Hepatycotes in the liver may appear similar in morphology, however, heterogeneities may exist in cellular metabolism.
In this study, in vivo imaging of 6-carboxfluorescein diacetate (6-CFDA) metabolism in the liver was studied. We used
two-photon fluorescence microscopy and hepatic window to provide quantification in studying hepatocellular
metabolism. This model not only provides a potential platform for future study in hepatic responses and regulations, but
also contributes to the fine-tuning of organ-specific functions so as to open up a new era of exciting discoveries.
Acetaminophen (APAP) is the famous drug in global, and taking overdose
Acetaminophen will intake hepatic cell injure. Desptie substantial progress in our
understanding of the mechanism of hepatocellular injury during the last 40 years,
many aspects of the pathophysiology are still unknown or controversial.1 In this study,
mice are injected APAP overdose to damage hepatocyte. APAP deplete glutathione
and ATP of cell, N-Acetyl Cysteine (NAC) plays an important role to protect
hepatocytes be injury. N-Acetyl Cysteine provides mitochondrial to produce
glutathione to release drug effect hepatocyte. By 6-carboxyfluorescein diacetate (6-CFDA) metabolism in vivo, glutathione keep depleting to observe the hepatocyte
morphology in time. Without NAC, cell necrosis increase to plasma membrane
damage to release 6-CFDA, that's rupture. After 6-CFDA injection, fluorescence will
be retained in hepatocyte. For cell retain with NAC and without NAC are almost the
same. With NAC, the number of cell rupture decreases about 75%.
Combining multiphoton microscopy with a newly designed hepatic imaging window, we acquired
in vivo images of mice obstructive cholestasis. We observed that in mice with bile duct ligation, bile
canaliculi failed to appear during the whole observation period over 100 minutes following
carboxyfluorescein diacetate injection, whereas the fluorescence was retained much longer within
sinusoids. Furthermore, the fluorescence intensities in sinusoids were persistently higher than in
hepatocytes during the course.
In this work, we utilized multiphoton microscopy for the label-free diagnosis of non-cancerous, lung adenocarcinoma
(LAC), and lung squamous cell carcinoma (SCC) tissues from human. Our results show that the combination of second
harmonic generation (SHG) and multiphoton excited autofluorescence (MAF) signals may be used to acquire
morphological and quantitative information in discriminating cancerous from non-cancerous lung tissues. Specifically,
non-cancerous lung tissues are largely fibrotic in structure while cancerous specimens are composed primarily of tumor
Quantitative ratiometric analysis using MAF to SHG index (MAFSI or SAAID) shows that the average MAFSI for noncancerous
and LAC lung tissue pairs are 0.55 ±0.23 and 0.87±0.15 respectively. In comparison, the MAFSIs for the noncancerous
and SCC tissue pairs are 0.50±0.12 and 0.72±0.13 respectively. Intrinsic fluorescence ratio (FAD/NADH) of
SCC and non-cancerous tissues are 0.40±0.05 and 0.53±0.05 respectively, the redox ratio of SCC diminishes
significantly, indicating that increased cellular metabolic activity.
Our study shows that nonlinear optical microscopy can assist in differentiating and diagnosing pulmonary cancer from
non-cancerous tissues. With additional development, multiphoton microscopy may be used for the clinical diagnosis of
We utilize multiphoton microscopy for the label-free diagnosis of noncancerous, lung adenocarcinoma (LAC), and lung squamous cell carcinoma (SCC) tissues from humans. Our results show that the combination of second-harmonic generation (SHG) and multiphoton excited autofluorescence (MAF) signals may be used to acquire morphological and quantitative information in discriminating cancerous from noncancerous lung tissues. Specifically, noncancerous lung tissues are largely fibrotic in structure, while cancerous specimens are composed primarily of tumor masses. Quantitative ratiometric analysis using MAF to SHG index (MAFSI) shows that the average MAFSI for noncancerous and LAC lung tissue pairs are 0.55±0.23 and 0.87±0.15, respectively. In comparison, the MAFSIs for the noncancerous and SCC tissue pairs are 0.50±0.12 and 0.72±0.13, respectively. Our study shows that nonlinear optical microscopy can assist in differentiating and diagnosing pulmonary cancer from noncancerous tissues.
Conventional hepatic research relies heavily on histological images for obtaining morphological
information of the liver. However, static histological images can not provide real-time dynamic
information of in vivo physiological processes such as cellular motion or damage. For a long time,
panadol has been used in pain relief. However, Panadol may have unwanted side effects and detailed
information of the effects of Panadol on hepatic metabolism is unknown. In this work, we developed
a high resolution intravital hepatic imaging chamber to study the effects of Panadol on liver. We
expect this methodology to be useful in revealing the detailed metabolism of liver after using Panadol
and this approach allows us to achieve a better understanding of hepatic processes. In our approach,
we use multiphoton fluorescence (MPF) microscopy to observe the side effect of liver on using
Panadol inside the in vivo mouse animal model.
Bile is the exocrine secretion of liver and synthesized by hepatocytes. It is drained into duodenum for the function of
digestion or drained into gallbladder for of storage. Bile duct obstruction is a blockage in the tubes that carry bile to the
gallbladder and small intestine. However, Bile duct ligation results in the changes of bile acids in serum, liver, urine, and
feces1, 2. In this work, we demonstrate a novel technique to image this pathological condition by using a newly
developed in vivo imaging system, which includes multiphoton microscopy and intravital hepatic imaging chamber. The
images we acquired demonstrate the uptake, processing of 6-CFDA in hepatocytes and excretion of CF in the bile
canaliculi. In addition to imaging, we can also measure kinetics of the green fluorescence intensity.
In this investigation, we used <i>in vivo</i> nonlinear optical microscopy to image normal and carcinogen DMBA treated skin
tissues of nude mice. We acquired two-photon autofluroescence and second harmonic generation (SHG) images of the
skin tissue, and applied the ASI (Autofluorescence versus SHG Index) to the resulting image. This allows us to visualize
and quantify the interaction between mouse skin cells and the surrounding connective tissue.
We found that as the imaging depth increases, ASI has a different distribution in the normal and the treated skin tissues.
Since the DMBA treated skin eventually became squamous cell carcinoma (SCC), our results show that the
physiological changes to mouse skin en route to become cancer can be effectively tracked by multiphoton microscopy.
We envision this approach to be effective in studying tumor biology and tumor treatment procedures.
Liver is the chemical factory in body responsible for important functions such as metabolism and
detoxification. When liver can not be regenerated in time to amend damages that has occurred, failure
of hepatic functions can result. Traditionally, the study of liver pathology has depended on histological
techniques, but such methods are limited to ex-vivo observation. In order to study hepatic metabolism
in vivo, we have designed a hepatic imaging chamber made of biocompatible titanium alloy (6V4Al-Ti, ELI grade). In combination with multiphoton and second harmonic generation microscopy, our
approach allows the intravital observation of hepatic intravital activities to be achieved. Processes such as hepatic metabolism and disease progression can be studied using this methodology.