Sun Shih-Hung, Lee Mon-Juan, Lee Yun-Han, Lee Wei, Song Xiaolong, Chen Chao-Yuan
Institute of Imaging and Biomedical Photonics, College of Photonics, National Chiao Tung University, Guiren Dist., Tainan 71150, Taiwan.
Department of Bioscience Technology, Chang Jung Christian University, Guiren Dist., Tainan 71101, Taiwan ;
Biomed Opt Express. 2014 Dec 23;6(1):245-56. doi: 10.1364/BOE.6.000245. eCollection 2015 Jan 1.
The use of fluorescence is ubiquitously found in the detection of immunoreaction; though with good sensitivity, this technique requires labeling as well as other time-consuming steps to perform the measurement. An alternative approach involving liquid crystals (LCs) was proposed, based on the fact that an immunocomplex can disturb the orientation of LCs, leading to an optical texture different from the case when only antigen or antibody exists. This method is label-free, easy to manipulate and low-cost. However, its sensitivity was low for practical usage. In this study, we adopted a high-birefringence liquid crystal (LC) to enhance the sensitivity for the immunodetection. Experiments were performed, targeting at the cancer biomarker CA125. We showed that the larger birefringence (Δn = 0.33 at 20 °C) amplifies the detected signal and, in turn, dramatically improves the detection limit. To avoid signal loss from conventional rinsing steps in immunodetection, CA125 antigen and antibody were reacted before immobilized on substrates. We studied the specific binding events and obtained a detection limit as low as 1 ng/ml. The valid temperature ranges were compared by using the typical single-compound LC 5CB and the high-birefringence LC mixture. We further investigated time dependency of the optical textures and affirmed the capability of LC-based immunodetection in distinguishing between specific and nonspecific antibodies.
荧光技术在免疫反应检测中广泛应用;尽管该技术灵敏度高,但需要标记以及其他耗时步骤来进行测量。基于免疫复合物会干扰液晶(LC)取向这一事实,人们提出了一种涉及液晶的替代方法,这会导致光学纹理与仅存在抗原或抗体时不同。该方法无需标记、易于操作且成本低。然而,其灵敏度在实际应用中较低。在本研究中,我们采用高双折射液晶来提高免疫检测的灵敏度。针对癌症生物标志物CA125进行了实验。我们发现,更大的双折射(20℃时Δn = 0.33)放大了检测信号,进而显著提高了检测限。为避免免疫检测中传统冲洗步骤导致的信号损失,在将CA125抗原和抗体固定在底物上之前使其发生反应。我们研究了特异性结合事件,获得了低至1 ng/ml的检测限。通过使用典型的单化合物液晶5CB和高双折射液晶混合物比较了有效温度范围。我们进一步研究了光学纹理的时间依赖性,并确认了基于液晶的免疫检测区分特异性和非特异性抗体的能力。