The 33rd International Congress on High-Speed Imaging and Photonics grandly opened in Shenzhen on the 16th, gathering the latest progress of global cutting-edge imaging technologies. The previous issue introduced the applications of sCMOS cameras in X-ray scintillator imaging, computational imaging, and high-energy physics fields. This issue will focus on the cell and biological imaging fields in biomedicine, and analyze typical applications in SIM super-resolution imaging, single-molecule fluorescence imaging, and wide-field fluorescence imaging, combined with practical verification of the self-developed Revealer Gloria series sCMOS cameras by HF Agile Device Co.,Ltd.
Traditional optical microscopes are limited by the diffraction limit, making it difficult to clearly present fine cell structures at the 100nm scale. Structured Illumination Microscopy (SIM) is a wide-field super-resolution imaging technology. Through spatially modulated illumination and frequency-domain reconstruction algorithms, it can improve the spatial resolution to ~100nm, and is applied to high-precision observation of subcellular structures such as cytoskeletons and mitochondrial networks.
SIM imaging relies on multi-frame fringe images with different phases and directions for Fourier domain reconstruction, so it has requirements for the camera's acquisition speed, signal-to-noise ratio (SNR), and field of view. The Revealer sCMOS camera Gloria 4.2 has a 95% peak quantum efficiency (QE) and 1.2 e- readout noise, ensuring high-SNR original images under low phototoxicity. Its high-speed acquisition capability of >100 fps at 2048×2048 resolution effectively covers a larger cell area and reduces motion artifacts.
In an experiment, the Gloria 4.2 scientific camera with high quantum efficiency was integrated into a SIM super-resolution microscopy system, realizing 100nm-level resolution imaging of cell samples at 100 fps. This breaks through the diffraction limit and allows clear identification of microtubule fiber branch points, cross shapes, etc. (see Figure 1).
Figure 1
Single-molecule fluorescence imaging is a core technology for studying molecular biological processes such as protein dynamics, DNA-protein interactions, and membrane receptor diffusion. The core challenge of imaging is that the number of photons emitted by a single molecule is only at the level of 20-30 photons, and the signal is easily submerged by detector noise.
The Revealer sCMOS camera Gloria 1605 provided by HF Agile Device Co.,Ltd has reduced the readout noise to 0.9 e- through process and circuit design optimization. With the support of a real-time noise suppression algorithm, it further eliminates background interference and captures clear single-molecule fluorescence signals at the level of only 20 photons, as shown in Figure 2.
Figure 2
Wide-field fluorescence imaging is the most basic and widely used imaging modality in life science research, suitable for scenarios such as cell labeling, subcellular organelle localization, and gene expression analysis. The Revealer Gloria series sCMOS cameras have been applied to various wide-field imaging systems such as spinning disk confocal microscopes, light-sheet microscopes, and flow cytometers. Their sensitivity, balance, and stability have been verified in practice, as shown in Figure 3.
Figure 3
With high quantum efficiency, low readout noise, and a balanced combination of field of view and frame rate, Revealer sCMOS cameras are changing the technical pattern of cell and biological imaging: In SIM super-resolution imaging, the large field of view and high frame rate ensure the accuracy and efficiency of computational reconstruction; In single-molecule fluorescence imaging, low readout noise and high quantum efficiency enable the capture of extremely weak signals at the 20-photon level; In wide-field fluorescence imaging, balanced comprehensive performance supports long-term high-dynamic observation of living cells...
Revealer scientific instruments will continue to research advanced imaging technologies and provide efficient and accurate scientific imaging solutions for research in the cell and biological imaging fields.
