Superresolution Imaging with Single-Antibody Labeling.
Superresolution Imaging with Single-Antibody Labeling.
复制标题
单抗体标记的超分辨率成像。
DOI：
10.1021/acs.bioconjchem.3c00178
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发表时间：
2023
影响因子：
4.7
通讯作者：
Hu,YingS
中科院分区：
文献类型：
--
作者：
Gunasekara,Hirushi;Perera,Thilini;Anderson,Jesse;Saed,Badeia;Ramseier,Neal;Keshta,Neama;Hu,YingS
我们提出了一种通用的单分子定位显微镜技术，利用单抗体标记的延时成像。通过在亚分钟时间尺度上进行单分子成像并调整抗体浓度以创建稀疏的单分子结合，我们捕获了亚细胞靶标的抗体标记以生成超分辨率图像。单抗体标记使双目标超分辨率成像使用染料共轭的单克隆抗体和多克隆抗体。我们进一步展示了一种双色策略，以增加样品标记密度。单抗体标记铺平了一条新的途径，以评估抗体结合超分辨率成像在天然细胞环境。
We present a versatile single-molecule localization microscopy technique utilizing time-lapse imaging of single-antibody labeling. By performing single-molecule imaging in the subminute time scale and tuning the antibody concentration to create sparse single-molecule binding, we captured antibody labeling of subcellular targets to generate superresolution images. Single-antibody labeling enabled dual-target superresolution imaging using dye-conjugated monoclonal and polyclonal antibodies. We further demonstrate a dual-color strategy to increase the sample labeling density. Single-antibody labeling paves a new way to evaluate antibody binding for superresolution imaging in the native cellular environment.