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.
What is Allophycocyanin used for?
Allophycocyanin, also known as APC, is typically used for flow cytometry, microarrays, ELISA, and other applications that require high sensitivity but not photostability.
What is APC channel 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.
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 Phycocyanin and Allophycocyanin?
Phycocyanin is a pigment-protein complex from the light-harvesting phycobiliprotein family, along with allophycocyanin and phycoerythrin. It is an accessory pigment to chlorophyll. … In addition, fluorescence detection of phycocyanin pigments in water samples is a useful method to monitor cyanobacteria biomass.
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.
What is PE conjugated antibody?
Phycoerythrin (PE) is a type of phycobiliproteins found in cyanobacteria and red algae. PE-conjugated antibodies are broadly used for flow cytometry and immunofluorescence microscopy. … However, nonspecific binding of specific cell types by a fluorescent dye-conjugated form of antibody has been rarely reported.
Where is Phycocyanin found?
C-phycocyanin is a protein-bound pigment soluble in water and found in some blue-green microalgae such as Spirulina, which is used in many countries as a dietary supplement. According to Romay et al. (1998b), phycocyanin shows anti-inflammatory activity in four experimental models of inflammation.
What laser is APC?
APC is excited by the red diode laser and excites in several tandem dyes including APC-Cy5. 5 and APC-Cy7. Alexa Fluor 647 (Ex-Max 650 nm/Em-Max 668 nm): conjugates are highly photostable and remain fluorescent over a broad pH range.
Does PE and FITC overlap?
In some experiments FITC may be combined with other dyes, for example PE, that emit yellow and orange photons. Fig. … FITC spillover into other channels. FITC single stained lymphocytes show spillover into PE and PE-Cy5 detectors.
Does PE and APC overlap?
Furthermore, PE (and PE-Texas Red to an even greater extent) does overlap significantly with APC.
Can I use APC and PE together?
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.
Which fluorophores are brightest?
It is concluded that cyanine 3.18 should be useful for multicolor fluorescence experiments and that it may be the brightest fluorophore available for single-color fluorescence immunocytochemistry.
Is Alexa Fluor 700 a tandem?
Additionally, we have conjugated allophycocyanin to Alexa Fluor 680, Alexa Fluor 700 or Alexa Fluor 750 dyes to create tandem conjugates for multicolor measurements using excitation sources in the 633 to 650 nm range (Figure 1.3. 25).
What is fluorescent chemical?
Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation. It is a form of luminescence. … Fluorescent materials cease to glow nearly immediately when the radiation source stops, unlike phosphorescent materials, which continue to emit light for some time after.
What is spirulina do?
Spirulina is a type of cyanobacteria — often referred to as blue-green algae — that is incredibly healthy. It may improve your levels of blood lipids, suppress oxidation, reduce blood pressure and lower blood sugar.
Are there any side effects to taking spirulina?
Some minor side effects of spirulina may include nausea, insomnia, and headaches. Still, this supplement is widely considered safe, and most people experience no side effects (2). Spirulina may be contaminated with harmful compounds, thin your blood, and worsen autoimmune conditions.
How do Fluorochromes work?
Fluorochromes absorb light energy of a specific wavelength and re-emit it at a longer wavelength. … You can find commercially available fluorescent dyes that absorb and emit light at specific wavelengths across the whole visible spectrum, including the infrared region.
Are PE and R PE the same?
* PE is the same as R-phycoerythrin. APC, allophycocyanin; PE, phycoerythrin; PerCP, peridinin chlorophyll protein.
What color is PE?
Field | Color |
---|---|
Physical Education | Sage green |
Public Admin. including Foreign Service | Peacock blue |
Public Health | Salmon pink |
Science | Golden yellow |
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 is FITC antibody?
FITC (fluorescein isothiocyanate) is a fluorochrome dye widely used as an antibody or other probe marker. FITC absorbs ultraviolet or blue light, exciting molecules which then emit a visible yellow-green light. When the excitation light is removed, the emission light ceases.
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 and 519 nm, giving it a green color. Like most fluorochromes, it is prone to photobleaching.
What are the benefits of phycocyanin?
Phycocyanin can promote animal blood cell regeneration, improve lymphocyte activity and lymphatic system to improve immune function, and comprehensively enhance the disease resistance of the body 73.
What is phycocyanin spirulina?
Phycocyanin is a pigment-protein complex synthesized by blue-green microalgae such as Arthrospira (Spirulina) platensis. This pigment is used mainly as natural colouring in food industry. … The phycocyanin extraction was performed using water at the different biomass-to-solvent ratio and shaking rates.
Is phycocyanin a carotenoid?
Arthrospira platensis is the widely available source of spirulina that contains distinctive natural pigments, including carotenoids and C-phycocyanin (C-PC). … The carotenoid profiles suggested that the commercial spirulina powders originated from two different sources, A. platensis and A.
What laser is PE on?
PE is excitable by both the 488 nm laser and 561 nm laser, therefore PE and its tandem dyes appear here and on the 488 nm excitable fluorophores web page. As PE is maximally excited at 546 nm (compared to 496 nm using the 488 nm laser), where possible it is advisable to use the 561 nm laser to get the maximal signal.
Is APC brighter than PE?
Compared to PE, APC is not as”bright”. … Compared to other red fluorophores such as Cy5, however, APC is still significantly brighter and an excellent choice for use in flow cytometry.
What laser does PE use?
The blue laser is versatile due to its ability to excite both FITC (and similar fluorophores), PE, and PE tandems. PE is also excitable by the 561 nm laser and so PE based fluorophores appear on both lists.
Is af488 same as FITC?
Molecular Probes™ Alexa Fluor® 488 dye—with nearly identical spectral properties and quantum yield as fluorescein (FITC)—produces brighter, more photostable conjugates that are ideal for imaging and other applications requiring increased sensitivity and environmentally insensitive fluorescence detection.
Does FITC bleed into PE?
where FITC bleeds into the PE channel and PE bleeds back into FITC. To correct for spectral overlap during multicolor flow cytometry experiments, color compensation must be performed. The goal of color compensation is to correctly quantify each dye with which a particular cell is labeled.
Are FITC and GFP the same?
Green Fluorescent Protein (GFP) Antibody (FITC) is a Rabbit Polyclonal antibody conjugated to FITC against GFP.
How do you compensate FACS?
- 4 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.
Is BV421 same as Pacific Blue?
BV421™ has an emission spectrum that is more narrow than that of Pacific Blue™, thus, there is less spillover into neighboring channels, such as AmCyan or Horizon™ V500.
Is BV510 a tandem?
BD Horizon Brilliant™ Violet 510 (BV510) (Exmax 405 nm/Emmax 510 nm) is a polymer-based dye that is brighter than BD Horizon™ V500. … 405 nm 610/20 BD Horizon Brilliant™ Violet 605 (BV605) (Exmax 407 nm/Emmax 602 nm) is a tandem fluorochrome that combines BD Horizon BV421 and an acceptor dye with emission at 602 nm.
What Colour is FITC?
FITC exhibits an excitation maximum at λ = 495 nm and emission maximum at approximately λ = 519 nm. The color of the compound is yellow while the emitted light is green.
What is pecy7?
PE-Cy7 is a tandem fluorochrome consisting of R-phyco- erythrin (PE), a protein which is excited by an Argon laser at 488 nm and acts as an energy donor, coupled to the cyanine dye Cy7, a small organic molecule which is an energy acceptor and emits at 767 nm.
What is BV421?
BD Horizon™ Brilliant Violet™ 421 (BV421) (Exmax 407 nm/Emmax 421 nm) is a polymer-based dye excited by the violet laser and is one of the brightest fluorochromes offered by BD Biosciences. … This dye offers improved brightness over Pacific Orange™ and reduced spillover into the FITC channel when compared to AmCyan.
What is the difference between fluorophore and fluorochrome?
As nouns the difference between fluorochrome and fluorophore
is that fluorochrome is any of various fluorescent dyes used to stain biological material before microscopic examination while fluorophore is (biochemistry) a molecule or functional group which is capable of fluorescence.
What determines fluorophore brightness?
The brightness of a fluorescent molecule depends on two factors: Extinction Coefficient (ε) – how strongly a fluorescent molecule absorbs light at a particular wavelength. Fluorescence Quantum Yield (Φ) – how efficiently the absorbed light is converted into emitted light.
What color is Alexa Fluor 594?
Invitrogen Alexa Fluor 594 dye is a bright, red-fluorescent dye that can be excited using the 561 nm or 594 nm laser lines.