Patel Yogesh G, Rajadhyaksha Milind, Dimarzio Charles A
Biomed Opt Express. 2011 Aug 1;2(8):2231-42. doi: 10.1364/BOE.2.002231. Epub 2011 Jul 8.
Both point-scanning and line-scanning confocal microscopes provide resolution and optical sectioning to observe nuclear and cellular detail in human tissues, and are being translated for clinical applications. While traditional point-scanning is truly confocal and offers the best possible optical sectioning and resolution, line-scanning is partially confocal but may offer a relatively simpler and lower-cost alternative for more widespread dissemination into clinical settings. The loss of sectioning and loss of contrast due to scattering in tissue is more rapid and more severe with a line-scan than with a point-scan. However, the sectioning and contrast may be recovered with the use of a divided-pupil. Thus, as part of our efforts to translate confocal microscopy for detection of skin cancer, and to determine the best possible approach for clinical applications, we are now developing a quantitative understanding of imaging performance for a set of scanning and pupil conditions. We report a Fourier-analysis-based computational model of confocal microscopy for six configurations. The six configurations are point-scanning and line-scanning, with full-pupil, half-pupil and divided-pupils. The performance, in terms of on-axis irradiance (signal), resolution and sectioning capabilities, is quantified and compared among these six configurations.
点扫描共聚焦显微镜和线扫描共聚焦显微镜都能提供分辨率和光学切片功能,用于观察人体组织中的细胞核和细胞细节,并且正在转化为临床应用。传统的点扫描是真正的共聚焦,能提供最佳的光学切片和分辨率,而线扫描是部分共聚焦,但可能为更广泛地应用于临床提供一种相对简单且成本较低的替代方案。与点扫描相比,线扫描时由于组织散射导致的切片损失和对比度损失更快、更严重。然而,使用分瞳技术可以恢复切片和对比度。因此,作为我们将共聚焦显微镜转化用于皮肤癌检测以及确定临床应用最佳方法的努力的一部分,我们目前正在对一组扫描和光瞳条件下的成像性能进行定量理解。我们报告了基于傅里叶分析的共聚焦显微镜六种配置的计算模型。这六种配置分别是点扫描和线扫描,以及全光瞳、半光瞳和分瞳。对这六种配置在轴上辐照度(信号)、分辨率和切片能力方面的性能进行了量化和比较。