ISSN:
1089-7623
Source:
AIP Digital Archive
Topics:
Physics
,
Electrical Engineering, Measurement and Control Technology
Notes:
A microprocessor-controlled system has been developed to automatically scan tissue culture dishes in order to detect and locate fluorescently labeled cell colonies for subsequent cloning. An X-Y recorder mechanism moves an 80-mm-diam dish in a raster scan through a stationary laser beam adjacent to a photomultiplier tube equipped with an emission filter. Front-end electronics processes the PMT signal to screen out small-scale fluorescent artifacts of selectable size (e.g., 〈0.2 mm). Appropriate signals are input to the computer which then interrupts the raster scan to perform a limited fine scan of the dish to accurately localize the fluorescent spot position. An additional artifact test using a different filter is then performed. The underside of the dish is scribed at the location of spots that pass this test. Using fluorescein as a label, fluorescence as low as 25% above intrinsic cell background fluorescence can be detected. The fluorescence signal level threshhold can be set (e.g., 70% above intrinsic cell background) to within an accuracy of approximately 15% of intrinsic cell background fluorescence, and the system will reliably find colonies with a signal exceeding that level. With the above typical values, the number of false positives is typically less than five per dish and the time to completely scan an 80-mm-diam tissue culture dish is 2–4 min.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1063/1.1140303
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