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Improved application of the electrophoretic tissue clearing technology, CLARITY, to intact solid organs including brain, pancreas, liver, kidney, lung, and intestine.

Lee H, Park JH, Seo I, Park SH, Kim S - BMC Dev. Biol. (2014)

Bottom Line: We determined the optimal conditions for reducing bubble formation, discoloration, and depositing of black particles on the surface of tissue, which allowed production of clearer organ images.We developed improved CLARITY methods for clearing of the brain, pancreas, lung, intestine, liver, and kidney, and identified the appropriate experimental conditions for clearing of each specific tissue type.These optimized methods will be useful for the application of CLARITY to various types of organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Keimyung University School of Medicine, 1095 Dalgubeoldae-Ro, Dalseo-Gu, Daegu, 704-701, South Korea. neuroana@dsmc.or.kr.

ABSTRACT

Background: Mapping of tissue structure at the cellular, circuit, and organ-wide scale is important for understanding physiological and biological functions. A bio-electrochemical technique known as CLARITY used for three-dimensional anatomical and phenotypical mapping within transparent intact tissues has been recently developed. This method provided a major advance in understanding the structure-function relationships in circuits of the nervous system and organs by using whole-body clearing. Thus, in the present study, we aimed to improve the original CLARITY procedure and developed specific CLARITY protocols for various intact organs.

Results: We determined the optimal conditions for reducing bubble formation, discoloration, and depositing of black particles on the surface of tissue, which allowed production of clearer organ images. We also determined the appropriate replacement cycles of clearing solution for each type of organ, and convincingly demonstrated that 250-280 mA is the ideal range of electrical current for tissue clearing. We then acquired each type of cleared organs including brain, pancreas, liver, lung, kidney, and intestine. Additionally, we determined the images of axon fibers of hippocampal region, the Purkinje layer of cerebellum, and vessels and cellular nuclei of pancreas.

Conclusions: CLARITY is an innovative biochemical technology for the structural and molecular analysis of various types of tissue. We developed improved CLARITY methods for clearing of the brain, pancreas, lung, intestine, liver, and kidney, and identified the appropriate experimental conditions for clearing of each specific tissue type. These optimized methods will be useful for the application of CLARITY to various types of organs.

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CLARITY of intact adult mouse tissues. In adult mouse tissues (12 weeks old), imaging was performed before and/or after CLARITY. (A) Brain. (B) Pancreas. (C) Lung. (D) Intestine. (E) Liver. (F) Kidney.
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Fig1: CLARITY of intact adult mouse tissues. In adult mouse tissues (12 weeks old), imaging was performed before and/or after CLARITY. (A) Brain. (B) Pancreas. (C) Lung. (D) Intestine. (E) Liver. (F) Kidney.

Mentions: We found that for brain samples, 280 mA was the optimal current without causing any damage. When the constant current of 280 mA was maintained across the sample and the clearing solution was replaced three times over 2 weeks, the sample was completely cleared (Figure 1A). Additionally, it was determined that at a constant electrical current condition, the measured voltage between the platinum wires of chamber should reach at least 20–30 V for successful tissue clearing.Figure 1


Improved application of the electrophoretic tissue clearing technology, CLARITY, to intact solid organs including brain, pancreas, liver, kidney, lung, and intestine.

Lee H, Park JH, Seo I, Park SH, Kim S - BMC Dev. Biol. (2014)

CLARITY of intact adult mouse tissues. In adult mouse tissues (12 weeks old), imaging was performed before and/or after CLARITY. (A) Brain. (B) Pancreas. (C) Lung. (D) Intestine. (E) Liver. (F) Kidney.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4281481&req=5

Fig1: CLARITY of intact adult mouse tissues. In adult mouse tissues (12 weeks old), imaging was performed before and/or after CLARITY. (A) Brain. (B) Pancreas. (C) Lung. (D) Intestine. (E) Liver. (F) Kidney.
Mentions: We found that for brain samples, 280 mA was the optimal current without causing any damage. When the constant current of 280 mA was maintained across the sample and the clearing solution was replaced three times over 2 weeks, the sample was completely cleared (Figure 1A). Additionally, it was determined that at a constant electrical current condition, the measured voltage between the platinum wires of chamber should reach at least 20–30 V for successful tissue clearing.Figure 1

Bottom Line: We determined the optimal conditions for reducing bubble formation, discoloration, and depositing of black particles on the surface of tissue, which allowed production of clearer organ images.We developed improved CLARITY methods for clearing of the brain, pancreas, lung, intestine, liver, and kidney, and identified the appropriate experimental conditions for clearing of each specific tissue type.These optimized methods will be useful for the application of CLARITY to various types of organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Keimyung University School of Medicine, 1095 Dalgubeoldae-Ro, Dalseo-Gu, Daegu, 704-701, South Korea. neuroana@dsmc.or.kr.

ABSTRACT

Background: Mapping of tissue structure at the cellular, circuit, and organ-wide scale is important for understanding physiological and biological functions. A bio-electrochemical technique known as CLARITY used for three-dimensional anatomical and phenotypical mapping within transparent intact tissues has been recently developed. This method provided a major advance in understanding the structure-function relationships in circuits of the nervous system and organs by using whole-body clearing. Thus, in the present study, we aimed to improve the original CLARITY procedure and developed specific CLARITY protocols for various intact organs.

Results: We determined the optimal conditions for reducing bubble formation, discoloration, and depositing of black particles on the surface of tissue, which allowed production of clearer organ images. We also determined the appropriate replacement cycles of clearing solution for each type of organ, and convincingly demonstrated that 250-280 mA is the ideal range of electrical current for tissue clearing. We then acquired each type of cleared organs including brain, pancreas, liver, lung, kidney, and intestine. Additionally, we determined the images of axon fibers of hippocampal region, the Purkinje layer of cerebellum, and vessels and cellular nuclei of pancreas.

Conclusions: CLARITY is an innovative biochemical technology for the structural and molecular analysis of various types of tissue. We developed improved CLARITY methods for clearing of the brain, pancreas, lung, intestine, liver, and kidney, and identified the appropriate experimental conditions for clearing of each specific tissue type. These optimized methods will be useful for the application of CLARITY to various types of organs.

Show MeSH