This study investigated the combination of two contrast agents for ex vivo cancer detection in breast tissues. Samples were stained with molecular marker pH Low Insertion Peptide (pHLIP) conjugated with fluorescent dye Alexa532 (pHLIP–Alexa532), and intravital stain Methylene Blue (MB), and imaged with a high-resolution confocal microscope. Resulting images display cellular morphology with staining patterns mimicking Hematoxylin and Eosin histopathology and show promise for the method used as a tool for breast cancer detection in biopsies or intraoperatively.
Quick, macroscopic examination of breast tissue from surgical excisions or biopsies can guide breast cancer patient management. To this purpose, we examined dual-contrast fluorescence imaging with intravital dye Methylene Blue (MB) and disease-specific molecular marker pH low insertion peptide (pHLIP) conjugated with fluorescent Alexa532 (Alexa532-pHLIP) as contrast agents. Samples were stained with both MB and Alexa532-pHLIP and imaged with multimodal wide-field system. Co- and cross-polarized reflectance and fluorescence images were acquired. Then, specimens were processed for H and E paraffin embedded histopathology. Wide-field imaging demonstrated increased pHLIP-Alexa 532 fluorescence emission and high MB fluorescence polarization in cancerous regions.
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