Histology seminars

Yuankai Huo presents a computational workflow for turning whole-slide tissue images into quantitative, spatially resolved pathomics measurements. The session covers scalable feature extraction through spatial indexing, processing within computational regions, and matrix-based computation. It connects tissue morphology and spatial topology with biological and clinical outcomes, then considers generative artificial-intelligence methods for visualizing and interpreting correlated pathomics features. The emphasis is on scalable, explainable analysis for biomedical researchers.

High-quality glass lenses are commonplace in the design of optical instrumentation used across the biosciences. However, research-grade glass lenses are often costly, delicate and, depending on the prescription, can involve intricate and lengthy manufacturing - even more so in bioimaging applications. This seminar will outline 3D printing as a viable low-cost alternative for the manufacture of high-performance optical elements, where I will also discuss the creation of the world’s first fully 3D printed microscope and other implementations of 3D printed lenses. Our 3D printed lenses were generated using consumer-grade 3D printers and pose a 225x materials cost-saving compared to glass optics. Moreover, they can be produced in any lab or home environment and offer great potential for education and outreach. Following performance validation, our 3D printed optics were implemented in the production of a fully 3D printed microscope and demonstrated in histological imaging applications. We also applied low-cost fabrication methods to exotic lens geometries to enhance resolution and contrast across spatial scales and reveal new biological structures. Across these applications, our findings showed that 3D printed lenses are a viable substitute for commercial glass lenses, with the advantage of being relatively low-cost, accessible, and suitable for use in optical instruments. Combining 3D printed lenses with open-source 3D printed microscope chassis designs opens the doors for low-cost applications for rapid prototyping, low-resource field diagnostics, and the creation of cheap educational tools.

The aim of this half day virtual meeting is to consider what is currently achievable with existing techniques and to explore where advancements can be made in the short and medium term. Leading scientists in the field will highlight the questions currently being addressed using hard X-ray imaging techniques, volume electron microscopy and their combination with other imaging modalities, with a forward look to areas of opportunity becoming accessible as a result of the recent and upcoming synchrotron upgrades. We expect an exciting day filled with science focused talks and lively discussions on how the field will develop over the next few years.

The standard method for staining structures in the brain is to slice the brain into 2D sections. Each slice is treated using a technique such as in-situ hybridization to examine the spatial expression of a particular molecule at a given developmental timepoint. Depending on the brain structures being studied, slices can be made coronally, sagitally, or at any angle that is thought to be optimal for analysis. However, assimilating the information presented in the 2D slice images to gain quantitiative and informative 3D expression patterns is challenging. Even if expression levels are presented as voxels, to give 3D expression clouds, it can be difficult to compare expression across individuals and analysing such data requires significant expertise and imagination. In this talk, I will describe a new approach to examining histology slices, in which the user defines the brain structure of interest by drawing curves around it on each slice in a set and the depth of tissue from which to sample expression. The sampled 'curves' are then assembled into a 3D surface, which can then be transformed onto a common reference frame for comparative analysis. I will show how other neuroscientists can obtain and use the tool, which is called Stalefish, to analyse their own image data with no (or minimal) changes to their slice preparation workflow.

Von Economo stated that an "Ideal Cortical Map" would look very different to a parcellation. He suggested that an Ideal Cortical Map would involve the superimposition of many different cortical maps, with changes in each map shown at every single point. In line with this idea, I will discuss our recent research on identifying principal dimensions of cortical differentiation. In particular, I will highlight large-scale patterns of cytoarchitectural differentiation that can be observed using post mortem histology or in vivo microstructure-sensitive MRI. I aim to show how this approach provides a cohesive framework to understand cortical organisation across multiple biological scales. This allows us to formulate new ideas on the organisation and function of the brain regions (eg: mesiotemporal lobe), networks (eg: DMN) and the whole cortex.

Robust detection of biological structures such as neuronal dendrites in brightfield micrographs, tumor tissue in histological slides, or pathological brain regions in MRI scans is a fundamental task in bio-image analysis. Detection of those structures requests complex decision making which is often impossible with current image analysis software, and therefore typically executed by humans in a tedious and time-consuming manual procedure. Supervised pixel classification based on Deep Convolutional Neural Networks (DNNs) is currently emerging as the most promising technique to solve such complex region detection tasks. Here, a self-learning artificial neural network is trained with a small set of manually annotated images to eventually identify the trained structures from large image data sets in a fully automated way. While supervised pixel classification based on faster machine learning algorithms like Random Forests are nowadays part of the standard toolbox of bio-image analysts (e.g. Ilastik), the currently emerging tools based on deep learning are still rarely used. There is also not much experience in the community how much training data has to be collected, to obtain a reasonable prediction result with deep learning based approaches. Our software YAPiC (Yet Another Pixel Classifier) provides an easy-to-use Python- and command line interface and is purely designed for intuitive pixel classification of multidimensional images with DNNs. With the aim to integrate well in the current open source ecosystem, YAPiC utilizes the Ilastik user interface in combination with a high performance GPU server for model training and prediction. Numerous research groups at our institute have already successfully applied YAPiC for a variety of tasks. From our experience, a surprisingly low amount of sparse label data is needed to train a sufficiently working classifier for typical bioimaging applications. Not least because of this, YAPiC has become the "standard weapon” for our core facility to detect objects in hard-to-segement images. We would like to present some use cases like cell classification in high content screening, tissue detection in histological slides, quantification of neural outgrowth in phase contrast time series, or actin filament detection in transmission electron microscopy.

The inability to observe relevant biological processes in vivo significantly restricts human neurodevelopmental research. Advances in appropriate in vitro model systems, including patient-specific human brain organoids and human cortical spheroids (hCSs), offer a pragmatic solution to this issue. In particular, hCSs are an accessible method for generating homogenous organoids of dorsal telencephalic fate, which recapitulate key aspects of human corticogenesis, including the formation of neural rosettes—in vitro correlates of the neural tube. These neurogenic niches give rise to neural progenitors that subsequently differentiate into neurons. Studies differentiating induced pluripotent stem cells (hiPSCs) in 2D have linked atypical formation of neural rosettes with neurodevelopmental disorders such as autism spectrum conditions. Thus far, however, conventional methods of tissue preparation in this field limit the ability to image these structures in three-dimensions within intact hCS or other 3D preparations. To overcome this limitation, we have sought to optimise a methodological approach to process hCSs to maximise the utility of a novel Airy-beam light sheet microscope (ALSM) to acquire high resolution volumetric images of internal structures within hCS representative of early developmental time points.

Deep learning is an emerging computational approach inspired by the human brain’s neural connectivity that has transformed machine-based image analysis. By using histopathology as a model of an expert-level pattern recognition exercise, we explore the ability for humans to teach machines to learn and mimic image-recognition and decision making. Moreover, these models also allow exploration into the ability for computers to independently learn salient histological patterns and complex ontological relationships that parallel biological and expert knowledge without the need for explicit direction or supervision. Deciphering the overlap between human and unsupervised machine reasoning may aid in eliminating biases and improving automation and accountability for artificial intelligence-assisted vision tasks and decision-making. Aleksandar Ivanov Title:

Africa is blessed with a rich diversity and abundance in rodent and avian populations. This natural endowment on the continent portends research opportunities to study unique anatomical profiles and investigate animal models that may confer better neural architecture to study neurodegenerative diseases, adult neurogenesis, stroke and stem cell therapies. To this end, African researchers are beginning to pay closer attention to some of her indigenous rodents and birds in an attempt to develop spontaneous laboratory models for homegrown neuroscience-based research. For this presentation, I will be showing studies in our lab, involving cellular neuroanatomy of two rodents, the African giant rat (AGR) and Greater cane rat (GCR), Eidolon Bats (EB) and also the Striped Owl (SO). Using histological stains (Cresyl violet and Rapid Golgi) and immunohistochemical biomarkers (GFAP, NeuN, CNPase, Iba-1, Collagen 2, Doublecortin, Ki67, Calbindin, etc), and Electron Microscopy, morphology and functional organizations of neuronal and glial populations of the AGR , GCR, EB and SO brains have been described, with our work ongoing. In addition, the developmental profiles of the prenatal GCR brains have been chronicled across its entire gestational period. Brains of embryos/foetuses were harvested for gross morphological descriptions and then processed using immunofluorescence biomarkers to determine the pattern, onset, duration and peak of neurogenesis (Pax6, Tbr1, Tbr2, NF, HuCD, MAP2) and the onset and peak of glial cell expressions and myelination in the prenatal GCR. The outcome of these research efforts has shown unique neuroanatomical expressions and networks amongst Africa’s rich biodiversity. It is hopeful that continuous effort in this regard will provide sufficient basic research data on neural developments and cellular neuroanatomy with subsequent translational consequences.