Vh. Latham et al., DESK-TOP COMPUTER-BASED IMAGE-ANALYSIS OF CELL-SURFACE FLUORESCENCE PATTERNING FROM A PHOTOGRAPHIC SOURCE, Acta histochemica, 98(3), 1996, pp. 295-300
We report the use of standard computer-based image analysis technology
to analyze, from a photographic source, individual cell surface recep
tor site patterns using fluorochrome labeling. The model used in this
study was a Strongylocentrotus purpuratus sea urchin embryo labeled wi
th fluorescein isothiocyanate-conjugated wheat germ agglutinin (FITC-W
GA) (0.5 mg/ml for 5 min at 15 degrees C incubated with fertilization
membrane free embryos). Image capture was performed using desktop-type
digital scanning, and the images were imported into Adobe Photoshop f
or study. All images analyzed were derived from a single photographic
negative: 1) the actual micrograph printed from the negative and scann
ed into a Macintosh IIx computer; 2) the scanned negative itself; and,
3) a high resolution scanning process used with a Kodak Photo CD. Pat
terns were analyzed using the densitometry feature of Photoshop, and w
ere similar enough from all 3 scanned images to indicate that any of t
he 3 scanning processes can be used for fluorescence patterning analys
is. Adobe Photoshop was also used to highlight, for closer analysis, t
he fluorescence patterns by producing 3-D effects, border mapping and
transition area detailing. The desktop image analysis procedures descr
ibed here to study fluorescence patterning require no expensive scient
ific hardware or software.