Smart Setup Dialog

1	+ Dye & Contrast Methods Opens the Add Dye or Contrasting Method dialog, see Configuring Multi-Channel Acquisition with Smart Setup.
2	Table with selected channels You can add up to 8 fluorescence channels and contrast methods. The added dyes or the contrast methods are shown in the list. If necessary, change the color by clicking the arrow and select a user defined color.
3	Buttons to optimize Acquisition Parameters Click on a button to optimize image acquisition regarding particular requirements and to influence parameters like the camera, detector, and lighting settings, see Buttons to Optimize Acquisition Parameters.
4	Proposals for Specific Filter Settings Displays various Smart Setup proposals for further experiment settings. See Graphical Display of Proposals and Smart Setup Proposals.
5	Show Excitation checkbox Activated: Shows the excitation spectrum of the selected dyes in the graphical display.
6	Show Emission checkbox Activated: Shows the emission spectrum of the selected dyes in the graphical display.

Buttons to Optimize Acquisition Parameters

The automatic settings you make through these buttons affect the settings displayed in the Channels tool, which are used for acquisition, see Channels Tool.

Parameter	Description
Automatic	Sets power of Colibri-LEDs depending on wavelength and power of transmitted and reflected light sources to 10% or 3V. The system tries to set the optimal resolution for the camera by adjusting the binning if supported. The resolution is calculated from the total magnification at the camera adapter (objective + optovar + adapter). Sets the Exposure Time: Transmitted light channel: 10 ms Reflected light channel: 10 ms Fluorescence channel: 150 ms Sets the Auto Exposure to Off Sets Intensity to the following: Transmitted light channel: 70% Reflected light channel: 70% Fluorescence channel: 30%
Speed	Sets power of Colibri-LEDs 100% and power of transmitted and reflected light sources to 10% or 3V. The system tries to set the optimal resolution for the camera by adjusting the binning if supported. The resolution is calculated from the total magnification at the camera adapter (objective + optovar + adapter). Sets the Exposure Time: Transmitted light channel: 3 ms Reflected light channel: 3 ms Fluorescence channel: 50 ms Sets the Auto Exposure to Off Sets the Intensity to the following: Transmitted light channel: 30% Reflected light channel: 30% Fluorescence channel: 15%
Signal	Sets power of Colibri-LEDs 75% and power of transmitted and reflected light sources to 10% or 3V. The system tries to set the optimal resolution for the camera by adjusting the binning if supported. The resolution is calculated from the total magnification at the camera adapter (objective + optovar + adapter). Sets the Exposure Time: Transmitted light channel: 20 ms Reflected light channel: 20 ms Fluorescence channel: 300 ms Sets the Auto Exposure to Off Sets the Intensity to the following: Transmitted light channel: 100% Reflected light channel: 100% Fluorescence channel: 60%
Default	Sets power of Colibri-LEDs 10% and power of transmitted and reflected light sources to 10% or 3V. The system tries to set the optimal resolution for the camera by adjusting the binning if supported. The resolution is calculated from the total magnification at the camera adapter (objective + optovar + adapter). Sets the Exposure Time to 20ms. Sets the Auto Exposure to Default Sets the Intensity to 100%.
Current	NOTICE! If you changed hardware settings manually and do not want to lose them, click Current. Sets power of Colibri-LEDs depending on wavelength and power of transmitted and reflected light sources to 10% or 3V. The system tries to set the optimal resolution for the camera by adjusting the binning if supported. The resolution is calculated from the total magnification at the camera adapter (objective + optovar + adapter). Transfers the following values from the current camera in use to the Channels tool: Auto Exposure Exposure Time Intensity

Smart Setup Proposals

The number and type of proposals depend on the used microscope hardware, the selected dyes, and the contrast method.

Parameter	Description
Best Signal	Results in the best signal strength.
Fastest	Results in the fastest acquisition.
Best Compromise	Results in the best compromise between signal strength and acquisition speed.

Graphical Display of Proposals

The bars in the graphs only show relative values. The actual strength of the emission signal and the crosstalk in the image can deviate substantially from this estimate, as Smart Setup has no knowledge of the strength with which the sample has been dyed with the individual dye components.

Emission Signal, Speed, Crosstalk, and Tracks

1	Emission Signal A filled, colored bar in the Emission Signal display field shows the relative emission signal to be expected for the corresponding channel. The channel color corresponds to the color of the selected dye in the Configure Experiment section.
2	Speed A gray bar in the Speed display field represents the approximate acquisition speed that can be expected. This is the time required for the movement of microscope hardware during multichannel acquisition. Camera exposure times or parameters for other acquisition dimensions are not taken into account here.
3	Crosstalk A hatched bar in the Crosstalk display field shows the expected relative crosstalk originating from one or more dyes for other channels.
4	Tracks display Only visible if the Show Excitation and/or Show Emission checkboxes are activated. The various tracks are labeled with T1, T2 etc. The white lines show the excitation and emission spectra of the dyes schematically. The spectra are filled in color in the places that will be acquired by the acquisition configuration suggested by Smart Setup. Transmitted light channels are displayed as a white field.

Smart Setup Dialog

Buttons to Optimize Acquisition Parameters

Smart Setup Proposals

Graphical Display of Proposals

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