Multi-point Focus

requires amics 5 1 0 or newer the released version of amics is and can be found at amics 5 1 release notes docid\ se8jyhxszdqfpit7s1ja0 with instruction on the software installation docid\ e1faehd1k20uedqhptzlx page manuals for previous versions of the software can be found at the older amics versions docid\ d2biwe sybowhhdww0anz page a good sample focus is critical for accurate segmentation results in some cases, for example, if the sample area to analyze is quite large, it is possible that the focus position is not the same at all points across the sample for such cases, a 2 point (for long, narrow areas) or 3 point focus procedure can be used to ensure the entire sample area is in focus when measured note! the focus difference across the sample should not be more than 2mm step 1 in investigator right click and record the hardware conditions with get hw data step 2 right click again and select manage hw data… step 3 the hw data manager dialog will open as shown to the right the red cross indicates the first recorded focus point for large areas, a further two focus positions are required, which for best results should be spaced as far apart as possible and define the corners of a triangular shape step 4 right click in the display on the position to use as a second focus point and select hw to position step 5 using the microscope controls, set the focus at this position (in this case only focus is recorded by amics so the magnification does not need to be set back to the measurement conditions setting) switch back to amics investigator and click on it step 6 repeat steps 4 and 5 to record a third position note that the three positions will be indicated in the display and can be deleted or updated if necessary step 7 once three positions have been recorded, evaluate the focus select any point on the sample, right click, and select hw to position step 8 once the sem stage has moved to the position, right click again and select set wd hereto set the sem stage to the calculated working distance (wd) the sem image should be set to a focussed image if satisfied with the result, the hw data manager dialog can be closed step 9 in the amics investigator display the three focus positions will still be indicated the sample can be measured as usual below is an example video showing how to set up a multipoint focus in amics