Ishmukhametov Robert R, Russell Aidan N, Wheeler Richard J, Nord Ashley L, Berry Richard M
Department of Physics, Clarendon Laboratory, University of Oxford, Parks Road, Oxford, OX1 3PU, UK.
The Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK.
Sci Rep. 2016 Feb 8;6:20729. doi: 10.1038/srep20729.
Back-scattering darkfield (BSDF), epi-fluorescence (EF), interference reflection contrast (IRC), and darkfield surface reflection (DFSR) are advanced but expensive light microscopy techniques with limited availability. Here we show a simple optical design that combines these four techniques in a simple low-cost miniature epi-illuminator, which inserts into the differential interference-contrast (DIC) slider bay of a commercial microscope, without further additions required. We demonstrate with this device: 1) BSDF-based detection of Malarial parasites inside unstained human erythrocytes; 2) EF imaging with and without dichroic components, including detection of DAPI-stained Leishmania parasite without using excitation or emission filters; 3) RIC of black lipid membranes and other thin films, and 4) DFSR of patterned opaque and transparent surfaces. We believe that our design can expand the functionality of commercial bright field microscopes, provide easy field detection of parasites and be of interest to many users of light microscopy.
背散射暗场(BSDF)、落射荧光(EF)、干涉反射对比度(IRC)和暗场表面反射(DFSR)是先进但昂贵的光学显微镜技术,且可用性有限。在此,我们展示了一种简单的光学设计,该设计将这四种技术集成在一个简单的低成本微型落射照明器中,该照明器可插入商用显微镜的微分干涉对比(DIC)滑块槽中,无需进一步添加其他部件。我们用该设备演示了:1)基于BSDF检测未染色人红细胞内的疟原虫;2)有无二向色性组件的EF成像,包括不使用激发或发射滤光片检测经4',6-二脒基-2-苯基吲哚(DAPI)染色的利什曼原虫寄生虫;3)黑色脂质膜和其他薄膜的IRC成像,以及4)图案化不透明和透明表面的DFSR成像。我们相信,我们的设计可以扩展商用明场显微镜的功能,便于现场检测寄生虫,并且会引起许多光学显微镜用户的兴趣。