Zeiss LSM 710 SIM Manual/quick Manual
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Manual processingIf SIM artifacts and aberrations are visible in the processed image, fine tuning with manualprocessing can help to eliminate the problems.During the SIM introduction we discuss the parameters and how to find/remove artifacts in moredetails.For the best results acquire always a small Z-stack (use Center mode) with around 5 planes withoptimal section intervals. With this small focus drifts can be also avoided.Don’t forget to save the file after acquisition and after processing. Use .czi format and if you need,export the image as .tiff.Fourier space (dummy, notfrom the exact dataset, butgood for checking the effectof the different settings)First check what parameterswere used for automaticprocessing under the info tab.Noise filter has the biggesteffect in changing theresolution and removingartifactsBaseline shifted/cut:Background values for thehistogram. “Shift” keepseverything, “cut” contrasts,removes backgroundSwitch to manual processingto change parametersSelect the planes to process(single plane or first-last)If it is a Z-stack, select 3DUncheck Auto noise filterand change the Noise filterparameter. If the automaticprocessing was -6, try todecrease the resolution to -5,-4, etc till SIM artifactseliminated. Don’t change theSR Frequency Weighting.Channel selection forprocessing |
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