Invitrogen™ Calcein AM, cell-permeant green and blue dyes

Description

• Calcein AM is a cell-permeant dye frequently used in cell biology research
• It is derived from Calcein, a fluorescent dye that exhibits bright green fluorescence upon binding to intracellular free calcium ions
• Calcein AM is a non-fluorescent compound that can permeate the cell membrane due to its lipophilic properties
• Once inside the cell, it is cleaved by intracellular esterases, yielding the fluorescent Calcein molecule
• This conversion allows the dye to be retained within the cytoplasm; apoptotic and dead cells with compromised membranes do not retain Calcein
• Cell vitality as measured by intracellular esterase activity is a recognized parameter of cell health
• Live cells are distinguished by the presence of ubiquitous intracellular esterase activity, determined by the enzymatic conversion of the virtually non-fluorescent cell permeant Calcein AM to the intensely fluorescent Calcein Green (ex/em 494/517 nm) or Calcein Blue (ex/em 360/449 nm)
• Apoptotic and dead cells with compromised membranes do not retain Calcein
• The acetoxymethyl (AM) ester derivatives of fluorescent indicators make up one of the most useful groups of compounds for the study of live cells
• Once inside the cell, the lipophilic blocking groups are cleaved by nonspecific esterases, resulting in a charged form that is retained in cells to a much greater extent than its parent compound
• In addition to cell viability assays, Calcein AM is also used to study cell migration, cell adhesion, and cellular uptake processes
• Its fluorescence properties make it a versatile tool for various applications in cell biology research
• Application of the non-AM Calcein cell-impermeant dye: Calcein, AM requires esterase cleavage of the acetoxymethyl (AM) ester to become fluorescent
• Liposomes don't have esterases unless specifically constructed to include the enzyme
• The water-soluble, non-AM form of Calcein (Cat
• No
• C481), does not require esterase cleavage to be fluorescent
• This versatile green fluorophore is cell impermeant and is commonly used for cell tracing and in studies of endocytosis and gap junctions
• Features of Calcein AM cell-permeant dye: Cell-permeant —Calcein AM is cell-permeant, meaning it can pass through the cell membrane and enter live cells
• This property allows researchers to label and visualize intracellular structures and processes
• Non-toxic —Calcein AM is non-toxic to cells and does not significantly affect cell function or metabolism
• Esterase cleavage —Once inside the cell, the acetoxymethyl (AM) ester group of Calcein AM is cleaved by intracellular esterases, resulting in the formation of Calcein
• This cleavage process activates the fluorescence of Calcein, allowing for easy detection and visualization
• Uniform distribution —Upon entering live cells, Calcein AM distributes uniformly throughout the cytoplasm and nucleus
• This even distribution allows for accurate assessment and visualization of cellular processes
• Stability —Calcein AM is relatively stable within cells and can be used for long-term imaging experiments without significant photobleaching
• Fluorogenic —Calcein AM is a non-fluorescent compound, but once inside the cell, it is cleaved by intracellular esterases, resulting in the formation of fluorescent Calcein
• Bright and Stable Fluorescence —Calcein exhibits bright green or blue fluorescence when it binds to intracellular free calcium ions or other targets
• The resulting fluorescence is stable, allowing for long-term imaging and analysis
• Application of the Calcein AM cell-permeant dye: Calcein AM can be used in various applications, including cell viability assays, cell migration and invasion studies, intracellular calcium imaging, cell labeling and tracing, and fluorescence microscopy and flow cytometry
• Cell viability —Calcein AM is frequently used to assess cell viability and cytotoxicity
• Live cells with intact cell membranes can take up Calcein AM, which is then cleaved by intracellular esterases to produce fluorescent Calcein
• The fluorescence intensity of Calcein can be measured to determine cell viability
• Cell migration —Calcein AM can be used to track and visualize cell migration
• By labeling cells with Calcein AM, their movement and migration can be monitored over time using fluorescence microscopy
• This is particularly useful in studying processes such as wound healing, metastasis, and cell trafficking
• Cell adhesion —Calcein AM can be used to study cell adhesion and cell-substrate interactions
• By labeling cells with Calcein AM and culturing them on various substrates, the adhesion properties of cells can be assessed
• Changes in fluorescence intensity can indicate differences in cell adhesion
• Cellular uptake —Calcein AM can be used to investigate cellular uptake processes
• By labeling molecules or nanoparticles with Calcein AM, their uptake by cells can be tracked and analyzed
• This is valuable in studying drug delivery mechanisms and understanding cellular uptake pathways
• Intracellular pH measurements —Calcein AM has been used as a pH-sensitive dye to measure changes in intracellular pH
• The fluorescence properties of Calcein can be modulated by pH, allowing researchers to monitor pH changes within cells
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