A novel dual-channel single-polarization photonic crystal fiber (PCF) sensor is proposed for multi-analyte detection. The sensor is fabricated using the stack-and-draw technique and demonstrates high sensitivity with maximum wavelength and amplitude sensitivities of 11,000 nm/RIU and 807 RIU⁻¹, respectively. The sensor operates in single polarization mode, offering lower confinement loss and better wavelength resolution. It is capable of detecting multiple analytes simultaneously, with each channel responding to different refractive indices. The sensor's performance is evaluated through numerical simulations and experimental testing, showing high sensitivity and minimal false positives. The sensor's design allows for the detection of a wide range of analyte refractive indices, from 1.33 to 1.41, including water, milk, acetic acid, decane, and breast cancer cells. The sensor's performance is further enhanced by optimizing the gold layer thickness, achieving a maximum figure of merit (FOM) of 204 RIU⁻¹. The proposed sensor outperforms existing multi-analyte detection sensors due to its high sensitivity, dual-channel configuration, and superior sensing capabilities. It has potential applications in disease detection, such as cancer, diarrhea, and malaria, as well as industrial chemical analysis. The sensor's ability to detect a broad range of analyte refractive indices makes it suitable for various sensing applications.A novel dual-channel single-polarization photonic crystal fiber (PCF) sensor is proposed for multi-analyte detection. The sensor is fabricated using the stack-and-draw technique and demonstrates high sensitivity with maximum wavelength and amplitude sensitivities of 11,000 nm/RIU and 807 RIU⁻¹, respectively. The sensor operates in single polarization mode, offering lower confinement loss and better wavelength resolution. It is capable of detecting multiple analytes simultaneously, with each channel responding to different refractive indices. The sensor's performance is evaluated through numerical simulations and experimental testing, showing high sensitivity and minimal false positives. The sensor's design allows for the detection of a wide range of analyte refractive indices, from 1.33 to 1.41, including water, milk, acetic acid, decane, and breast cancer cells. The sensor's performance is further enhanced by optimizing the gold layer thickness, achieving a maximum figure of merit (FOM) of 204 RIU⁻¹. The proposed sensor outperforms existing multi-analyte detection sensors due to its high sensitivity, dual-channel configuration, and superior sensing capabilities. It has potential applications in disease detection, such as cancer, diarrhea, and malaria, as well as industrial chemical analysis. The sensor's ability to detect a broad range of analyte refractive indices makes it suitable for various sensing applications.