Histopathological Analysis of Mouse and Human Tissues

Mouse and human tissues were fixed with a 10 % formalin solution and embedded in paraffin. 4 μm sections were cut from the tissues and processed for H&E staining and immunohistochemistry using standard protocols as previously described [12 (link),16 (link)]. For immunohistochemistry (IHC) the following antibodies were used: CDA (ab82346, Abcam), DCK (ab96599, Abcam), NT5C1A (Assay Biotechnology Company Inc., C15296), Podoplanin (Axxora LLC; CVL-MAB50714), HA (385911-50UG, Merck Millipore), TYMS (#9045, Cell Signaling). All antibodies were diluted 1:200 in 1 % BSA in TBST. All slides were analyzed by using Fiji imaging software (v 1.52p and 2.14.0/1.54f) as published earlier by Schindelin et al [17 (link)]. For IHC stainings of CDA, DCK, NT5C1A and TYMS, 10 pictures were taken per slide. For stainings of stromal components (HA, Masson's trichrome, pisosirius and podoplanin) 7 pictures were taken per slide using Olympus DP27 camera and the Olympus CellSens Entry 1.12 software.

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Knoll L., Hamm J., Stroebel P., Jovan T., Goetze R., Singh S., Hessmann E., Ellenrieder V., Ammer-Herrmenau C, & Neesse A. (2024). Expression of gemcitabine metabolizing enzymes and stromal components reveal complexities of preclinical pancreatic cancer models for therapeutic testing. Neoplasia (New York, N.Y.), 53, 101002.

Publication 2024

ab82346 ab96599 DP27 camera CellSens Entry 1.12 software

Corresponding Organization : University of Göttingen

Other organizations : Medizinische Hochschule Hannover

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Variable analysis

independent variables

Tissue fixation with 10% formalin solution
Embedding tissues in paraffin

dependent variables

H&E staining
Immunohistochemistry staining using CDA, DCK, NT5C1A, Podoplanin, HA, and TYMS antibodies

control variables

Mouse and human tissues
4 μm sections of tissues
Standard protocols for H&E staining and immunohistochemistry as previously described [12, 16]
Antibody dilutions (1:200 in 1% BSA in TBST)
Imaging and analysis using Fiji imaging software (v 1.52p and 2.14.0/1.54f) as published earlier by Schindelin et al [17]

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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