CHAPTER 1 - SYSTEM OPERATIONZEISS Left Tool Area and Hardware Control Tools LSM 880148 000000-2071-464 10/2014 V_015.2.15.5 CorrectionExpand the Correction field by pressing thetriangle (Fig. 198). The Refractive IndexCorrection input box allows you to consider thedifferent refractive indices between the immersionmedium of the objective (n') and the embeddingmedium of the specimen (n), which can be setbetween 0.5 and 3 by typing or using the arrowkeys. The number should correspond to the ratior=n/n’.Auto Z Brightness Correction (Fig. 199) allowssetting of different values for the scan parametersGain (Master), Digital Offset, Digital Gain andLaser Attenuation of up to 10 freely selectableslices within a Z-Stack. During the scan procedure,the values of these parameters are automaticallylinearly interpolated between the neighboringpositions within the Z-Stack unless Splineinterpolation is selected. This function can beactivated and deactivated with the check box UseCorrection.Add takes the above mentioned parameters andcorrelates them with the current Focus Positionwhich is then added as a Position in the list (forexample the first slice of a stack).Adding a new position with a defined set of channel parameters cannot be done while scanning. Thepositions don't need to be listed in a specific order, the program will sort the values automatically in thebackground.Move to moves the actual focus to the highlighted Position in the list without scanning. Starting aContinuous scan then allows setting or correcting the parameters and storing them again hitting Add.Highlighted positions can be removed (Remove) or the whole list can be deleted (Remove all).The interpolation between the Positions in the list can be extrapolated (check the box next toExtrapolate) to the actual first and last slice of a Z-Stack if those are not part of the range of thePositions in the list.When Enable Test is active, the settings of the Position list or the interpolated values calculated for therespective focus position are applied while scanning at single positions of the stack or while focusingthrough the stack using the Focus tool or the focus drive of the microscope. This allows controlling thesettings before starting the actual acquisition of the stack.A position list can be saved and loaded again for further experiments. Use the Save… and Load…button accordingly. Make sure the Z reference position of the list matches the reference position of theactual Z stack. It is highly recommended to set the Z focus to zero (ideally at the first or last slice of thestack) when defining the stack and the correction positions within the Z stack. For a subsequentexperiment the position zero than has to be defined at the position of the first or last slice respectively.Then the correction parameters will be applied in the desired way. Setting the focus position to zero doesnot change the position values of the list!Fig. 198 Correction field – appearance forthe WF, SIM and Laser WF imagingmodesFig. 199 Auto Z Brightness Correction
