Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, PR China.
Appl Spectrosc. 2010 Aug;64(8):847-52. doi: 10.1366/000370210792081154.
The differing bioconjugation efficiencies of quantum dots (QDs) are a practical obstacle to their popularization. Differences in bioconjugation efficiency based on immunostaining the same targeted molecules using different batches of QDs need to be evaluated prior to their application. In this paper, a quantitative method for evaluating the efficiency of QDs in staining tissues has been developed based on Hadamard transform (HT) spectral imaging. Proliferating cell nuclear antigens (PCNA) in breast cancer tissues were labeled with bioconjugated QD bioprobes using a 454 nm laser as the light source for fluorescence spectral imaging. Four-dimensional (4D) spectral imaging analysis of PCNA in cell nuclei was carried out using HT spectral microscopy based on immunostaining with different batches of QDs. The fluorescence intensity distributions in the cell nuclei were collected from the 4D images. Based on the information obtained from microscopic spectra and 4D images, differences in the bioconjugation efficiency among different batches of QDs were evaluated. The results demonstrate that it is possible to maintain uniform bioconjugation efficiencies with different QD bioconjugation processes in order to obtain accurate and reliable results in biomedical analysis and cancer diagnosis.
量子点（QD）的生物偶联效率差异是其推广的一个实际障碍。在将相同的靶向分子用不同批次的 QD 进行免疫染色之前，需要评估基于免疫染色的生物偶联效率差异。本文基于 Hadamard 变换（HT）光谱成像开发了一种定量评估 QD 染色组织效率的方法。使用 454nm 激光作为荧光光谱成像的光源，用生物共轭 QD 生物探针标记乳腺癌组织中的增殖细胞核抗原（PCNA）。基于用不同批次的 QD 进行免疫染色的 HT 光谱显微镜，对细胞核内的 PCNA 进行了四维（4D）光谱成像分析。从 4D 图像中收集细胞核内的荧光强度分布。基于从微观光谱和 4D 图像中获得的信息，评估了不同批次 QD 之间的生物偶联效率差异。结果表明，通过不同的 QD 生物偶联过程保持均匀的生物偶联效率是可能的，从而在生物医学分析和癌症诊断中获得准确可靠的结果。
