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Optical integration
Using the ProgRes® camera instead of the ImagingSource makes no difference concerning camera mounting or the optical overlay procedure. The image on the left shows the setup used for this report, a Nikon Eclipse TE2000 hosting the ProgRes® camera and the JPK NanoWizard®II AFM. The camera is mounted to one of the side ports of the microscope and connected to the JPK NanoWizard®II controller via fire wire. The camera software can then be opened via the camera button within the JPK SPM software. Crucial parameters as exposure time and gain can be adjusted and the fan can be controlled manually, turned off when performing AFM measurements. In case of very sensitive dyes that suffer of fast bleaching there is the possibility to operate a shutter using the input/output trigger option.
To perform the DirectOverlay™ procedure, the cantilever is moved to 25 defined piezo positions and an optical image is taken at each point. The tip locations of these images are then automatically assigned to the corresponding piezo positions. Thus the software can correlate the piezo or rather tip position to the appropriate pixel within the optical image. After this procedure the field of view recorded by the camera is calibrated to the JPK SPM software. At the end of the calibration procedure a snapshot of the sample can be taken or another image taken by the same camera with the same resolution, can be imported. If examining very photosensitive samples it is recommended to take the fluorescence image before the calibration images. Finally interesting regions of the sample can be chosen within the optical image and scanned by AFM.
To perform the DirectOverlay™ procedure, the cantilever is moved to 25 defined piezo positions and an optical image is taken at each point. The tip locations of these images are then automatically assigned to the corresponding piezo positions. Thus the software can correlate the piezo or rather tip position to the appropriate pixel within the optical image. After this procedure the field of view recorded by the camera is calibrated to the JPK SPM software. At the end of the calibration procedure a snapshot of the sample can be taken or another image taken by the same camera with the same resolution, can be imported. If examining very photosensitive samples it is recommended to take the fluorescence image before the calibration images. Finally interesting regions of the sample can be chosen within the optical image and scanned by AFM.