Fluorochromes for organelle tracing and multi-color imaging
Provided are compounds, methods and kits for identifying in cells of interest organelles including nuclei and a wide variety of organelles other than nuclei (non-nuclear organelles), as well as cell regions or cell domains. These compounds and methods can be used with other conventional detection reagents for identifying the location or position or quantity of organelles and even for distinguishing between organelles in cells of interest.
1. A compound comprising the structure
2. A method of identifying the location or position of a nucleus of a cell of interest, the method comprising the steps of:
(A) providing
(i) said cell of interest; and
(ii) the compound
(B) incubating said cell of interest with said compound; and
(C) identifying the location or position of said nucleus.
3. The method of claim 2 , further comprising the step of washing the cell after said incubating step and prior to said identifying step.
4. The method of claim 2 , further comprising the step of fixing said cell of interest prior to said incubating step.
5. The method of claim 2 , wherein said identifying step is carried out by detecting a fluorescent signal generated from said compound.
6. The method of claim 5 , wherein said fluorescent signal is detected by a means comprising a fluorescence microscope, a flow cytometer, a confocal microscope, a fluorometer, a microplate reader, a high-content cell analysis system, a high-content cell screening system, a laser-scanning cytometer, or a combination of any of the foregoing.
7. The method of claim 2 , further comprising providing one or more detection reagents which are different from said compound.
8. The method of claim 7 , wherein said one or more detection reagents are applied in said incubating step at the same time with said cell of interest and said compound.
9. The method of claim 7 , wherein said one or more detection reagents are applied to said cell of interest prior to said incubating step with said compound.
10. The method of claim 7 , wherein said one or more detection reagents are applied to said cell of interest after said incubating step with said compound.
11. The method of claim 7 , wherein said one or more detection reagents comprise an antibody, a lectin, an avidin, a streptavidin, a fluorescent protein, a fluorescent stain, a fusion tag, a phospholipid-binding protein, or a combination of any of the foregoing.
12. The method of claim 11 , wherein said one or more detection reagents comprise a fluorescent protein or fluorescent stain that comprises 4′,6-diamidino-2-phenylindole (DAPI), 5-Hydroxytryptamine (HAT), Acridine Orange, Acridine Yellow, Alexa Fluor dye, Blue Fluorescent Protein, BODIPY dye, Cascade Blue, Coelenterazine, Coumarin, Cyan Fluorescent Protein, Cyanine dye, Dansyl dye, Erythrosin, Far-red Fluorescent Proteins, FLUO 3, Fluorescein, FURA 2, Green Fluorescent Protein, Hoechst dye, INDO 1, JC-1 dye, Lucifer Yellow, Nile Red, Oregon Green dye, Propidium Iodide, QUIN 2, Red Fluorescent Protein, Rhodamine dye, R-Phycoerythrin, R-Phycoerythrin-Texas Red, SNARF, Texas Red, UV-excitable Green Fluorescent Protein, Yellow Fluorescent Protein, or a combination of any of the foregoing.
13. The method of claim 11 , wherein said one or more detection reagents comprises a fluorescent protein or fluorescent stain that comprises Cy5, Phyocoerythrin-Cy5 (PE-Cy5), Alexafluor 660, Alexafluor 680, a Spectrum FarRed conjugate, or a combination of any of the foregoing.
14. The method of claim 2 , wherein said cells of interest are contained in a sample.