LEICA ACQUIRE WINDOWS WINDOWS
Note that the bars cannot be moved past each other (a reflection of the physical arrangement of the detectors and sliders), so if you are simultaneously scanning with one than one laser and using multiple detectors, you need to use the windows on the left/center for the shorter wavelengths, and the center/right windows for collecting longer wavelength light. With 5 bars things can get confusing, so look for the grey lines that link the windows to the control bars for each detector to keep track of which one your are adjusting. The laser lines and emission curves are good guides for setting the spans of the emission windows. You can right click again to remove either curve if you wish. Clicking on “Show Excitation Graph” adds a dotted line excitation curve: To see the excitation curve, right click on the spectra display to get this new window. When a dye is selected, an emission curve (solid line) will appear: You can do a lambda scan of any unlisted dye and add it to the drop down menu. If your dye is not listed, choose the closest match. Click on the top drop down menu of a detection window and select your fluor: They are not required to scan, but are very helpful for optimal setting of the slider bars.
LEICA ACQUIRE WINDOWS SOFTWARE
The software also allows a user to display the emission and/or excitation curves for the fluors detected. When a laser is active and ready to use, a corresponding line will appear on the spectrum diagram: When the detector is active, this window diagram will be colored to reflect the portion of the spectrum it is set to detect. The yellow arrow indicates the detection window settings, which control the wavelengths of light allowed into the detector. 3 controls Gain (power to the detector), 4 controls Offset, and 5 is a drop down menu that allows you to display emission-excitation spectra on the visible spectrum diagram. The controls for each on these detectors are arranged within the horizontal grey bars.ġ is the button that allows you to select the channel color. The lower region diagrams the visible spectrum at the top controls for the 5 fluorescence detectors and a transmitted light PMT are underneath. There are two versions of the laser panel, and you can switch between them using the button on the right (*): You can click and drag, use the scroll wheel on the mouse, or click on the number above a bar and type in the desired value. If a laser has not been turned on via the laser configuration panel, the bars will be greyed and cannot be adjusted. There are on switches for both the UV and visible light lasers, and slider bars for adjusting the power of each laser line. The top portion is used to control the laser intensity. This is the central region of the Leica acquisition panel. This is why if you are using only 1 fluorescent detector, and you do not required the extra-sensitivity of the HyD, you should use PMT3, because it can directly “see” all the wavelengths. Those sliders will admit selected wavelengths and reflect the remainder to the last two detectors (PMT1 & 5). They allow light of the specified wavelength range into the detector, and reflect the blocked wavelengths to the next two detectors (HyD2 & 4). The position of the sliders (2) is adjusted from the acquisition panel. The collected light is sent to PMT3 first.
This is a diagram of their arrangement in the hardware of the system: Our SP8 has 3 photo-multiplier tubes (PMTs) and 2 Hybrid (HyD) detectors. Most of the central portion of the acquisition panel is devoted to the control of the lasers that help generate the photons and the detectors that collect them. Setting up your detection windows in the SP8 software
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