BioLegend's Fluorescence Spectra Analyzer is useful for the analysis of excitation and emission spectra of commonly used fluorochromes for flow cytometry.
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PE/Fire™ 780 is an equivalent fluor to PE/Cy7. It is restricted to ASR use only and is only available to US customers.
Some fluorochromes (e.g. Spark Blue™ 550, Spark NIR™ 685) are only recommended for use in multicolor spectral flow cytometry panels. To view the spectral signatures of fluorochromes in a spectral flow cytometer, please visit the Aurora Spectra Analyzer.
|1. View Spectra|
|To view spectra, simply select a fluorophore from one of the dropdowns, then select or deselect the emission and/or excitation spectrum of your fluorochrome of interest. The graph legend appears above the display, indicating the displayed spectrum. Excitation spectra are displayed as dotted lines, while emission spectra are displayed as smooth lines.|
|2. Select Laser Lines|
|To visualize laser lines select your desired line from the listed options|
|3. Create Filters|
|If you wish to visualize the degree of fluorescence transmission through your filter sets, enter the filter wavelength and bandpass into column 4. Turn your filters on and off using the checkboxes.|
|4. Pinpoint spectral data|
|To view exact data points on any graph, make sure the "Enable tooltip" box is selected and then mouse over spectra curves until a popup appears. The popup displays the fluorochrome, excitation or emission, the wavelength of light, and the % of maximum excitation or emission at that wavelength. For iPhone and iPad users, simply press on the location of the curve to view the same data display.You can also pinpoint your mouse position using the Mouse Position indicator below the selection area. Note that this tool locates the position of the mouse regardless of the spectra. It is useful for pinpointing data points that may be difficult to find using the tooltip popup box.|
|5. Zoom in, zoom out, and pan|
|To help with pinpointing exact data, the analyzer has the capacity to zoom in, allowing you easily visualize spectra intersection points with laser lines, filter sets and other spectra. To zoom in, double click your mouse over the area that you wish to zoom, or roll the mouse wheel forward. To zoom out, click on the zoom out button at the top right corner or roll the mouse wheel backward. To pan, click anywhere within the graph and move the cursor to your desired position. To reset the data to the original layout, click on the Reset Zoom button.|
|6. Customize Colors|
|All the colors for spectra, laser lines, and filters are customizable. After you have made your selections, click the Customize Colors button to open the interface panel. To change colors, click on the desired color box. Select colors by dragging the locator within the box gradient and/or use the color slider. You can also enter in RGB values if known. To go back to the default color, click on the top right box which shows the default color.|
|7. Export Image|
|To export and save the data as an image, click the Export Spectra button. A new window with the data will appear as a png file. Save it by right clicking and select “Save image as” or select “Copy image” to paste into another image editor.|