Immunostaining of FFPE Tumor Specimens

Formalin-fixed paraffin-embedded (FFPE) tumor specimens were prepared by the standard procedures in Department of Diagnostic Pathology, Kyoto University Hospital. A board-certificated pathologist selected multiple and separate cancer lesions from a single tumor. Specimens of FFPE cancer spheroids were prepared as previously reported [14 (link)]. These specimens were sectioned at 4-μm thickness, and stained with H&E or immunostained for MLH-1 (M1, Ventana, Tucson, AZ, USA), MSH2 (G219-1129, Ventana), MSH6 (EPR3945, Abcam, Cambridge, UK), or PMS2 (EPR 3947, Ventana) followed by hematoxylin counterstaining for nuclei.

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Yamaura T., Miyoshi H., Maekawa H., Morimoto T., Yamamoto T., Kakizaki F., Higasa K., Kawada K., Matsuda F., Sakai Y, & Taketo M.M. (2018). Accurate diagnosis of mismatch repair deficiency in colorectal cancer using high-quality DNA samples from cultured stem cells. Oncotarget, 9(101), 37534-37548.

Publication 2018

G219-1129 EPR3945 EPR 3947

Cancer Diagnostic Embedded paraffin Formalin Hematoxylin Msh6 Nuclei Paraffin Pathologist Pms2 Tumor

Corresponding Organization : Kyoto University

Other organizations : Kansai Medical University

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Variable analysis

independent variables

Preparation of formalin-fixed paraffin-embedded (FFPE) tumor specimens
Selection of multiple and separate cancer lesions from a single tumor by a board-certificated pathologist
Preparation of FFPE cancer spheroid specimens

dependent variables

Immunostaining of the specimens for MLH-1, MSH2, MSH6, or PMS2
Hematoxylin counterstaining for nuclei

control variables

Sectioning of the specimens at 4-μm thickness
Staining of the specimens with H&E

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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