to react stoichiometrically with actin, strongly promote actin polymerization, and stabilize actin polymers. Due to the ability to bind F-actin selectively, fluorescence conjugated phalloidin derivatives are widely used in microscopy for investigating the distribution of F-actin in cells. In biomedical research, fluorescent phalloidin probes are utilized in localizing actin filaments in living or fixed cells as well as for visualizing individual actin filaments in vitro. Fluorescent phalloidins much smaller than fluorescent antibodies, thus they have several advantages for actin labeling such as virtually identical binding properties with actin from different species of plants and animals, much denser labeling of filamentous actin, more detailed images, and lower nonspecific binding. Fluorescent phalloidin conjugates are not permeable to most live cells, thus they should be used to detect cells with compromised membranes. However, fluorescent labeled phalloidins may penetrate the membranes of certain hypoxic cells. BioActs offers FSD Fluor™ Phalloidin series for the labeling and quantitative analysis of F-actin in formaldehyde-fixed and permeabilized tissue sections, cell cultures, and cell-free experiments.

CytoFlamma® Cell Membrane

Lipophilic organic dyes bear some structural resemblance to natural lipids and can be incorporated into cell wall and liposome. BioActs developed CytoFlamma® cell membrane series as versatile membrane selective fluorescent probes. The probes consist of Flamma dye attached with hydrophobic lipid analogs. They can be used in studies for biophysical analysis of membranes, for tracing lipid transport and metabolism in live cells, and for lipid-mediated signal transduction processes. Flamma® dyes display strong absorption, high fluorescence quantum yield and high photostability, and they maintain good fluorescence activity and stability after conjugation to biomolecules. Due to low toxicity and stable retention, CytoFlamma® cell membrane probes might be useful for long-term cell tracing and can be used as membrane markers of endocytosis and exocytosis. The probes are stable under photooxidation effects induced by ultraviolet excitation and are resistant to spontaneous oxidation. We offer CytoFlamma® cell membrane series as selective fluorescent probes for analyzing cell structure by incorporating into plasma membrane.

MitoFlamma® Green

MitoFlamma® Green is a mitochondria-selective green fluorescent dye that allows to detect mitochondrial morphology in living cells. This unique dye appears to preferentially accumulate in mitochondria regardless of mitochondrial membrane potential in certain cell types, making it a possible tool for determining mitochondrial mass. MitoFlamma® Green enables researchers to observe mitochondrial activity, localization and abundance as well as monitoring the effect of drugs or other external stimuli on the mitochondrial function. The maxima of excitation/emission are at 508/580 nm. Once mitochondria are labeled with this special dye, the cells can be treated with an aldehyde fixative for samples that require fixation for further processing. MitoFlamma® Green probes are retained in the mitochondria during the fixation step and after permeabilization with some detergents during subsequent processing steps. After fixation, labeled samples can be applied in a variety of experiments such as immunocytochemistry, in situ hybridization, microplate-based analysis, etc.

Cytoskeleton

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Plasma Membrane

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Mitochondria

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Apoptosis Imaging Probes


Annexin V Flamma® series

In normal live cells, phosphatidyl serine (PS) is located on the cytoplasmic surface of the cell membrane, however PS is translocated to the outer leaflet of the plasma membrane exposing to the external cellular environment when apoptosis is in progress. Annexin V is a Ca2+ dependent PS-binding protein that has been used as a non-quantitative probe to detect cells that have expressed PS on the cell surface. Fluorescence conjugated annexin V enables to detect the externalized PS by binding to PS that exposed on the outer leaflet. BioActs developed a series of Flamma® dyes and other dyes such as FITC, TAMRA and ICG conjugated annexin V, Annexin V-Flamma® series, as PS selective fluorescent probes.
Annexin V Flamma® series is a rapid and selective probe for labeling of externalized PS, an indicator of intermediate stages of apoptosis. Flamma® dyes display strong absorption, high fluorescence quantum yield and high photostability, and they maintain good fluorescence activity and stability after conjugation to biomolecules, allowing the detection of low-abundance biological structures with great sensitivity. Annexin V Flamma® conjugates can be utilized in fluorescent imaging as well as for flow cytometry of apoptotic cells, and they might be used in combination with other dyes such as propidium iodide (PI) in order to accurately analyze mixed populations of apoptotic and other cells. Annexin V Flamma® probes available as stand-alone reagent or easy-to-us kits.
Annexin V Flamma® apoptosis detection kit includes a PI nucleic acid binding dye and annexin V binding buffer. PI is impermeant to live cells and apoptotic cells, but stains dead cells with red fluorescence by binding to nucleic acids. Normal live cell is not labeled by ether Annexin V or PI, early stage apoptotic cell is stained with Annexin V and natural death or cell subject to necrosis is stained with PI. The late stage apoptotic cells are stained by both reagents. After staining a cell population with Annexin V Flamma® and PI in annexin binding buffer, apoptotic cells and dead cells can be distinguished and analyzed using a flow cytometer.

Flamma® Fluors TUNEL Assay Kit

Apoptosis is a highly regulated and controlled process that confers advantages during an organism's lifecycle, and defective apoptotic processes have been implicated in a variety of diseases such as atrophy and cancers. In situ labeling of apoptotic cells allows highly sensitive and quantitative analysis of involved cells. Characteristics of the late stages of apoptosis are changes in nuclear morphology, including chromatin condensation, degradation of nuclear envelope, and DNA strand breaks. TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling) assays are the most widely used methods for rapid identification and quantification of the apoptotic cell fraction in cultured cell preparations. TUNEL reaction preferentially labels DNA strands that generated during apoptosis using the ability of TdT to label blunt ends of double-stranded DNA independent of a template. This selective labeling allows discrimination of apoptosis from necrosis and from primary DNA strand breaks induced by cytostatic drugs or irradiation.
BioActs developed Flamma® Fluors TUNEL assay kit for quantitative analysis of apoptotic cells in the mixed cell populations. The kit employs dUTP conjugated Flamma® Fluors and FITC dyes for labeling of fixed cells or tissues. Flamma® Fluor dyes display excellent fluorescence activity and photostability after conjugated to biomolecules, allowing detection of low-abundance biological structures with great sensitivity. Flamma® TUNEL assay kit is reliable and effective method to detect and quantify a wide levels of apoptotic cells by fluorescence microscopy and flow cytometry. In addition, the kit can be used in the analysis of apoptotic cell of paraffin-embedded tissue sections along with cultured cells. BioActs offers Flamma® TUNEL assay kit for the detection apoptotic cells in frozen and formalin-fixed tissue sections, determination of the sensitivity of malignant cells to drug-induced apoptosis, discrimination of apoptotic and necrotic cells in the cell death environment, etc.

 

Annexin V Series

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TUNEL Assay Kit

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ROS Detection Probes

Hydrogen peroxide (H2O2) plays important roles in various cellular signal transductions in physiological processes as well as pathological effects. It has shown that H2O2 is the main reactive oxygen species (ROS) in redox signaling during cell proliferation and apoptotic process. Thus, quantitative analysis of endogenous H2O2 might to be a useful tool for understanding cell physiology. BioActs developed a ROS reactive fluorescence probe NpFlamma® ROS 380 as an efficient detecting tool for fast, sensitive and selective imaging of low level H2O2 generated during various cellular processes.

NpFlamma® ROS 380 comprised of a physical assembly of H2O2 reactive fluorophore and catalytic additive into an amphiphilic stabilization colloid. This versatile reactive nanoparticle is hardly fluorescent in inactivated state yet displays a strong red-shifted emission upon oxidized by H2O2. The maxima of Excitation/emission are at 380/450 nm. Since the emission is achieved by a chemical reaction, the interference between background fluorescence and the excitation light irradiation cannot occur. NpFlamma® ROS 380 allows a real-time imaging of low-level H2O2 involved in cellular processes along with oxidative stress. We offer NpFlamma® ROS 380 as a quantitative H2O2 selective fluorescence probe for biological and physiological analysis involving oxidative cell processes.

NpFlamma® ROS

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Angiogenesis Probes

Integrins are heterodimeric transmembrane receptors for cell adhesion to extracellular matrix (ECM) proteins and play important roles in certain cell-cell adhesions. In addition, they make transmembrane connections to the cytoskeleton and activate many intracellular signaling pathways. Because of their role in tumor angiogenesis and progression, integrins have become important diagnostic and therapeutic targets. To date 24 distinct integrins are known, and among them, integrin αvβ3 is known to strongly involve in the regulation of angiogenesis. The αvβ3 is generally expressed in low levels on the epithelial cells and mature endothelial cells, but it is highly expressed in many solid tumors. The αvβ3 levels correlate well with the potential for tumor metastasis and aggressiveness, which make it an important biological target for development of antiangiogenic drugs and molecular imaging probes for early tumor diagnosis.

Cyclic tripeptide arginine-glycine-aspartate (RGD) is well-known to bind preferentially to αvβintegrin with high affinity. Cyclic RGD is an effective ligand for tumor targeting since integrin αvβis overexpressed not only on tumoral endothelium but also on various cancer cells. Thus, targeting tumor cells or tumor vasculature by RGD-based strategies is a promising approach for delivering anticancer drugs or diagnostic agents. Cyclic RGD peptides are also able to bind αvβ5, α5β1, α6β4, α4β1, and αvβ6 integrins, which may help enhance their tumor uptake due to the increased receptor population. BioActs developed AngioFlamma® series as effective fluorescent probes for the detection of angiogenesis and tumor cells. The probes are made up of various fluorophores conjugated cyclic RGD. We offer AngioFlamma® series as in vivo fluorescent probes for imaging of blood vessels, tumors and angiogenesis.

AngioFlamma® Series

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