Department of Precision Instrument, State Key Laboratory of Precision Measurement Technology and Instruments, Tsinghua University, Beijing 100084, China.
Department of Chemistry, Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084, China.
Anal Chem. 2021 Jul 27;93(29):10056-10064. doi: 10.1021/acs.analchem.1c00660. Epub 2021 Jul 12.
Existing structured illumination microscopy (SIM) allows super-resolution live-cell imaging in few color channels that provide merely morphological information but cannot acquire the sample spectrum that is strongly relevant to the underlying physicochemical property. We develop hyperspectral SIM which enables high-speed spectral super-resolution imaging in SIM for the first time. Through optically mapping the three-dimensional (, , and λ) datacube of the sample to the detector plane, hyperspectral SIM allows snapshot spectral imaging of the SIM raw image, detecting the sample spectrum while retaining the high-speed and super-resolution characteristics of SIM. We demonstrate hyperspectral SIM imaging and reconstruct a datacube containing 31 super-resolution images of different wavelengths from only 9 exposures, achieving a 15 nm spectral resolution. We show time-lapse hyperspectral SIM imaging that achieves an imaging speed of 2.7 s per datacube-31-fold faster than the existing wavelength scanning strategy. To demonstrate the great prospects for further combining hyperspectral SIM with various spectral analysis methods, we also perform spectral unmixing of the hyperspectral SIM result while imaging the spectrally overlapped sample.
现有的结构光照明显微镜（SIM）允许在少数颜色通道中进行超分辨率活细胞成像，这些颜色通道仅提供形态信息，但无法获取与潜在物理化学性质强相关的样本光谱。我们开发了超高光谱 SIM，这是首次在 SIM 中实现高速光谱超分辨率成像。通过将样品的三维（，，和 λ）数据立方体光学映射到探测器平面，超高光谱 SIM 允许对 SIM 原始图像进行快照光谱成像，在保留 SIM 的高速和超分辨率特性的同时检测样品光谱。我们演示了超高光谱 SIM 成像，并从仅 9 次曝光中重建了一个包含 31 个不同波长的超分辨率图像的数据立方体，实现了 15nm 的光谱分辨率。我们展示了时移超高光谱 SIM 成像，其成像速度为每数据立方体 2.7 秒，比现有的波长扫描策略快 31 倍。为了展示将超高光谱 SIM 与各种光谱分析方法进一步结合的广阔前景，我们还在对光谱重叠样本进行成像的同时，对超高光谱 SIM 的结果进行光谱解混。
