Irreproducibility in molecular optical sectioning microscopy has hindered the transformation of acquired digital images from qualitative descriptions to quantitative data. Although numerous tools, metrics, and phantoms have been developed, accurate quantitative comparisons of data from different microscopy systems with diverse acquisition conditions remains a challenge. Here, we develop a simple tool based on an absolute measurement of bulk fluorophore solutions with related Poisson photon statistics, to overcome this obstacle. Demonstrated in a prototypical multiphoton microscope, our tool unifies the unit of pixelated measurement to enable objective comparison of imaging performance across different modalities, microscopes, components/settings, and molecular targets. The application of this tool in live specimens identifies an attractive methodology for quantitative imaging, which rapidly acquires low signal-to-noise frames with either gentle illumination or low-concentration fluorescence labeling.
Laser & photonics reviews. 2024 Oct;18(10):2400031. doi: 10.1002/lpor.202400031 Q19.82024
Pixelation with concentration-encoded effective photons for quantitative molecular optical sectioning microscopy
具有浓度编码的有效光子的像素化定量分子光学切片显微镜 翻译改进
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DOI: 10.1002/lpor.202400031 PMID: 39781104
摘要 Ai翻译
Keywords:pixelation
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