Flow Cytometry Wavelength Chart
Bevezetés:
Flow cytometry is a powerful technique used in many fields of research, including immunology, cancer research, and stem cell studies. It allows researchers to analyze and quantify various characteristics of individual cells in a heterogeneous population. One crucial component of flow cytometry is the use of different wavelengths of light to excite and detect fluorochromes. In this article, we will discuss the flow cytometry wavelength chart, which outlines the commonly used fluorochromes and their respective excitation and emission wavelengths.
én. Fluorochromes and Excitation Wavelengths:
1. FITC (Fluorescein isothiocyanate):
– Excitation wavelength: 488 nm
– Emission wavelength: 530 nm
2. PE (Phycoerythrin):
– Excitation wavelength: 488 nm
– Emission wavelength: 578 nm
3. PerCP (Peridinin-chlorophyll-protein complex):
– Excitation wavelength: 488 nm
– Emission wavelength: 675 nm
4. BV421 (Brilliant Violet 421):
– Excitation wavelength: 405 nm
– Emission wavelength: 421 nm
II. Fluorochromes and Emission Wavelengths:
1. APC (Allophycocyanin):
– Excitation wavelength: 633 nm
– Emission wavelength: 660 nm
2. PE-Cy5 (Phycoerythrin-Cyanine5):
– Excitation wavelength: 488 nm, 633 nm
– Emission wavelength: 675 nm
3. Alexa Fluor 488:
– Excitation wavelength: 488 nm
– Emission wavelength: 519 nm
4. PerCP-Cy5.5 (Peridinin-chlorophyll-protein complex-Cyanine5.5):
– Excitation wavelength: 488 nm
– Emission wavelength: 695 nm
III. Application of the Flow Cytometry Wavelength Chart:
The flow cytometry wavelength chart serves as a reference tool for researchers involved in flow cytometry experiments. By knowing the excitation and emission wavelengths of different fluorochromes, scientists can design experiments to minimize the spectral overlap between fluorochromes and select appropriate filter settings for their flow cytometers. This ensures accurate and reliable data acquisition in flow cytometry analyses.
Moreover, the wavelength chart helps researchers in panel design. By considering the spectral properties of fluorochromes, they can select a combination of fluorochromes that minimize the fluorescence spillover into adjacent channels. This is particularly important when designing multi-color flow cytometry panels, where multiple fluorochromes are used simultaneously to analyze multiple cellular markers.
Következtetés:
The flow cytometry wavelength chart is an essential tool for researchers using flow cytometry in their studies. It provides information on the excitation and emission wavelengths of commonly used fluorochromes, enabling scientists to design experiments and optimize panel configurations for accurate and meaningful data acquisition. By utilizing the chart effectively, researchers can enhance the utility and reliability of flow cytometry as a scientific technique.
