Electrical Engineering Department, University of California Los Angeles, Los Angeles, California, United States of America.
PLoS One. 2011 Jan 6;6(1):e15955. doi: 10.1371/journal.pone.0015955.
We demonstrate lensfree on-chip fluorescent imaging of transgenic Caenorhabditis elegans (C. elegans) over an ultra-wide field-of-view (FOV) of e.g., >2-8 cm(2) with a spatial resolution of ∼10 µm. This is the first time that a lensfree on-chip platform has successfully imaged fluorescent C. elegans samples. In our wide-field lensfree imaging platform, the transgenic samples are excited using a prism interface from the side, where the pump light is rejected through total internal reflection occurring at the bottom facet of the substrate. The emitted fluorescent signal from C. elegans samples is then recorded on a large area opto-electronic sensor-array over an FOV of e.g., >2-8 cm(2), without the use of any lenses, thin-film interference filters or mechanical scanners. Because fluorescent emission rapidly diverges, such lensfree fluorescent images recorded on a chip look blurred due to broad point-spread-function of our platform. To combat this resolution challenge, we use a compressive sampling algorithm to uniquely decode the recorded lensfree fluorescent patterns into higher resolution images, demonstrating ∼10 µm resolution. We tested the efficacy of this compressive decoding approach with different types of opto-electronic sensors to achieve a similar resolution level, independent of the imaging chip. We further demonstrate that this wide FOV lensfree fluorescent imaging platform can also perform sequential bright-field imaging of the same samples using partially-coherent lensfree digital in-line holography that is coupled from the top facet of the same prism used in fluorescent excitation. This unique combination permits ultra-wide field dual-mode imaging of C. elegans on a chip which could especially provide a useful tool for high-throughput screening applications in biomedical research.
我们展示了无透镜片上荧光成像技术,能够在超宽视场(例如,>2-8cm2)下对转基因秀丽隐杆线虫(Caenorhabditis elegans,C. elegans)进行成像,空间分辨率约为 10µm。这是首次成功对荧光 C. elegans 样本进行无透镜片上成像。在我们的宽场无透镜成像平台中,通过棱镜接口从侧面激发转基因样本,其中泵浦光通过在基底下表面发生全内反射被拒绝。然后,通过使用大面积光电传感器阵列记录来自 C. elegans 样本的发射荧光信号,视场范围为例如,>2-8cm2,无需使用任何透镜、薄膜干涉滤光片或机械扫描仪。由于荧光发射迅速发散,因此由于我们平台的宽点扩散函数,在芯片上记录的无透镜荧光图像看起来模糊。为了解决这个分辨率挑战,我们使用压缩采样算法将记录的无透镜荧光模式唯一地解码为更高分辨率的图像,展示了约 10µm 的分辨率。我们使用不同类型的光电传感器测试了这种压缩解码方法的有效性,以实现类似的分辨率水平,而与成像芯片无关。我们进一步证明,这种宽视场无透镜荧光成像平台还可以使用部分相干无透镜数字线全息术从用于荧光激发的相同棱镜的上表面耦合,对相同的样本进行顺序明场成像。这种独特的组合允许在芯片上对 C. elegans 进行超宽场双模成像,这特别为生物医学研究中的高通量筛选应用提供了有用的工具。