Can I Use APC Cy7 And PE Cy7 Together?

What is PE flow cytometry?

R-PE is a large molecule used for fluorescence-based detection, primarily in flow cytometry, microarray assays, ELISAs, and other applications that require high sensitivity but not photostability.

Long-wavelength fluorescence emission..

What color is PE?

488 nm 532 nm 561 nm 575/26 PE (Exmax 496 nm/Emmax 578 nm) R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae.

Can flow cytometry detect dead cells?

Loss of membrane integrity is a definitive indicator of cell death in flow cytometric assays. Cells that exclude a dead cell dye are considered viable, while cells with a compromised membrane allow the dye inside into cell to stain an internal component, thus identifying the cell as dead.

Why flow compensation is required?

Mass flow of steam is measured indirectly by measuring the differential pressure acting across an orifice plate. The measurement requires compensation because it is affected by the temperature and pressure existing at the measurement time, and the result is not linear relative to the differential pressure.

What dyes can be used for flow cytometry?

Table 2. Tandem dyes for flow cytometryFluorophoresFluorescence ColorRelative BrightnessPE-Cy5.54PE-Alexa Fluor® 750Infrared4PE-Cy7Infrared2APC-Cy723 more rows

What does FITC bind to?

Fluorescein isothiocyanate (FITC) is widely used to attach a fluorescent label to proteins vi a the amine group. The isothiocyanate group reacts with amino terminal and primary amines in proteins. It has been used for the labeling of proteins including antibodies and lectins. 2-6.

How does compensation work in flow cytometry?

In cytometry, compensation is a mathematical correction of a signal overlap between the channels of the emission spectra of different fluorochromes. … The compensation can be done through different flow cytometry software such as Flowjo, Flowlogic, Kaluza etc.

Can flow cytometry detect leukemia?

Flow cytometry is an indispensable tool for diagnosis and monitoring of leukemia and lymphoma. While application of flow cytometry in this field may be complex and require a lot of experience, it is based on rather simple principles.

What does FITC stain?

Fluorescein-5-isothiocyanate (FITC) is a fluorescence dye and belongs to the xanthene dyes. FITC is used for labeling of different biomolecules, e.g. immunoglobulins, lectins and other proteins, peptides, nucleic acids, nucleotides; oligo-and polysaccha- rides.

Can I use PE and PE cy7 together?

When using tandem antibody conjugates in multicolor staining panels, it is important to use exactly the same tandem conjugate for compensation tubes that are used for staining experiment samples. … PE-Cy7 (BD) ≠ PE-Cy7 (Biolegend) for any antibody conjugate. CD4 PE-Cy7 (BD) ≠ CD8 PE-Cy7 (BD) for compensation.

What is the most common clinical application of flow cytometry?

The most common application performed on the cytometer is immunophenotyping. This technique identifies and quantifies populations of cells in a heterogeneous sample – usually blood, bone marrow or lymph.

What color is FITC?

green colorFITC has excitation and emission spectrum peak wavelengths of approximately 495 nm/519 nm, giving it a green color.

How do you choose Fluorochromes for flow cytometry?

Rule 1: Choose the brightest set of fluorochromes for your particular instrument configuration. Rule 2: Choose fluorochromes to minimize the potential for spectral overlap. Rule 3: Reserve the brightest fluorochromes for dim antibodies and vice versa.

What color is APC?

Allophycocyanin (APC) Allophycocyanin (APC) is an intensely bright phycobiliprotein isolated from red algae that exhibits far-red fluorescence with high quantum yields. It is excited by laser lines at 594 and 633 nm, with an absorbance maximum at 650 nm and a fluorescence emission peak at 660 nm.

How does flow cytometry work?

The flow cytometer: fluidics The tiny stream of fluid takes the cells past the laser light one cell at a time (Figure 1). Light scattered from the cells or particles is detected as they go through the laser beam. … Fluorescence detectors measure the fluorescence emitted from positively stained cells or particles.

Why use isotype controls flow cytometry?

Isotype controls are primary antibodies that lack specificity to the target, but match the class and type of the primary antibody used in the application. Isotype controls are used as negative controls to help differentiate non-specific background signal from specific antibody signal.

What is FITC and PE?

The FITC / PE Compensation Standard is to be used in conjunction with hardware or software to remove spectral overlap from fluorochromes into secondary fluorescence detectors of a flow cytometer. … The FITC/PE Compensation Standard is a mixture of 4 populations of microspheres: FITC, PE, FITC/PE, and AutoFluor™.

Why is phycoerythrin red?

Red algae are red because of the presence of the pigment phycoerythrin; this pigment reflects red light and absorbs blue light. … Some rhodophytes have very little phycoerythrin, and may appear green or bluish from the chlorophyll and other pigments present in them.