PV was cleaned from surrounded tissue. A small incision was made in PV using spring scissor. Silk suture was loosely wrapped around the cleaned PV above the incision. Tubing (PE10) ~15 cm long connected to (27G) needle was inserted into the PV incision and secured with silk suture. Green MICROFIL was injected into the PV until blood vessels on the surface were filled or resistance was met. Massaging the liver with a cotton swab while injecting helped to introduce the MICROFIL. The tubing was removed and silk suture was tightened around the PV to prevent leakage.

Liver was dissected out and placed at 4°C overnight (ON) for MICROFIL to solidify. The next day the liver was fixed with 3.7% formaldehyde solution (FA) (Sigma-Aldrich cat. #F1635) diluted in Dulbecco's phosphate-buffered saline (DPBS) (Life Technologies cat. # 14190144). After 24 hr, liver was washed and kept in DPBS. Liver was separated into lobes. The left lateral lobe was placed in 50% methanol (Sigma-Aldrich, cat. # 322415) for 4 hr and into 100% methanol ON. Further, the lobe was placed in benzyl alcohol (Sigma-Aldrich, cat. #402834) and benzyl benzoate (Sigma-Aldrich, cat. #B6630) (BA:BB 1:2) solution until transparent. The right medial lobe (only FA fixed) was used for µCT scanning. Liver lobe images of right medial lobe (P15) and left lateral lobe (P15 and adult) were taken using a stereomicroscope Stemi 305 (Carl Zeiss Microscopy) with a PowerShot S3 IS camera (Canon) or iPhone6 connected to a LabCam adapter.

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