It was found that fluorescence intensity of Cy5-, Cy7-Texas Red- and Dylight 649-NeutrAvidin conjugates as well as TMR-dextran was generally insensitive to pH over a range of 3 to 10.
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Is Cy3 pH sensitive?
Cy3 excites maximally at 544 nm and has an emission peak at 564 nm (Figure 3B), and produces a pH-insensitive synchronous scan peak at 548 nm (Figure 4).
Is fluorescein pH sensitive?
One of the most notorious pH-sensitive probes is fluorescein. In addition to excellent photophysical properties, this fluorophore presents a pH-sensitivity around neutral and physiologic domains.
Is FITC light sensitive?
FITC is diluted in basic buffer for coupling procedures immediately prior to use. The products are light-sensitive, and should be stored dry and in the dark at 2 °C to 8 °C.
What does Cy3 stain for?
The Cy3 dye is a traditional orange-fluorescent label for protein and nucleic acid conjugates used for imaging, flow cytometry, and genomic applications. It is also the basis for the classic lipophilic tracer DiI and its variants.
What is Cy3 and Cy5?
Cyanine Cy3 and Cy5 are fluorescent dyes with many applications in nonlinear optics, laser physics, and particularly in biomedical imaging and single-molecule spectroscopy. Both dyes adopt an all-trans configuration in its ground state (Figure 1) and can be easily attached to nucleic acids and proteins.
What Colour is FITC?
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.
Is FITC pH sensitive?
FITC conjugates are not very stable to elevated temperatures and are very pH sensitive (in terms of their fluoresence behavior; fluoresence yield was found to be higher at pH 7.4 than at pH 6.0 by John N.
How does pH affect fluorescein?
The fluorescence of Fluorescein increased 1.3 times, Tinopal 1.25 times, and Pyranine 3.0 times as the pH value increased from 6.9– 8.4, but it became nearly constant when pH value was greater than 8.4.
What is the effect of pH on fluorescence?
In all samples, fluorescence excitation brightness values increased as pH increased, whilst peak fluorescence decreased with increasing culture age at equivalent pH. Excitation emission matrices of suspensions of P. intermedia after 24 hours growth. Adjusted to acidic (5.74), neutral (7.14) and alkaline (8.63) pHs.
Is Alexa Fluor 488 same as FITC?
Alexa Fluor 488 (Ex-Max 495 nm/Em-Max 519 nm): have nearly identical emission and excitation maxima as FITC. However, Alexa Flour 488 display higher photostability and is less susceptible to self-quenching.
Is fluorescein same as FITC?
Fluorescein derivatives are the most common fluorescent reagents for biological research because of their high absorptivity, excellent fluorescence quantum yield, and good water solubility.
What is DAPI staining used for?
DAPI staining was used to determine the number of nuclei and to assess gross cell morphology. Following light microscopic analyses, the stained cells were processed for electron microscopy. Cells stained with DAPI showed no ultrastructural changes compared to the appearance of cells not stained with DAPI.
What does cyanine mean?
Definition of cyanine : any of various dyes used especially to sensitize photographic film to light from the green, yellow, red, and infrared regions of the spectrum.
Which of the fluorescent Colour light is generally used in microarray experiments?
The microarray is analyzed in a scanner, which uses laser light to cause each spot to fluoresce and measures the relative intensity of emission, typically at green (Cy3) and red (Cy5) wavelengths.
What Colour is Cy5?
Cy5 dye is a traditional far-red–fluorescent label for protein and nucleic acid conjugates used for imaging, flow cytometry, and genomic applications.
Can you use Cy3 and Cy5 together?
Cy3 dyes are yellow-orange. Specific filter wavelengths (550-600 nm and 655-695 nm) have to be used to avoid background contamination when using Cy3 and Cy5 together to optically separate the two fluorescent cyanines. Cy3 is used interchangeably with PE, though PE remains a better choice.
What wavelength is FITC?
FITC emits fluorescence from 475 to 650 nm, peaking at 525 nm, which falls in the green spectrum.
What color is Cy7?
Cy7 is a near-IR fluor that is invisible to the naked eye.
What is fluorescein dye used for?
Fluorescein is a diagnostic contrast agent particularly used in various ophthalmic procedures, such as checking for any corneal or vessel abnormalities. The application of fluorescein also extends to bioimaging whole anatomic structures and even further to cellular components in immunohistological staining.
What is FITC in biology?
Fluorescein isothiocyanate (FITC) is the most widely used fluorescent probe for the preparation of conjugates with biological molecules (Hansen, 1967; Haugland, 1990).
What wavelength is DAPI?
Fluorescence properties When bound to double-stranded DNA, DAPI has an absorption maximum at a wavelength of 358 nm (ultraviolet) and its emission maximum is at 461 nm (blue). Therefore, for fluorescence microscopy, DAPI is excited with ultraviolet light and is detected through a blue/cyan filter.
Is FITC a fluorescent dye?
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 is the pH of fluorescein?
The strong fluorescein dianion emission (max. at 515 nm) appears in alkaline solution. In the pH range of existence of the fluorescein monoanion (at about pH 5.5), the fluorescence spectrum isessentially the same as that of the neutral molecule (with a slight contribution of dianion emission).
Can I use FITC and PE together?
FITC overlap a lot with Pe, don’t use them together on same cells. PerCP-‐Cy5. 5 overlap surprisingly a lot with BV711, don’t use together.