Abstract. In cancer treatment, it is highly desirable to classify single cancer cells in real time. The standard method is polymerase chain reaction requiring a substantial amount of resources and time. Here, we present an innovative approach for rapidly classifying different cell types: we measure the diffraction pattern of a single cell illuminated with coherent extreme ultraviolet (XUV) laser-generated radiation. These patterns allow distinguishing different breast cancer cell types in a subsequent step. Moreover, the morphology of the object can be retrieved from the diffraction pattern with submicron resolution. In a proof-of-principle experiment, we prepared single MCF7 and SKBR3 breast cancer cells on gold-coated silica slides. The output of a laser-driven XUV light source is focused onto a single unstained and unlabeled cancer cell. With the resulting diffraction pattern, we could clearly identify the different cell types. With an improved setup, it will not only be feasible to classify circulating tumor cells with a high throughput, but also to identify smaller objects such as bacteria or even viruses.

BackgroundDiagnosis of intestinal metaplasia and dysplasia via conventional endoscopy is characterized by low interobserver agreement and poor correlation with histopathologic findings. Chromoendoscopy significantly enhances the visibility of mucosa irregularities, like metaplasia and dysplasia mucosa. Magnetically guided capsule endoscopy (MGCE) offers an alternative technology for upper GI examination. We expect the difficulties of diagnosis of neoplasm in conventional endoscopy to transfer to MGCE. Thus, we aim to chart a path for the application of chromoendoscopy on MGCE via an ex-vivo animal study.MethodsWe propose a modified preparation protocol which adds a staining step to the existing MGCE preparation protocol. An optimal staining concentration is quantitatively determined for different stain types and pathologies. To that end 190 pig stomach tissue samples with and without lesion imitations were stained with different dye concentrations. Quantitative visual criteria are introduced to measure the quality of the staining with respect to mucosa and lesion visibility. Thusly determined optimal concentrations are tested in an ex-vivo pig stomach experiment under magnetic guidance of an endoscopic capsule with the modified protocol.ResultsWe found that the proposed protocol modification does not impact the visibility in the stomach or steerability of the endoscopy capsule. An average optimal staining concentration for the proposed protocol was found at 0.4% for Methylene blue and Indigo carmine. The lesion visibility is improved using the previously obtained optimal dye concentration.ConclusionsWe conclude that chromoendoscopy may be applied in MGCE and improves mucosa and lesion visibility. Systematic evaluation provides important information on appropriate staining concentration. However, further animal and human in-vivo studies are necessary.

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In cancer treatment it is highly desirable to identify and /or classify individual cancer cells in real time. Nowadays, the standard method is PCR which is costly and time-consuming. Here we present a different approach to rapidly classify cell types: we measure the pattern of coherently diffracted extreme ultraviolet radiation (XUV radiation at 38nm wavelength), allowing to distinguish different single breast cancer cell types. The output of our laser driven XUV light source is focused onto a single unstained and unlabeled cancer cell, and the resulting diffraction pattern is measured in reflection geometry. As we will further show, the outer shape of the object can be retrieved from the diffraction pattern with sub-micron resolution. For classification it is often not necessary to retrieve the image, it is only necessary to compare the diffraction patterns which can be regarded as a spatial fingerprint of the specimen. For a proof-of-principle experiment MCF7 and SKBR3 breast cancer cells were pipetted on gold-coated silica slides. From illuminating each single cell and measuring a diffraction pattern we could distinguish between them. Owing to the short bursts of coherent soft x-ray light, one could also image temporal changes of the specimen, i.e. studying changes upon drug application once the desired specimen is found by the classification method. Using a more powerful laser, even classifying circulating tumor cells (CTC) at a high throughput seems possible. This lab-sized equipment will allow fast classification of any kind of cells, bacteria or even viruses in the near future.