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( A ) Fluorescence images demonstrated the differences between the IR-780–labeled proteins in 4T1 tumor, breast, and muscle lysates following the separation using the 2D SDS-PAGE. ( B ) Western blotting for detecting overexpressed albumin and <t>AKT1</t> in the 4T1 tumor sample. ( C ) Fraction 1 was selected to distinguish tumor and muscle tissues. Quantification of the Fraction 1–to–Fraction 2 (actin) ratio as annotated in (A), albumin-to-actin ratio, and AKT1-to-actin ratio as annotated in (A) (** P < 0.05). ( D ) IHC staining of albumin and AKT1 levels in 4T1 tumor and muscle tissues. Scale bar, 50 μm. ( E ) Western blotting for detecting albumin and AKT1 levels in a set of 4T1 tumor samples collected after 3, 7, 21, and 35 days of inoculation, respectively. ( F ) Schematic representation of the workflow of the 3D histological electrophoresis for tissue sections. ( G ) Two samples with either muscle&muscle or 4T1 tumor&muscle combination were subjected to the 3D histological electrophoresis. ( H ) The tumor-to-muscle ratios were analyzed and plotted before/after the separation by 3D histological electrophoresis ( n = 3 mice per group, **** P < 0.05). ( I ) By collecting a set of 4T1 tumor samples after 3, 7, 21, and 35 days of inoculation, the tumor-to-muscle ratios were analyzed and plotted after the 3D histological electrophoresis ( n = 3 mice per group). ( J ) NIR images and signal quantification of the seven fractionated layers for the muscle&muscle sample after the 3D histological electrophoresis. Scale bar, 1 cm. ( K ) NIR images and signal quantification of seven fractionated layers for the tumor&muscle sample following the 3D histological electrophoresis. Layers 2, 3, and 4 were observed with the maximum signal difference between tumors and muscles, primed for assessing the tumor-positive margins. Scale bar, 1 cm.
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( A ) Fluorescence images demonstrated the differences between the IR-780–labeled proteins in 4T1 tumor, breast, and muscle lysates following the separation using the 2D SDS-PAGE. ( B ) Western blotting for detecting overexpressed albumin and AKT1 in the 4T1 tumor sample. ( C ) Fraction 1 was selected to distinguish tumor and muscle tissues. Quantification of the Fraction 1–to–Fraction 2 (actin) ratio as annotated in (A), albumin-to-actin ratio, and AKT1-to-actin ratio as annotated in (A) (** P < 0.05). ( D ) IHC staining of albumin and AKT1 levels in 4T1 tumor and muscle tissues. Scale bar, 50 μm. ( E ) Western blotting for detecting albumin and AKT1 levels in a set of 4T1 tumor samples collected after 3, 7, 21, and 35 days of inoculation, respectively. ( F ) Schematic representation of the workflow of the 3D histological electrophoresis for tissue sections. ( G ) Two samples with either muscle&muscle or 4T1 tumor&muscle combination were subjected to the 3D histological electrophoresis. ( H ) The tumor-to-muscle ratios were analyzed and plotted before/after the separation by 3D histological electrophoresis ( n = 3 mice per group, **** P < 0.05). ( I ) By collecting a set of 4T1 tumor samples after 3, 7, 21, and 35 days of inoculation, the tumor-to-muscle ratios were analyzed and plotted after the 3D histological electrophoresis ( n = 3 mice per group). ( J ) NIR images and signal quantification of the seven fractionated layers for the muscle&muscle sample after the 3D histological electrophoresis. Scale bar, 1 cm. ( K ) NIR images and signal quantification of seven fractionated layers for the tumor&muscle sample following the 3D histological electrophoresis. Layers 2, 3, and 4 were observed with the maximum signal difference between tumors and muscles, primed for assessing the tumor-positive margins. Scale bar, 1 cm.

Journal: Science Advances

Article Title: Three-dimensional histological electrophoresis enables fast automatic distinguishment of cancer margins and lymph node metastases

doi: 10.1126/sciadv.adg2690

Figure Lengend Snippet: ( A ) Fluorescence images demonstrated the differences between the IR-780–labeled proteins in 4T1 tumor, breast, and muscle lysates following the separation using the 2D SDS-PAGE. ( B ) Western blotting for detecting overexpressed albumin and AKT1 in the 4T1 tumor sample. ( C ) Fraction 1 was selected to distinguish tumor and muscle tissues. Quantification of the Fraction 1–to–Fraction 2 (actin) ratio as annotated in (A), albumin-to-actin ratio, and AKT1-to-actin ratio as annotated in (A) (** P < 0.05). ( D ) IHC staining of albumin and AKT1 levels in 4T1 tumor and muscle tissues. Scale bar, 50 μm. ( E ) Western blotting for detecting albumin and AKT1 levels in a set of 4T1 tumor samples collected after 3, 7, 21, and 35 days of inoculation, respectively. ( F ) Schematic representation of the workflow of the 3D histological electrophoresis for tissue sections. ( G ) Two samples with either muscle&muscle or 4T1 tumor&muscle combination were subjected to the 3D histological electrophoresis. ( H ) The tumor-to-muscle ratios were analyzed and plotted before/after the separation by 3D histological electrophoresis ( n = 3 mice per group, **** P < 0.05). ( I ) By collecting a set of 4T1 tumor samples after 3, 7, 21, and 35 days of inoculation, the tumor-to-muscle ratios were analyzed and plotted after the 3D histological electrophoresis ( n = 3 mice per group). ( J ) NIR images and signal quantification of the seven fractionated layers for the muscle&muscle sample after the 3D histological electrophoresis. Scale bar, 1 cm. ( K ) NIR images and signal quantification of seven fractionated layers for the tumor&muscle sample following the 3D histological electrophoresis. Layers 2, 3, and 4 were observed with the maximum signal difference between tumors and muscles, primed for assessing the tumor-positive margins. Scale bar, 1 cm.

Article Snippet: AKT1 was purchased from MedChemExpress.

Techniques: Fluorescence, Labeling, SDS Page, Western Blot, Immunohistochemistry, Electrophoresis, Muscles