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Fig.
2. Simultaneous detection of many genes. (A) A single human
colon adenocarcinoma cell (DLD-1) G2 nucleus (by DAPI signal)
with a pseudo-colored representation of 17 transcription sites
detected in situ. The image is "flattened" such that all 12 0.5-µm
Z-sections are displayed on the background, which is the DAPI counterstain
from the middle image of the stack. Gene identity is denoted by color
and the Z-location is recorded by the adjoining number. Lower numbers
represent closer proximity to the cover slip. Scale bar, 3 µm.
(B) Three G1 DLD-1 nuclei from the same field, which
together express all 10 genes assayed. Arrows indicate sites that
are shown below magnified from the original data. From left to right,
the 10 marked transcription sites are IL-8, MCL-1, DUSP-1, cyclin
D1, g-actin, EGR-1, TIEG-1, ß-actin,
c-myc, and c-jun. Scale bar, 3 µm. (C) Chart of the
10 genes detected in (B). The "Pseudo" column shows the arbitrary
pseudo-color used to denote the gene identity of each transcription
site in the renderings above. Columns at right show the actual signal
recorded at the appropriate Z-section for the transcription sites shown
with an arrow in (B). Each band of data of fluorescein isothiocyanate
(FITC), Cy3, Cy3.5, and Cy5 is shown, with the positively scored signals
highlighted by surrounding boxes. Each area of the unprocessed image
shown is 1 µm2. Observed misalignment is due to
chromatic shift between filter sets. |
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