Paraffin-embedded human melanoma tissue microarrays (TMA) ME2082c and ME2081 were purchased from Xi’an Alenabio (Xi’an, Shanxi, China). The appendant follow-up information was used in the statistical analysis between CHPF expression and tumor characteristics. The samples were collected from patients being completely informed. The study was approved by the Ethics Committee of Fudan University. The sections were deparaffinized, citrate antigen repaired, blocked by 3% H2O2 for 10 min at room temperature, and treated by 5% goat serum for 15 min at room temperature. Then the staining of the tissue sections by diluted primary anti-CHPF or anti-CDK1 was performed 4 °C overnight. After washing, the tissue sections were stained with DAB, and counterstained with hematoxylin, dehydrated with ethanol and mounted with resin. Images were captured using ImageScope v11 (Leica, Buffalo Grove, IL, USA). The CHPF or CDK1 expression levels in each spot on TMA was assessed by the intensity (1–4, from weak to strong) and the percentage of CHPF or CDK1 positive cells (0–100%, low to high).
Imagescope v11
Manufactured by Leica camera
Sourced in United States
ImageScope v11 is a digital imaging software designed for the analysis and management of digital microscopy images. The software provides tools for viewing, annotating, and measuring various features within digital images.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using imagescope v11
1
Immunohistochemical Analysis of CHPF and CDK1
2
Immunohistochemical Analysis of CDKL3 and RRM2
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The construction of the microarray and the following immunohistochemistry were carried out as described previously by Shanghai Biochip Company.39 Glioma and normal tissue sections from glioma patients were deparaffinized, repaired with citrate antigen, blocked, and treated with 5% goat serum for 15 minutes at room temperature. The tissue sections were stained by diluted primary anti‐CDKL3 (1:200; Bioss) or anti‐RRM2 (1:400; Abcam) at 4°C overnight. Tissue sections were then stained with DAB, counterstained with hematoxylin, and dehydrated with ethanol. Images were captured using an ImageScope v11 (Leica). All sections were scored by two independent pathologists in a blinded fashion. In particular, the intensity of positive‐staining cytoplasm or nucleus of CDKL3 was scored on a scale of 0‐3 (0, negative; 1, light brown; 2, medium brown; and 3, dark brown), which was multiplied by the corresponding value of positive percentage (1, less than 25%; 2, 25%‐50%; 3, 50%‐75%; and 4, more than 75%). The two scores of cellular cytoplasm and nucleus were added to estimate the expression level of CDKL3, which was divided into the high or low cut by the median total score 4.
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