Question: What Color Is PE Fluorescence?

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 is PE conjugated antibody?

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 PE flow cytometry?

In order to be used for flow cytometry, PE or APC must first be conjugated to an antibody. The antibody serves as the detector for the target of interest, while the PE or APC acts as the reporter which translates the detected event into a quantifiable signal.

What is multicolor flow cytometry?

Multicolor flow cytometry is a rapidly evolving technology that uses multiple fluorescent markers to identify and characterize cellular subpopulations of interest, allowing rapid analysis on tens of thousands of cells per second, with the possibility of isolating pure, viable populations by cell sorting for further …

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.

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.

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?

Most recent answer. In my experience, it is not advisable to use APC/Cy7 and PE/Cy7 together due to heavy cross-beam contamination as their emission max are exactly same. However, they can be used together if you are using machines like the Cytek Aurora.

What is the difference between FACS and flow cytometry?

FACS is used as a cell sorter and enriched for a subset of cells which is often then studied in further detail using flow cytometry or other analytical techniques2. Flow cytometry is used for cell analysis and is focused on measuring protein expression or co-expression within a mixed population of cells.

How do you compensate flow cytometry?

This is done by measuring the spectral overlap of the different fluorochromes and using the measured values to create a matrix. The matrix is then inverted and gives the actual compensation values. The flow cytometer then uses these values to correct the overlap in each detector for each colour.

How do you choose Fluorochromes for flow cytometry?

Fluorochromes are selected based on their abilities to fluoresce with the wavelengths of light produced by the lasers. Therefore, if a flow cytometer has only one laser that produces only 488 nm light, then only fluorochromes that are excited by 488 nm light can be used.

What is FACS analysis?

Fluorescence-activated cell sorting (FACS) is a specialized type of flow cytometry. It provides a method for sorting a heterogeneous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell.

What dyes can be used for flow cytometry?

Single dyes such as FITC, PE, APC and PerCP have been available for many years, but there are now alternatives available from Alexa Fluor dyes, which offer users greater photostability and brighter fluorescence.

What is the meaning of Phycobilins?

Phycobilins (from Greek: φύκος (phykos) meaning “alga”, and from Latin: bilis meaning “bile”) are light-capturing bilins found in cyanobacteria and in the chloroplasts of red algae, glaucophytes and some cryptomonads (though not in green algae and plants).

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.

What is the Colour of phycoerythrin?

redPhycoerythrin (PE) is a red protein pigment complex produced by the light-harvesting phycobiliprotein family. It is present in red algae and cryptophytes as an accessory to the main chlorophyll pigments responsible for photosynthesis (Ficner and Huber, 1993; van der Weij-De Wit et al., 2006).

Why is compensation of fluorescence necessary?

However, when emission spectra overlap, fluorescence from more than one fluorochrome may be detected. To correct for this spectral overlap, a process of fluorescence compensation is used. This ensures that the fluorescence detected in a particular detector derives from the fluorochrome that is being measured.

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.

What does red algae mean?

The red algae form a distinct group characterized by having eukaryotic cells without flagella and centrioles, chloroplasts that lack external endoplasmic reticulum and contain unstacked (stroma) thylakoids, and use phycobiliproteins as accessory pigments, which give them their red color.