3D quantitative tomographic microscopy of low-contrast structures

相位聚焦虚拟透镜®（也称为重叠关联成像）是一种新的显微镜方法，它消除了对硬件聚焦组件的需求，从而消除了传统透镜的成本、局限性和缺陷所带来的束缚。虚拟透镜图像重建过程由复杂的计算机程序（算法）来执行，而不是透镜。由于它提供了定量的（数值）信息，并且由于它可以提取通常存在于不透明样品中的高信息含量，因此虚拟透镜在活体细胞成像和眼科（接触）透镜制造等多种应用中具有优势。如果样本的物理结构是三维的，或者如果它很厚，则可能会导致图像质量较差。因此，本工作的目的是使用新的三维虚拟透镜算法来产生复杂结构的低对比度标本的全层析定量图像。如果成功的话，预计这种新方法将能够从这样的标本中产生高质量和准确的信息，而不使用染色剂。并且可以从比传统的共焦显微镜更深的地方观察到。此外，该技术有望提供一种新的方法来精确测量，为了保证制造质量，我们对CD和DVD播放机、移动的电话、显微镜、望远镜和其他产品中使用的透镜的全三维形状进行了研究。预计这项工作将因此创造出新的和改进的产品和工艺，造福于英国工业和整个社会。

The Phase Focus Virtual Lens® (otherwise known as ptychography) is a new microscopymethod that eliminates the need for hardware focusing components, thereby removing thestraitjacket imposed by the costs, limitations and imperfections of conventional lenses.Instead of lenses, the Virtual Lens image reconstruction process is performed by asophisticated computer programme (algorithm.) Because it provides quantitative (numerical)information, and because it can extract the high information content often present intransparent specimens, the Virtual Lens has advantages in applications as diverse as live cellimaging and ophthalmic (contact) lens manufacture.The existing Virtual Lens algorithm relies on a two-dimensional mathematical approximationof the specimen. If the specimen’s structure is physically three-dimensional, or if it is thick,poor image quality can result. The purpose of this work, therefore, is to produce fulltomographic quantitative images of low-contrast specimens of complex structures, using anew three-dimensional Virtual Lens algorithm.If successful, it is expected that this new method will be able to generate high quality andaccurate information from such specimens without the use of stains, and from greater depthswithin the specimens than can be observed using conventional confocal microscopes.In addition, the technique is expected to be able to provide a new way to measure accurately,for manufacturing quality assurance purposes, the full three dimensional shape of lenses thatare used in CD and DVD players, mobile phones, microscopes, telescopes and other products.It is anticipated that this work will therefore create new and improved products and processesfor the benefit of UK industry and society at large.