Superior pathological diagnosis using transparent tissues

September 13, 2017
Fig.1. Tissue clearing of various human tissues with CUBIC. (Upper) Gross and microscopic images of human lung and lymph nodes cleared by CUBIC. After gross image acquisition at the indicated time points, lung and lymph node tissues were washed with PBS, followed by paraffin embedding, sectioning, and H&E staining. These images show that lung and lymph node tissues were significantly cleared and that CUBIC clearing caused only negligible, if any, degeneration of the tissues. (Lower) Tissue blocks from formaldehyde-fixed normal human lung and lymph node were cut in half. One half was kept in PBS (“Not embedded”) whereas another half was first embedded into a paraffin block, was recovered by deparaffinization (“Paraffin-embedded”), and then was subjected to tissue clearing by CUBIC. The gross appearance of the resulting samples was not distinguishable and had an equivalent transparency. Credit: Nojima S. et al. Scientific Reports

RIKEN Quantitative Biology Center and Osaka University researchers show CUBIC, a tissue clearing and 3-D imaging technique, makes human organs transparent to improve pathology diagnosis

To some, the idea of invisibility leads to mischief, seeing things without being seen. To the pathologist, the idea of invisibility leads to simplicity, seeing the disease in its fullest. In a new study published in Scientific Reports, Japanese researchers report, CUBIC, a technique including a tissue processing that makes transparent, provides better assessment of lesions for pathological diagnosis.

"Traditionally, pathological diagnosis is made by taking 2-D sections of a specimen resected from patients. It is effective, but we cannot exclude the possibility that important findings away from the cut surface are overlooked," explains Osaka University Professor Eiichi Morii, who co-corresponding-authored the study.

Contemporary methods are based on staining techniques from the 19th Century. Patient specimens are cut into thin sections that are stained and analyzed individually under a microscope. However, this method has limitations in its relatively narrow range of observation area and in its two-dimensionality. CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational Analysis) was first reported by RIKEN Group Director Hiroki R. Ueda, who is the corresponding author of this study, and his colleagues three years ago. CUBIC has been used to observe whole organs mainly from experimental animals. In the new study, Ueda and his colleagues demonstrated that CUBIC can be used to observe organs from humans and that it surpasses current methods for pathological diagnosis/study.

Fig.2. 3D imaging of pathological specimens with CUBIC. (Left) The reconstructed 3D image of a normal human lung tissue stained with green-fluorescent nucleic acid stain SYTO 16 and Alexa Fluor 647-conjugated anti-α-SMA (smooth muscle actin) antibody, showing the network of vasculature in the pulmonary alveolar interstitium. The image was obtained by confocal fluorescence microscopy. (Middle) The reconstructed 3D image of a lung tissue derived from an amyloidosis patient. The tissue was stained with SYTO 16 and Congo Red, showing the amyloid deposition in artery wall in lung. The image was obtained by confocal fluorescence microscopy. (Right) The reconstructed 3D image of SYTO 16-stained half-cut human mesenteric lymph node. The image was obtained by light-sheet fluorescence microscopy. Credit: Nojima S. et al. Scientific Reports

The study shows CUBIC applicability to the 3-D imaging of patient lung and lymph node tissues, clearly delineating normal and abnormal regions (Figure 1, Figure 2). After routine observation, many patient samples are stored at hospitals as paraffin-embedded blocks. Additionally, the study shows that the combination of appropriate deparaffinization and CUBIC enabled 3-D imaging of these older specimens.

"These results mean that we can use not only newly fixed samples but also paraffin-embedded tissues stored in the pathological archives of hospitals," said Osaka University Assistant Professor Satoshi Nojima, who first-authored the study.

The scientists also examined the practical diagnostic potential of CUBIC. They showed that CUBIC was much more capable of detecting metastatic carcinomas in lymph node specimens compared to standard pathology techniques (Figure 3).

Fig.3. Screening of metastatic carcinoma nodules in lymph nodes with CUBIC. (Upper) Schematic diagram of the metastasis screening protocol. (Middle) The representative images of reconstructed 3D model, H&E staining, or Cytokeratin immunohistochemical (IHC) staining of the lymph node contained small carcinoma nodules which were only identified by CUBIC screening but were not detected by routine diagnosis method. (Lower) The results of screening are summarized in the table. Four lymph nodes out of 74 were newly diagnosed as positive for metastasis only by the CUBIC-based screening, making the sensitivity of the conventional diagnosis method 85.2% (23/27) with four false negative, i.e. 14.8% (4/27) improvement in sensitivity with CUBIC. Credit: Nojima S. et al. Scientific Reports

"This is outstanding result to demonstrate the usefulness of CUBIC on practical clinical examination," Nojima said.

These findings show the potential of CUBIC for retrospective and prospective clinicopathological .

"Our wish is to improve CUBIC so that it leads to the establishment of a novel field of medical science based on 3-D histopathology." Ueda said.

Explore further: Watch cancer spread in a mouse

More information: Satoshi Nojima et al. CUBIC pathology: three-dimensional imaging for pathological diagnosis, Scientific Reports (2017). DOI: 10.1038/s41598-017-09117-0

Related Stories

Watch cancer spread in a mouse

July 5, 2017
Researchers in Japan have developed a method to image cancer at the single-cell level by using chemical techniques to make whole mouse bodies and organs highly transparent. Combining their preparation with existing imaging ...

Japan scientists make see-through mice

November 6, 2014
Researchers at the RIKEN Quantitative Biology Center in Japan, together with collaborators from the University of Tokyo, have developed a method that combines tissue decolorization and light-sheet fluorescent microscopy to ...

Rapid whole-brain imaging with single cell resolution

April 17, 2014
A major challenge of systems biology is understanding how phenomena at the cellular scale correlate with activity at the organism level. A concerted effort has been made especially in the brain, as scientists are aiming to ...

'Expansion pathology' method could mean earlier intervention

July 21, 2017
(HealthDay)—A new method, called expansion pathology (ExPath), which is a clinically optimized form of expansion microscopy (ExM), can be used for pathology and clinical research, according to a report published online ...

Improved imaging of neonatal soft-tissue tumors can help radiologists improve patient care

July 25, 2017
Better understanding of practical imaging techniques with regard to neonatal soft-tissue tumors can improve patient care, according to an article published in the July 2017 issue of the American Journal of Roentgenology (AJR).

Recommended for you

Hibernating ground squirrels provide clues to new stroke treatments

November 17, 2017
In the fight against brain damage caused by stroke, researchers have turned to an unlikely source of inspiration: hibernating ground squirrels.

Molecular guardian defends cells, organs against excess cholesterol

November 16, 2017
A team of researchers at the Harvard T. H. Chan School of Public Health has illuminated a critical player in cholesterol metabolism that acts as a molecular guardian in cells to help maintain cholesterol levels within a safe, ...

Prototype ear plug sensor could improve monitoring of vital signs

November 16, 2017
Scientists have developed a sensor that fits in the ear, with the aim of monitoring the heart, brain and lungs functions for health and fitness.

Ancient enzyme could boost power of liquid biopsies to detect and profile cancers

November 16, 2017
Scientists are developing a set of medical tests called liquid biopsies that can rapidly detect the presence of cancers, infectious diseases and other conditions from only a small blood sample. Researchers at The University ...

FDA to crack down on risky stem cell offerings

November 16, 2017
U.S. health authorities announced plans Thursday to crack down on doctors pushing stem cell procedures that pose the gravest risks to patients amid an effort to police a burgeoning medical field that previously has received ...

Engineering the gut microbiome with 'good' bacteria may help treat Crohn's disease

November 15, 2017
Penn Medicine researchers have singled out a bacterial enzyme behind an imbalance in the gut microbiome linked to Crohn's disease. The new study, published online this week in Science Translational Medicine, suggests that ...

0 comments

Please sign in to add a comment. Registration is free, and takes less than a minute. Read more

Click here to reset your password.
Sign in to get notified via email when new comments are made.