Pan An, Zhang Yan, Wen Kai, Zhou Meiling, Min Junwei, Lei Ming, Yao Baoli
Opt Express. 2018 Sep 3;26(18):23119-23131. doi: 10.1364/OE.26.023119.
Fourier ptychography (FP) is a promising computational imaging technique that overcomes the physical space-bandwidth product (SBP) limit of a conventional microscope by applying angular-varied illuminations. However, to date, the effective imaging numerical aperture (NA) achievable with a commercial LED board is still limited to the range of 0.3-0.7 with a 4 × /0.1NA objective due to the geometric constraint with the declined illumination intensities and attenuated signal-to-noise ratio (SNR). Thus the highest achievable half-pitch resolution is usually constrained between 500-1000 nm, which cannot meet the requirements of high-resolution biomedical imaging applications. Although it is possible to improve the resolution by using a high-NA objective lens, the FP approach is less appealing as the decrease of field-of-view (FOV) will far exceed the improvement of spatial resolution in this case. In this paper, we initially present a subwavelength resolution Fourier ptychography (SRFP) platform with a hemispherical digital condenser to provide high-angle programmable plane-wave illuminations of 0.95NA, attaining a 4 × /0.1NA objective with the final effective imaging performance of 1.05NA at a half-pitch resolution of 244 nm with the incident wavelength of 465 nm across a wide FOV of 14.60 mm, corresponding to a SBP of 245 megapixels. Our work provides an essential step of FP towards high-throughput imaging applications.
傅里叶叠层成像术(FP)是一种很有前景的计算成像技术,它通过应用角度变化的照明来克服传统显微镜的物理空间带宽积(SBP)限制。然而,到目前为止,由于照明强度下降和信噪比(SNR)衰减带来的几何约束,使用商用LED板可实现的有效成像数值孔径(NA)在搭配4×/0.1NA物镜时仍限于0.3 - 0.7的范围。因此,可实现的最高半间距分辨率通常限制在500 - 1000 nm之间,这无法满足高分辨率生物医学成像应用的要求。虽然使用高NA物镜有可能提高分辨率,但在这种情况下,由于视场(FOV)的减小将远远超过空间分辨率的提高,FP方法的吸引力较小。在本文中,我们首先展示了一个具有半球形数字聚光镜的亚波长分辨率傅里叶叠层成像术(SRFP)平台,以提供0.95NA的高角度可编程平面波照明,在465 nm入射波长下,搭配4×/0.1NA物镜,在14.60 mm的宽视场内实现了半间距分辨率为244 nm时最终有效成像性能为1.05NA,对应245兆像素的SBP。我们的工作为FP迈向高通量成像应用迈出了重要一步。