Coplanar embedding of multiple 3D cell models in hydrogel towards high-throughput micro-histology.

Standardised and high-throughput methods have been developed for the production and experimental handling of some 3D in vitro models. However, adapted analytical tools are still missing for scientists and researchers to fully exploit the potential of complex cellular models in pre-clinical drug testing and precision medicine. Histology is the established, cost-effective and gold standard method for structural and functional tissue analysis. However, standard histological processes are challenging and costly to apply to 3D cell models, as their small size often leads to poor alignment of samples, which lowers analysis throughput. This body of work proposes a new approach: HistoBrick facilitates histological processing of spheroids and organoids by enabling gel embedding of 3D cell models with precise coplanar alignment, parallel to the sectioning plane, thus minimising the loss of sample material. HistoBrick's features are compatible with automation standards, potentially allowing automated sample transfer from a multi-well plate to the gel device. Moreover, HistoBrick's technology was validated by demonstrating the alignment of HepG2 cultured spheroids measuring 150-200 µm in diameter with a height precision of ± 80 µm. HistoBrick allows up to 96 samples to be studied across minimal sections, paving the way towards high-throughput micro-histology.

A Miniature Bio-Photonics Companion Diagnostics Platform for Reliable Cancer Treatment Monitoring in Blood Fluids.

In this paper, we present the development of a photonic biosensor device for cancer treatment monitoring as a complementary diagnostics tool. The proposed device combines multidisciplinary concepts from the photonic, nano-biochemical, micro-fluidic and reader/packaging platforms aiming to overcome limitations related to detection reliability, sensitivity, specificity, compactness and cost issues. The photonic sensor is based on an array of six asymmetric Mach Zender Interferometer (aMZI) waveguides on silicon nitride substrates and the sensing is performed by measuring the phase shift of the output signal, caused by the binding of the analyte on the functionalized aMZI surface. According to the morphological design of the waveguides, an improved sensitivity is achieved in comparison to the current technologies (<5000 nm/RIU). This platform is combined with a novel biofunctionalization methodology that involves material-selective surface chemistries and the high-resolution laser printing of biomaterials resulting in the development of an integrated photonics biosensor device that employs disposable microfluidics cartridges. The device is tested with cancer patient blood serum samples. The detection of periostin (POSTN) and transforming growth factor beta-induced protein (TGFBI), two circulating biomarkers overexpressed by cancer stem cells, is achieved in cancer patient serum with the use of the device.

Design and performance of an ultra-flexible two-photon microscope for in vivo research.

We present a cost-effective in vivo two-photon microscope with a highly flexible frontend for in vivo research. Our design ensures fast and reproducible access to the area of interest, including rotation of imaging plane, and maximizes space for auxiliary experimental equipment in the vicinity of the animal. Mechanical flexibility is achieved with large motorized linear stages that move the objective in the X, Y, and Z directions up to 130 mm. 360° rotation of the frontend (rotational freedom for one axis) is achieved with the combination of a motorized high precision bearing and gearing. Additionally, the modular design of the frontend, based on commercially available optomechanical parts, allows straightforward updates to future scanning technologies. The design exceeds the mobility of previous movable microscope designs while maintaining high optical performance.

Comment on 'A new method for fusion, denoising and enhancement of x-ray images retrieved from Talbot-Lau grating interferometry'.

In a recent paper (Scholkamm et al 2014 Phys. Med. Biol. 59 1425-40) we presented a new image denoising, fusion and enhancement framework for combining and optimal visualization of x-ray attenuation contrast, differential phase contrast and dark-field contrast images retrieved from x-ray Talbot-Lau grating interferometry. In this comment we give additional information and report about the application of our framework to breast cancer tissue which we presented in our paper as an example. The applied procedure is suitable for a qualitative comparison of different algorithms. For a quantitative juxtaposition original data would however be needed as an input.

A new method for fusion, denoising and enhancement of x-ray images retrieved from Talbot-Lau grating interferometry.

This paper introduces a new image denoising, fusion and enhancement framework for combining and optimal visualization of x-ray attenuation contrast (AC), differential phase contrast (DPC) and dark-field contrast (DFC) images retrieved from x-ray Talbot-Lau grating interferometry. The new image fusion framework comprises three steps: (i) denoising each input image (AC, DPC and DFC) through adaptive Wiener filtering, (ii) performing a two-step image fusion process based on the shift-invariant wavelet transform, i.e. first fusing the AC with the DPC image and then fusing the resulting image with the DFC image, and finally (iii) enhancing the fused image to obtain a final image using adaptive histogram equalization, adaptive sharpening and contrast optimization. Application examples are presented for two biological objects (a human tooth and a cherry) and the proposed method is compared to two recently published AC/DPC/DFC image processing techniques. In conclusion, the new framework for the processing of AC, DPC and DFC allows the most relevant features of all three images to be combined in one image while reducing the noise and enhancing adaptively the relevant image features. The newly developed framework may be used in technical and medical applications.

Reduction of phase artifacts in differential phase contrast computed tomography.

X-ray differential phase contrast computed tomography (DPC CT) with a Talbot-Lau interferometer setup allows visualizing the three-dimensional distribution of the refractive index by measuring the shifts of an interference pattern due to phase variations of the X-ray beam. Unfortunately, severe reconstruction artifacts appear in the presence of differential phase wrapping and clipping. In this paper, we propose to use the attenuation contrast, which is obtained from the same measurement, for correcting the DPC signal. Using the example of a DPC CT measurement with pronounced phase artifacts, we will discuss the efficiency of our phase artifact correction method.

Noise analysis of grating-based x-ray differential phase contrast imaging.

The sensitivity of x-ray radiographic images, meaning the minimal detectable change in the thickness or in the index of refraction of a sample, is directly related to the uncertainty of the measurement method. In the following work, we report on the recent development of quantitative descriptions for the stochastic error of grating-based differential phase contrast imaging (DPCi). Our model includes the noise transfer characteristics of the x-ray detector and the jitter of the phase steps. We find that the noise in DPCi depends strongly on the phase stepping visibility and the sample properties. The results are supported by experimental evidence acquired with our new instrument with a field of view of 50x70 mm(2). Our conclusions provide general guidelines to optimize grating interferometers for specific applications and problems.