What Is PE Flow Cytometry?

What color is PE fluorescence?

R-phycoerythrin (R-PE) is an intensely bright phycobiliprotein isolated from red algae that exhibits extremely bright red-orange fluorescence with high quantum yields.

It is excited by laser lines from 488 to 561 nm, with absorbance maxima at 496, 546, and 565 nm and a fluorescence emission peak at 578 nm..

What is FITC in flow cytometry?

Fluorescein isothiocyanate (FITC) is a derivative of fluorescein used in wide-ranging applications including flow cytometry. … FITC has excitation and emission spectrum peak wavelengths of approximately 495 nm/519 nm, giving it a green color. Like most fluorochromes, it is prone to photobleaching.

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.

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.

What is the principle of flow cytometry?

The basic principle of flow cytometry is the passage of cells in single file in front of a laser so they can be detected, counted and sorted. Cell components are fluorescently labelled and then excited by the laser to emit light at varying wavelengths.

What is the difference between FACS and flow cytometry?

The terms flow cytometry and fluorescence-activated cell sorting (FACS) are often used interchangeably. … FACS is a derivative of flow cytometry that adds an exceptional degree of functionality. Using FACS a researcher can physically sort a heterogeneous mixture of cells into different populations.

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 PE stand for in flow cytometry?

Phycoerythrin (PE) is one of the most commonly-used fluorescent dyes for FACS analysis. PE is a large protein (approximate molecular weight 240 kd) containing 25 fluors. Typically, only one PE molecule is conjugated to an antibody.

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™.

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.

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 is APC in flow cytometry?

APC is a large molecule that is chemically crosslinked to use in fluorescence-based detection, primarily for flow cytometry, microarray assays, ELISAs, and other applications that require high sensitivity but not photostability.

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.

How do you pay for flow cytometry?

How To Compensate A 4-Color Flow Cytometry Experiment Correctly4 Steps To Compensating A 4-Color Experiment. … Choose the correct carrier for compensation. … Step 2: Collect the data and make sure there is a sufficient number of events. … Calculate compensation correctly. … Apply the compensation values and inspect the results.

What is PE conjugated antibody?

Overview: Phycoerythrin (PE) is one of the most commonly-used fluorescent dyes for FACS analysis. PE is a large protein (approximate molecular weight 240 kd) containing 25 fluors. Typically, only one PE molecule is conjugated to an antibody. … (See the fluorescence spectra and the absorbance spectrum).

What color is FITC?

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

What is R in R phycoerythrin?

R-phycoerythrin (R-PE) is an intensely bright phycobiliprotein isolated from red algae that exhibits extremely bright red-orange fluorescence with high quantum yields. It is excited by laser lines from 488 to 561 nm, with absorbance maxima at 496, 546, and 565 nm and a fluorescence emission peak at 578 nm.