Quantitative Analysis of Melanoma Markers

For quantitative staining analysis, Vectra 2.0 automated quantitative tissue imaging system with Nuance and inForm software 2.2.1 (PerkinElmer) was used as described previously^{39 (link)}. The multiplexed TMA was imaged with the Vectra slide scanner, using a scanning protocol that was created based on core size and layout, as well as an acquisition of spectral library. For each slide, an 8-bit image cube from each of the TMA tissue cores was acquired and the inForm advanced image analysis software was used to segment tissues (melanoma versus others) to analyze the protein levels. Then, the target signals were quantitated after unmixing the spectral curves with InForm software. Continuous signal intensity (mean optical density per pixel) was generated for each identified tissue/cell type. Data were exported for further analysis. Quantitation for PLK1, N-cadherin, and E-cadherin was performed in S100-positive cells in melanoma tissue.

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Chhabra G., Singh C.K., Ndiaye M.A., Su S., Shirley C.A, & Ahmad N. (2024). Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma. NPJ Precision Oncology, 8, 6.

Publication 2024

Vectra 2.0

Corresponding Organization : William S. Middleton Memorial Veterans Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels of PLK1, N-cadherin, and E-cadherin

control variables

Staining protocol using Vectra 2.0 automated quantitative tissue imaging system with Nuance and inForm software 2.2.1 (PerkinElmer)
Imaging of multiplexed TMA with Vectra slide scanner
Acquisition of spectral library
Segmentation of tissues (melanoma versus others) using inForm advanced image analysis software
Quantitation of target signals after unmixing the spectral curves with InForm software
Continuous signal intensity (mean optical density per pixel) generated for each identified tissue/cell type

controls

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

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