I need a hardware + software solution for counting and sizing
of dark blue stained colonies on clear plastic plates. The basic
analytical problem and requirements are:
1. Must be easily adaptable for rapid & reproducible counting of many
(50 to 500)wells/dishes of variable size/format : e.g. 35, 60, or 100, or
150 mm dishes; or 6-, 12-, or 24-well plastic tissue culture plates.
2. The number of colonies and the area of each must be obtained in a
form that can be exported to a spreadsheet, or stored in a file.
3. Ability to handle edge refraction lines, by cropping or field
definition. Colony detection near the edges of the plate.
4. Image erosion/dilation, image subtraction, etc are desired, but may
not be essential if the software separates touching colonies .
I have seen several approaches to colony counting, all of which
basically isolate the circular analysis field and perform "spot
counting" and area determination on binary images after thresholding,
etc (e.g. NIH Image). One approach used a video camera to generate
a live image to allow real-time manual positioning of the wells under a
stationary but variable-sized circle superimposed on the monitor,
which defined the analytical field (Artek 880, Image Products Intl.,
Chantilly, VA). This system was a simple video digitizer and gave
only spot count; we also need area, and would like a full featured
image analysis program if possible. Another possible approach could
be by identifying circular areas (best would be to initialize by mouse
point and click to locate areas), followed by edge elimination or find
center and draw radius functions, then analysis of live image or
captured image file. I have also heard of scanner - based methods,
but I believe that this would probably not be practical here. I prefer
the Macintosh platform, but a PC-based system would be acceptable.
Please send replies to this newsgroup, or to my email address :
atown at isnet.is.wfu.edu, or mail to me at : Biochem. Dept., BGSM,
Med. Ctr. Blvd, Winston-Salem, N.C. 27157. Thanks, Alan Townsend.