Imaging Endogenous Bilirubins with Two-Photon Fluorescence of Bilirubin Dimers.

On the basis of an infrared femtosecond Cr:forsterite laser, we developed a semiquantitative method to analyze the microscopic distribution of bilirubins. Using 1230 nm femtosecond pulses, we selectively excited the two-photon red fluorescence of bilirubin dimers around 660 nm. Autofluorescences from other endogenous fluorophores were greatly suppressed. Using this distinct fluorescence measure, we found that poorly differentiated hepatocellular carcinoma (HCC) tissues on average showed 3.7 times lower concentration of bilirubins than the corresponding nontumor parts. The corresponding fluorescence lifetime measurements indicated that HCC tissues exhibited a longer lifetime (500 ps) than that of nontumor parts (300 ps). Similarly, oral cancer cell lines had longer lifetimes (>330 ps) than those of nontumor ones (250 ps). We anticipate the developed methods of bilirubin molecular imaging to be useful in diagnosing cancers or studying the dynamics of bilirubin metabolisms in live cells.

Efficient near-IR hyperthermia and intense nonlinear optical imaging contrast on the gold nanorod-in-shell nanostructures.

New gold nanorod (Au NR)-in-shell nanostructures were developed to be more efficacious than Au NRs in near-IR (NIR) hyperthermia and nonlinear optical imaging contrast. Au NR-in-shell nanostructures are composed of an intact Au NR in a Au/Ag nanoshell. These nanostructures have a broad, intense absorption band that extends from 400 nm to 900 nm in the NIR. They are more efficient and efficacious than Au NRs with respect to in vitro hypothermia performance. Au NR-in-shell-labeled cancer cells were destroyed using continuous-wave NIR radiation with 50% less laser power than needed for Au NRs. Noticeably, the area of the destroyed cells was significantly larger than the size of the laser irradiation beam; in contrast, the destroyed area was usually restricted to the size of the laser beam spot when Au NRs were used. With their extraordinarily broad and strong surface plasmon resonance band, Au NR-in-shell nanostructures efficiently augmented several multiphoton nonlinear processes as well. The multiphoton emission spectrum covered almost the entire visible wavelength. The yield of the multiphoton signals of Au NR-in-shell nanostructures was on average 55 times larger than that of Au NRs. In vitro images of cancer cells targeted by Au NR-in-shell nanostructures revealed a stronger multiphoton contrast than those targeted by Au NRs.