Distinguishing QD-stained
cellular membranes from PI-stained nucleic
acids in a single cell by tuning the ultrafast delay between IR and
visible pulses. The color bars represent the counts on CCD pixels.
(A) Red-channel PL images of cells costained by both PI dyes and QDs.
(B) MD-WISE image of a costained cell when IR pulse is blocked. The
dashed horizontal line indicates the location where linecut intensity
analysis was performed. (C) Normalized intensity plot aligned along
the line in (B), showing signals throughout the cell (shaded area).
(D) MD-WISE difference PL image with delay t set
to 1 ps, showing no contrast for PI and QD stains. (E) MD-WISE difference
PL image with delay t set to −10 ps, showing
negative contrast for QD-stained membranes. The dashed horizontal
line indicates the location where linecut intensity analysis was performed.
(F) Normalized intensity plot aligned along the line in (E), showing
signal peaks at the location of membranes (shaded area). All the MD-WISE
images in panels B, D, and E were acquired using IR frequency centered
at 2100 cm–1, visible excitation at 550 ± 5
nm, and PL collection wavelength range of 585 ± 18 nm.