6.3 & 7.5 Image Handling - Making an Extended Focus Image
An extended focus image is formed by taking a z-series stack of images and adding each corresponding pixel of the x-y plane in the z-direction. In effect a sampled volume image is collapsed into a plane. Features in the volume data set are now in focus and in one plane. Extended focus images may be made using the LeicaTCS-NT program ($6.25/15 minutes), which produces poor contrasts or on PC's using tiffsplit4.exe as described below, and in the tiffsplit4.html help pages.
Maximum projection images may also be generated with tiffsplit4.exe or the LeicaTCS-NT software. Maximum projections find the brightest points in the image stack. This process can work amazingly well, especially for fainter structures such a thin dendrites.
Generating extended focus images from sim.tif, a multipaged tiff image files
e.g. Open a DOS box (command prompt) and select the desired drive letter (e.g. e: enter) and path (e.g. cd \users2\fred enter)
Please use tiffsplit4.exe instead of tiffsplit.exe. Tiffsplit4.exe is an enhanced version of tiffsplit.exe.
Extended focus and maximum projection images are automatically contrast normalized and derive their palette from the original TIFF image channels unless -g to convert to grayscale option or -c to remove green/blue under/overload indicator is given. The overlayed image is always in red/green/blue (RGB) mode. Tiffsplit4.exe can also extract the image planes of multipaged tiff files into individual tiff or bitmap format files, which can be opened in general purpose imaging software, such as Adobe Photoshop.
For a summary of tiffsplit4.exe command line switches execute tiffsplit4 without any command line options.
Last updated 10/24/06
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