Diagnosing hepatocellular carcinoma with the intensity and the lifetime of two-photon red autofluorescences

Liu, Tzu-Ming; Hsieh, Chien-Tai; Chen, Yu-Shing; Huang, Fu-Lien; Huang, Hsin–Yi; Lee, Wei-Jei; Kung, Chun-Ta; Sun, Chi‐Kuang

doi:10.1117/12.872690

Cited by 4 publications

(8 citation statements)

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“…However, their 2.3 ns fluorescence lifetime (Supporting Information Figure S2C) is much longer than the 100−400 ps 2PAF lifetimes observed in fresh liver tissues. 46 These results indicate that biliverdin and porphyrin have relatively rather small contribution on this red autofluorescence. Therefore, we thought this characteristic red fluorescence might be related to bilirubins and/or their derivatives and have potential to be a useful contrast for bilirubin molecular imaging.…”

mentioning

confidence: 92%

“…The possible candidates were limited to porphyrins, biliverdins, or bilirubins. 46 Through the fluorescence spectrum and lifetime measurement on standard chemicals, obviously, porphyrins (Supporting Information Figure S1) are quite different from those measured in liver tissues ( Figure 1). The emission peaks of HSA-associated biliverdins are both at 690−700 nm under single-photon and multiphoton excitation (Supporting Information Figure S2, parts A and B), which is close to the 660 nm emission peak ( Figure 1).…”

mentioning

confidence: 98%

“…Recently, we found liver, a bilirubin-rich organ, has a 660 nm two-photon autofluorescence (2PAF) excited at 1230 nm (Figure ). Far away from two-photon excitation of Soret bands, most of endogenous fluorophores will not be excited under this excitation condition. The possible candidates were limited to porphyrins, biliverdins, or bilirubins .…”

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confidence: 99%

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Imaging Endogenous Bilirubins with Two-Photon Fluorescence of Bilirubin Dimers

Shen

Tsai²,

Chen³

et al. 2015

Anal. Chem.

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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.

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confidence: 92%

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confidence: 98%

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Imaging Endogenous Bilirubins with Two-Photon Fluorescence of Bilirubin Dimers

Shen

Tsai²,

Chen³

et al. 2015

Anal. Chem.

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“…55 HCC can be identified by the reduced intensity levels and longer fluorescence lifetime. 56 Similarly, it has been suggested that the autofluorescence can be used to assist discrimination of HCC grade 1 due to significantly higher intensity in nontumor (NT) tissue. Therefore, one group sought to develop multiphoton autofluorescence (MAF) as an effective label-free tool in diagnosing HCC in ex vivo tissue.…”

Section: Cancermentioning

confidence: 99%

“…58 The usability of TPEF in diagnosing HCC has also been done in combination with FLIM. 52,56 FLIM was used to assess molecular events in vivo in the whole-body using a tumor-specific NIR molecular probe. 59 Most of these studies diagnosing liver fibrosis and cancer have been done by utilizing the cell's autofluorescent properties.…”

Section: Cancermentioning

confidence: 99%

Multiphoton microscopy in defining liver function

et al. 2014

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Multiphoton microscopy is the preferred method when in vivo deep-tissue imaging is required. This review presents the application of multiphoton microscopy in defining liver function. In particular, multiphoton microscopy is useful in imaging intracellular events, such as mitochondrial depolarization and cellular metabolism in terms of NAD(P)H changes with fluorescence lifetime imaging microscopy. The morphology of hepatocytes can be visualized without exogenously administered fluorescent dyes by utilizing their autofluorescence and second harmonic generation signal of collagen, which is useful in diagnosing liver disease. More specific imaging, such as studying drug transport in normal and diseased livers are achievable, but require exogenously administered fluorescent dyes. If these techniques can be translated into clinical use to assess liver function, it would greatly improve early diagnosis of organ viability, fibrosis, and cancer.

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Visualizing liver anatomy, physiology and pharmacology using multiphoton microscopy

Wang

Liang

Gravot

et al. 2016

Journal of Biophotonics

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Multiphoton microscopy (MPM) has become increasingly popular and widely used in both basic and clinical liver studies over the past few years. This technology provides insights into deep live tissues with less photobleaching and phototoxicity, which helps us to better understand the cellular morphology, microenvironment, immune responses and spatiotemporal dynamics of drugs and therapeutic cells in the healthy and diseased liver. This review summarizes the principles, opportunities, applications and limitations of MPM in hepatology. A key emphasis is on the use of fluorescence lifetime imaging (FLIM) to add additional quantification and specificity to the detection of endogenous fluorescent species in the liver as well as exogenous molecules and nanoparticles that are applied to the liver in vivo. We anticipate that in the near future MPM‐FLIM will advance our understanding of the cellular and molecular mechanisms of liver diseases, and will be evaluated from bench to bedside, leading to real‐time histology of human liver diseases.

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Diagnosing hepatocellular carcinoma with the intensity and the lifetime of two-photon red autofluorescences

Cited by 4 publications

References 0 publications

Imaging Endogenous Bilirubins with Two-Photon Fluorescence of Bilirubin Dimers

Imaging Endogenous Bilirubins with Two-Photon Fluorescence of Bilirubin Dimers

Multiphoton microscopy in defining liver function

Visualizing liver anatomy, physiology and pharmacology using multiphoton microscopy

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