Overview:

Single channel images may be overlayed, in bulk, into red/green channels using bmoverlay.exe.  The images need to be in a format that bmovrlay.exe can use.  Some preparation is required before images can be overlayed as red and green channels.  Images are required to be 8 bit, preferably gray scale bitmaps.  If the image is a time lapse or multiple-channel or z-series tiff image gathered on the LeicaTCS-NT confocal, the images will have been saved as a multipage tiff file.  The individual image pages of the tiff file must be split into separate bitmap format files using tiffsplit4.exe with the -b command line option (see Splitting Multipaged Tiff Files).

The images ideally should be gray scale.  This is best done by using the -g command line option of tiffsplit3.exe.  e.g. tiffsplit4 sim.tif   -g.

Bmovrlay.exe expects images to be supplied in the 8 bit bitmat file format (indexed color or grayscale) with the names supplied to the command line as two matched groups with the same number of files each and with the same pixel dimensions.

The Details:

1. Split the multipage z-series or time-series tiff file into individual bitmap image files using tiffsplit4.exe (dos)
2. Overlay two sets of bitmap images into red/green channels of a bitmap image file (rgb i.e. 24 bit) using bmovrlay.exe (dos)

Last updated 07-31-2003

					© 1998-2003 Michael Chua, Cell & Molecular Physiology, UNC.  All rights reserved.
To Confocal Homepage	To Operating the confocal	To Booking the confocal	Go back a page		Comments: Dr. Michael Chua