High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials

Geyer, Stefan H.; Maurer‐Gesek, Barbara; Reissig, Lukas F.; Weninger, Wolfgang J.

doi:10.3791/56071

Cited by 29 publications

(45 citation statements)

References 47 publications

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“…; Mohun & Weninger, ; Geyer et al. ). The material was removed from the storing solution and rinsed under running tap water for 12 h. Then it was dehydrated in a series of ethanols (50%, 70%, 80%, 90%, 96%).…”

Section: Methodsmentioning

confidence: 99%

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The dermal arteries in the cutaneous angiosome of the descending genicular artery

et al. 2018

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Studies examining thick skin of the thumb pad have challenged the existence of an arterial plexus in the papillary dermis. Instead of a plexus, discrete arterial units, interconnected by arterio-arterial anastomoses, were identified. We hypothesise that the dermal arteries of thin skin are arranged likewise and that there are fewer arterio-arterial anastomoses in the centre of an angiosome than in zones where neighbouring angiosomes overlap. To test these hypotheses, we examined the dermal arteries in the centre of the cutaneous angiosome of the descending genicular artery (DGA) and its zone of overlap with neighbouring angiosomes. Using traditional perfusion techniques, the cutaneous angiosomes of the DGA and the popliteal artery were identified in 11 fresh frozen human lower limbs. Biopsies were harvested from the centre of the cutaneous DGA angiosome and from the zone where neighbouring vascular territories overlapped. Employing high-resolution episcopic microscopy (HREM), digital volume data were generated and the dermal arteries were three-dimensionally reconstructed and examined. In all examined skin areas, the dermal arteries showed tree-like ramifications. The branches of the dermal arteries were connected on average by 1.73 ± 1.01 arterio-arterial anastomoses in the centre of the DGA angiosome and by 3.27 ± 1.27 in the zone where angiosomes overlapped. We demonstrate that discrete but overlapping dermal arterial units with a mean dimension of 1.62 ± 1.34 and 1.80 ± 1.56 mm , respectively, supply oxygen and nutrients to the superficial dermis and epidermis of the thin skin of the medial femur. This forms the basis for diagnosing and researching skin pathologies.

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Section: Methodsmentioning

confidence: 99%

“…; Mohun & Weninger, ,c; Geyer et al. ). Each volume dataset comprised 2000–2300 inherently aligned digital images.…”

Section: Methodsmentioning

confidence: 99%

The dermal arteries in the cutaneous angiosome of the descending genicular artery

et al. 2018

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“…Data generation was performed as described by Geyer et al (). Briefly, blocks were mounted on the robotic microtome HREM unit (Indigo Scientific) equipped with MVX10 Olympus Microscope (Olympus) and ProgRes MFscan digital camera (Jenoptic).…”

Section: Methodsmentioning

confidence: 99%

“…However, these techniques are laborious, prone to alignment errors, and result in 3D models with lower resolution quality compared with the 2D sections, partially due to the effect of tissue sectioning and staining (Mohun & Weninger, ). To overcome these obstacles, a new technique for whole‐mount 3D imaging termed high‐resolution episcopic microscopy (HREM) was developed (Geyer, Maurer‐Gesek, Reissig, & Weninger, ; Geyer et al, ; Mohun & Weninger, ; Weninger et al, , ). HREM is a microscope–microtome‐based imaging system, which generates hundreds to thousands of perfectly aligned thin‐section images of an embedded tissue.…”

Section: Introductionmentioning

confidence: 99%

High‐resolution episcopic microscopy enables three‐dimensional visualization of plant morphology and development

et al. 2019

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The study of plant anatomy, which can be traced back to the seventeenth century, advanced hand in hand with light microscopy technology and relies on traditional histologic techniques, which are based on serial two‐dimensional (2D) sections. However, these valuable techniques lack spatial arrangement of the tissue and hence provide only partial information. A new technique of whole‐mount three‐dimensional (3D) imaging termed high‐resolution episcopic microscopy (HREM) can overcome this obstacle and generate a 3D model of the specimen at a near‐histological resolution. Here, we describe the application of HREM technique in plants by analyzing two plant developmental processes in woody plants: oil secretory cavity development in citrus fruit and adventitious root formation in persimmon rootstock cuttings. HREM 3D models of citrus fruit peel showed that oil cavities were initiated schizogenously during the early stages of fruitlet development. Citrus secretory cavity formation, shape, volume, and distribution were analyzed, and new insights are presented. HREM 3D model comparison of persimmon rootstock clones, which differ in their rooting ability, revealed that difficult‐to‐root clones failed to develop adventitious roots due to their inability to initiate root primordia.

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“…After polymerisation at room temperature, the resin blocks were baked at 90 °C for 1–2 days and used for HREM data generation (Mohun & Weninger, ; Geyer et al. ). Resulting digital datasets consisted of approximately 3000–4000 precisely aligned single images and had isotropic voxel sizes with edge lengths between 2.5 and 3.5 μm.…”

Section: Methodsmentioning

confidence: 99%

Morphology, topology and dimensions of the heart and arteries of genetically normal and mutant mouse embryos at stages S21–S23

et al. 2017

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Accurate identification of abnormalities in the mouse embryo depends not only on comparisons with appropriate, developmental stage‐matched controls, but also on an appreciation of the range of anatomical variation that can be expected during normal development. Here we present a morphological, topological and metric analysis of the heart and arteries of mouse embryos harvested on embryonic day (E)14.5, based on digital volume data of whole embryos analysed by high‐resolution episcopic microscopy (HREM). By comparing data from 206 genetically normal embryos, we have analysed the range and frequency of normal anatomical variations in the heart and major arteries across Theiler stages S21–S23. Using this, we have identified abnormalities in these structures among 298 embryos from mutant mouse lines carrying embryonic lethal gene mutations produced for the Deciphering the Mechanisms of Developmental Disorders (DMDD) programme. We present examples of both commonly occurring abnormal phenotypes and novel pathologies that most likely alter haemodynamics in these genetically altered mouse embryos. Our findings offer a reference baseline for identifying accurately abnormalities of the heart and arteries in embryos that have largely completed organogenesis.

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High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials

Cited by 29 publications

References 47 publications

The dermal arteries in the cutaneous angiosome of the descending genicular artery

The dermal arteries in the cutaneous angiosome of the descending genicular artery

High‐resolution episcopic microscopy enables three‐dimensional visualization of plant morphology and development

Morphology, topology and dimensions of the heart and arteries of genetically normal and mutant mouse embryos at stages S21–S23

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