Liver wedge biopsies were taken during the back-table preparation. All biopsy samples were placed in HTK solution (Custodiol®, Dr. Franz Köhler Chemie GmbH, Bensheim, Germany) on ice for transportation prior to analysis.
Real-time live confocal microscopy assessment was performed using the following live stains: Wheat germ agglutinin conjugate (WGA; Molecular Probes, Eugene, OR, United States; 10 μg·mL−1 final concentration) visualizes the tissue morphology, SYTO®16 (Molecular Probes; final concentration 5 µM) visualizes all nuclei and propidium iodide (PI) (Molecular Probes; final concentration 500 nM) the nuclei of dead cells [20 (link)]. Incubation time was 15 min at 37°C. Real-time live confocal imaging was performed in eight-well chambered cover glasses (Nalge Nunc International). Images were acquired with a spinning disk confocal system (UltraVIEW VoX; Perkin Elmer, Waltham, MA) connected to a Zeiss Axio Observer Z1 microscope (Zeiss, Oberkochen, Germany) and visualized employing the Volocity software (Perkin Elmer) using a ×10 objective. Time for readout was approximately 5 min per sample.
Real-time live confocal microscopy assessment was performed using the following live stains: Wheat germ agglutinin conjugate (WGA; Molecular Probes, Eugene, OR, United States; 10 μg·mL−1 final concentration) visualizes the tissue morphology, SYTO®16 (Molecular Probes; final concentration 5 µM) visualizes all nuclei and propidium iodide (PI) (Molecular Probes; final concentration 500 nM) the nuclei of dead cells [20 (link)]. Incubation time was 15 min at 37°C. Real-time live confocal imaging was performed in eight-well chambered cover glasses (Nalge Nunc International). Images were acquired with a spinning disk confocal system (UltraVIEW VoX; Perkin Elmer, Waltham, MA) connected to a Zeiss Axio Observer Z1 microscope (Zeiss, Oberkochen, Germany) and visualized employing the Volocity software (Perkin Elmer) using a ×10 objective. Time for readout was approximately 5 min per sample.
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