CHAPTER 1 - SYSTEM OPERATIONZEISS Left Tool Area and Hardware Control Tools LSM 880352 000000-2071-464 10/2014 V_015.4.17.3 Structured Illumination – TIRF-FilteredThis function works for SIM images (stack) incombination with a TIRF image. The lowest frame(first frame) of the stack, preferentially one takenclose to the coverslip, will be taken and combinedwith the TIRF. The SIM image contributes the x,yenhancement, the TIRF image the z-resolution ofthe TIRF.Select the TIRF-Filtered tab to select the TIRF-Filtered Processing tool (Fig. 497). Activate theSIM image you want to process in the imagecontainer and press the Select button for Inputimage to load the SIM image. Activate the TIRFimage you want to use as the reference in theimage container and press the Select button forInput image 2. Then expand the MethodsParameters tool window to have access to settingparameters (Fig. 498).You can toggle between Input image 1 and 2 inthe Preview tab window, which also displays apreview of the output image.The Image input description field lists all thechannels in the SIM column, the used lasers of achannel in the Laser column and the employedcamera coded by a number in the Cameracolumn.The Image input 2 description field lists all thechannels in the TIRF column and the used laser ofa channel in the Laser column.It is important to load the SIM image asInput image and the TIRF image as Inputimage 2 as the software expects it that way.If in Input image 2 an image stack isselected, the system gives an error messageas it expects a single plane (Fig. 499).Fig. 497 Structured illumination – TIRF-Filtered tool selectedFig. 498 Structured Illumination – TIRF-Filtered toolFig. 499 Error message for falsely loadedimage dimensions in Input image 2
