Thin-section Techniques Research Articles

INFLUENCE AND PREDICTION OF SINTERED AGGREGATE SIZE DISTRIBUTION ON THE PERFORMANCE OF LIGHTWEIGHT ALKALI-ACTIVATED CONCRETE

The most effective method for mitigating the adverse environmental impacts of traditional concrete involves substituting cement and natural aggregate with waste and byproduct resources. Utilizing sintered lightweight aggregate (fly ash) in geopolymer concrete emerges as an efficient solution for managing and disposing of significant amounts of fly ash. The influence of sintered aggregate size distribution on the performance of alkali-activated concrete, focusing on compressive strength improvement. The study employs sintered fly ash aggregate (SFA) as coarse aggregate, aiming to optimize packing density through proper particle distribution. The highest compressive strength is achieved with a mix featuring 75% 4-8mm and 25% 8-12mm SFA. Regression-based strength models are developed, exhibiting good alignment with conventional concrete models. Thin section techniques reveal enhanced aggregate-matrix interaction due to the porous structure of SFA. The study emphasizes the potential of SFA in geopolymer concrete for sustainable construction. Lightweight geopolymer concrete, owing to its lower density, significantly reduces the overall structural load.

Method for macro-microscopic damage observation of rock hydraulic fracturing based on dic and thin section techniques

This study conducted systematic studies and in-depth analyses of micromechanics of hydraulically induced crack propagation. This study introduced a method for observing macromicroscopic damage in rock hydraulic fracturing based on DIC and thin-section techniques. We developed an advanced sealed visualization system equipped with high-speed cameras and macrolenses to facilitate precise observation of the hydraulic fracturing process. Additionally, Digital Image Correlation was used to accurately track and analyze the evolution of fractures. Our methodology clearly distinguished between intergranular and transgranular cracks using polarized light microscopy, thereby offering novel insights into grain-scale damage and strain localization within crystalline rocks. This research contributes to the understanding of the mechanisms underlying hydraulic fracturing at both the macroscopic and microscopic scales.

Microscopic enrichment and porosity-permeability reduction mechanism of residual oil in tight sandstone reservoirs: an insight from Chang 8 Member, Yanchang Formation, Ordos Basin, China

This study delves into the micro-occurrence states and enrichment mechanisms of residual oil, pivotal for advancing the production from tight sandstone reservoirs, particularly from the Chang 8 Member of the Upper Triassic Yanchang Formation, Ordos Basin. Through an analysis of 23 core samples, employing high-pressure mercury injection, field emission scanning electron microscopy, thin section, and X-ray diffraction techniques, distinct reservoir types were categorized. The utilization of environmental scanning electron microscope, multi-solvent continuous extraction, and an oil components separation system facilitated an intricate analysis of residual oil micro-occurrence states and their subsequent effects on porosity and permeability reduction across varying reservoir types. The findings accentuate the integral role of reservoir type in determining residual oil distribution within tight sandstone reservoirs. Favorable pore throat sorting and connectivity in specified reservoir types are identified as conducive to residual oil enrichment with a higher concentration of light components. In contrast, elevated carbonatite and clay content in other reservoir types leads to adsorption of heavy components, disrupting pore throat connectivity, and impeding crude oil filling. The varied interactions between oil and rock, oil–oil, and pore throat sealing significantly impact the distribution of oil components of residual oil, culminating in a notable reduction of porosity and permeability by 2.63% and 0.197 mD, with corresponding reduction rates of 27.19% and 46.69%, respectively. The insights derived from this study furnish a theoretical foundation for augmenting tight oil recovery and comprehending the enrichment mechanism of residual oil driven by the heterogeneity of tight sandstone reservoirs.

Deep Sandstone Reservoir Characteristics and Pore Structure Classification Based on Fractal Theory: A Case Study of the Bayingobi Formation in Guaizihu Sag, Yin'e Basin.

The paper focuses on deep oil and gas resources in the Bayingobi Formation of Guaizihu Sag in the Yin'e Basin. Previous studies overlooked the differences between the pores and throats, mainly focusing on pore analysis. This work aims to analyze the pore structure and petrophysical properties of the reservoir using various methods. The study utilized the constant velocity mercury intrusion method to quantify pores and throats separately. Scanning electron microscopy and casting thin section techniques characterize the pore and throat morphology. The analysis compares the pore structures in reservoirs with different petrophysical properties. Additionally, pore and throat types are classified based on fractal dimensions, and factors influencing their development are discussed. Results reveal feldspar lithic sandstone as the predominant rock type with a low compositional maturity. The sandstone reservoirs exhibit low porosity (10.23%) and ultralow permeability (0.99 mD). Primary reservoir pore spaces include intergranular pores, dissolution pores, and microfractures. Pore radius averages at 195.32 μm, while throat radius is 3.76 μm. Pore structures are categorized as micropore small-throat, small-pore small-throat, and large-pore coarse-throat types. The study area generally exhibits a high pore-to-throat ratio, impacting reservoir petrophysical properties significantly. Pore development is primarily influenced by early diagenesis, organic acid dissolution, and hydrocarbon filling. Weak compaction and cementation transformations provide a material and spatial basis for the subsequent dissolution. The presence of thick organic-rich mudstone above and below the reservoir contributes to organic acid dissolution and hydrocarbon filling.

The term “classic invasive lobular carcinoma” of the breast defines breast malignancies of vastly different nature

PurposeTo describe in detail the special features of a previously unappreciated “classic invasive lobular carcinoma” which is confined to the terminal ductal lobular units (TDLUs) and differs considerably from the extensive classic invasive lobular carcinoma, and to suggest specific terminology. MethodAll invasive breast cancer cases without associated microcalcifications diagnosed in our Institution with the histopathologic diagnosis of classic invasive lobular carcinoma during the years 1996–2019 (n = 560) formed the basis of this study. The cases were prospectively classified according to their imaging biomarkers (mammographic features) and followed up to Dec 31, 2021, to determine long-term patient outcome. An additional 2600 invasive breast cancer cases (diagnosed other than invasive lobular carcinoma) without associated microcalcifications served as a reference group. Detailed histopathologic analysis used large format (10x8 cm) thin section technique and staining methods including hematoxylin-eosin (H&E), E-cadherin, cytokeratin CK 5/6, a transmembrane glycoprotein (CD44) and anti-actin or anti-smooth muscle myosin heavy chain. ResultsThe imaging biomarkers differentiated two separate disease subgroups, having the same histopathologic diagnosis, classic invasive lobular carcinoma. One of these has the imaging biomarker of extensive architectural distortion with no central tumour mass, occupies the extralobular mesenchyme and has a long-term survival of 56%. The other subgroup forms stellate or circular non-calcified tumour masses usually smaller than 20 mm, which appear to arise in the intralobular mesenchyme, and has a significantly better long-term survival of 84%. ConclusionsThere is a striking difference between the subgross histopathology and the mammographic appearance (imaging biomarkers) of two breast malignancies having the same histopathologic diagnosis, “classic invasive lobular carcinoma”. The large difference in the long-term outcome of these two tumour types is even more striking. Using the same specific term, “classic invasive lobular carcinoma”, to describe these two separate entities can adversely affect management decisions.

First discovery of parasite eggs in a vertebrate coprolite of the Late Triassic in Thailand.

A paleoparasitological investigation of a vertebrate coprolite from the Huai Hin Lat Formation (Upper Triassic) was carried out. Five morphotypes of potential parasite eggs or sporocysts were identified in the coprolite by microscopic analysis using thin section technique. The rounded or oval shape and thick shell of one of the five morphotypes suggests that it belongs to nematode of the order Ascaridida. Systematic assignment of other morphotypes cannot be done in detail but suggests that the host was parasitized by different species of parasites. This is the first record of parasites in terrestrial vertebrate hosts from the Late Triassic in Asia and it provides new information on parasite-host interactions during the Mesozoic era.

Podocarpus (Podocarpaceae) wood from miocene rocks in Panotla, Tlaxcala, Mexico

Permineralized fossil woods were discovered in a Miocene sedimentary lake sequence in Panotla, Tlaxcala. The objective of this work is to present the description and identification of permineralized woods found in Miocene strata of Tlaxcala and discuss their biogeographical importance. The fossils were prepared using the thin section technique and identified using wood anatomy. The woods were identified as belonging to the genus Podocarpus, with a new species, Podocarpus mexicanoxylon sp. nov Castañeda-Posadas. This record is of great importance because: a) Its presence demonstrates the presence of the ecotone of the tropical montane cloud forest and b) This report supports the presence of this genus at low latitudes in North America (Mexico), where it was unknown until now.

An Overview of Laboratory Diagnosis of Central Nervous System Viral Infections

Central nervous system (CNS) infection is a serious illness that can lead to death. CNS infections include meningitis, encephalitis, brain abscesses and myelitis. These diseases are caused by causative agents like bacteria, fungi, parasites, and protozoa, but most commonly by viral infections. To combat this issue, accurate diagnosis of etiological agents at an early stage is crucial for appropriate treatment, control of the disease and prevent from becoming life-threatening to the patients. This review paper summarises the main laboratory diagnostic methods for CNS infections caused by viruses ranging from conventional to molecular methods. Conventional isolation methods are considered the ‘gold standard’ as they provide accurate evidence, but require highly skilled personnel, are time-consuming, critical in cell type selection and are useless for non-cultivable viruses. Electron microscopy allows recognition of viral morphology and ultrastructural details as the principle of virus identification through negative staining or thin section technique (suitable for tissue or cell specimens). However, it offers low sensitivity and requires at least 106 virions per millilitre or milligram in the specimen to be detectable by microscopy. Immunological-based methods have been extensively applied for viral diagnosis by detecting the antiviral antibodies or viral antigens in clinical samples. While these methods provided high sensitivity and specificity, the incubation and window period of an infection may give false-negative results. Lastly, molecular detections have many advantages such as high sensitivity, specificity, rapid, require a small amount of sample, simultaneous detection of multiple different viruses, and produce both qualitative and quantitative results.

Optimization and Prediction of Concrete with Recycled Coarse Aggregate and Bone China Fine Aggregate Using Response Surface Methodology

Construction recycled material is crucial for protecting natural resources and promoting sustainable human development in a rapidly industrializing world. Many administrations worldwide accepted that it is beneficial to use demolition waste in the concrete building industry to reduce manufacturing costs and minimize the use of virgin aggregates. However, control measures should be done as their mechanical properties are poorer than traditional aggregates. To overcome this problem, pozzolanic materials like bone chine can be incorporated, providing extra CSH gel, which improves mechanical strength. Therefore, this research is aimed at producing eco‐friendly concrete, which can be used for medium‐grade strength, using recycled construction waste (RCA) as coarse and bone china fine aggregate (BCA) as fine aggregate. Workability, density, compressive, split tensile, and flexural strength are used to compare the fresh and hardened properties of the concrete. Experimental and statistical research is employed in the current study to evaluate the impact of RCA and BCA on the performance of concrete. To simulate all measurable responses, including workability, density, compressive, flexural, and split strength, RSM (response surface methodology) was utilized. The CCD (Central Composite Design) approach in RSM was used to create and analyze mixes in an experiment. Based on the experiment’s results, mathematical models were designed and assessed using the analysis of variance test (ANOVA). The analysis of variance results demonstrated the statistical significance of each constructed model. Three‐dimensional response surface plots created using established regression models were used to investigate the interaction between the respective variables and to optimize the mixing ratio. The results indicate that the optimum utilization of RCA is up to 40% and BCA up to 60% as coarse and fine aggregate replacement in concrete, respectively, which not only helps to reduce costs but also offers sustainability. Finally, it was concluded that the generated models might be employed by obtaining the maximum tested features of concrete to assure a quick mix design approach. To conduct the microstructure study, thin section techniques were used to observe a strong aggregate‐matrix interaction.

Effects of Diagenesis on the Reservoir Quality in the Upper Sands of Lower Goru Formation, Badin Block, Lower Indus Basin, Pakistan

The core samples of B member of upper sand reservoir rocks of the lower Goru Formation from three wellsof the Badin block were studied using thin section, XRD, and SEM techniques to investigate the diagenetic trends andtheir effect on reservoir quality. Microscopic study indicates that the B sand unit is mature with quartz as thepredominant mineral constituent with variable amount of feldspar and lithic minerals. The QFL plot indicates that mostof the samples are plotted in the field of quartz arenite, sub-litharenite and sub-arkose respectively. A few samplesbelong to the category of litharenite and feldspathic litharenite. The feldspars were partially to completely altered tokaolinite and other clay minerals. Coarse-crystalline or micro-crystalline calcite is the predominant cementing material.Bulk rock XRD analysis also confirms that the main mineral constituents of there samples are quartz and calcite invariable proportions. The undulose extinction and fracturing of quartz grains indicate that the area remained understress. Moreover, such fracturing is post-depositional and therefore is the product of late diagenesis. Scanning ElectronMicroscopic (SEM) images at 50 micrometer (μm) size show irregular type of fracturing within the quartz grains. Thislate stage fracturing of quartz has also generated various types of channels which may serve as secondary porosity. Thequartz overgrowth was observed in some samples due to late stage diagenesis. The micro-crystalline cement in the formof calcite is mostly present within the pores in fractured quartz. The results indicate that the diagenesis played asignificant role in improving the reservoir characteristics of B sand by increasing the porosity due to quartz fracturing,feldspar dissolution, alteration and quartz corrosion. Hence, better understanding of reservoir heterogeneities inporosity through diagenetic studies can be helpful in evaluating potential reservoir horizons for hydrocarbonaccumulation on spatial and temporal scales.

Platelet Electron Microscopy: Utilizing LEAN Methodology to Optimize Laboratory Workflow.

Platelet electron microscopy (PEM) is the gold standard methodology for diagnosing storage pool disorder, defined as a paucity of delta granules, alpha granules, or both. PEM literature is limited with few published resources and without well-developed interlaboratory standardization for the preparation and examination of platelet samples. Whole mount (WM) dense body (DB) counts for 300 pediatric cases were reviewed to determine whether counting fewer platelets could yield the same results. For 6 cases, DB average was determined on the day of WM preparation and on 2 consecutive days. Both WM and thin section (TS) preparations were examined for all cases. Employing LEAN methodology, an algorithm was developed to markedly decrease the number of platelets counted and still ensure accurate results. WMs decay with time; a statistically significant difference in DB counts was noted between day 0 and day 1 (p < .1). Twelve of 300 cases required both WM and TS preparations for a complete diagnosis. It is possible to maintain accuracy and decrease 100 platelet DB counts by >75%. WMs must be counted on the day they are prepared to avoid false paucity of DB secondary to sample decay. An accurate evaluation of platelet morphology requires both the WM and TS techniques.

Flow Unit Model of Channel Sand Body and Its Effect on Remnant Oil Distribution: A Case Study of PI Formation in the Eastern Transition Zone of Daqing Oilfield

To analyze the effect of various flow units in a channel sand body on remnant oil, we established a connection between various flow unit types and the remnant oil distribution. Using stratigraphic correlation and the characterization of sedimentary microfacies, we describe a single sand body, point bar, and narrow channel located at the injection-production well pattern of well B2-60-FB271 in the Eastern transition zone of the Daqing Placanticline. Architecture models of the point bar and narrow channel are also established using a series of parameters from different measurement methods. Four types of flow units (strong-current limiting, medium-current limiting, weak-current limiting, and none-current limiting) in the point bar sand body were identified, whereas one type, unshielded unit, was identified in the narrow channel. Geological parameters, such as porosity, permeability, and pore-throat radius (r50), were optimized to quantitatively characterize these various flow units. Samples were obtained from well B2-60-FB271 and analyzed by the freeze-fluorescence thin section technique. According to the displacement degree, the microscopic remnant oil was divided into three types: (1) free-state remnant oil, (2) semi-free-state remnant oil, and (3) bound-state remnant oil. We found that the strong-current limiting flow unit in the point bar is the enrichment area of free-state microscopic remnant oil and that the medium-current limiting and weak-current limiting flow units also have relatively high free microscopic remnant oil. These constitute the remaining oil enrichment areas in the study area.

A New Coryphoid Palm from the Maastrichtian-Danian Sediments of Madhya Pradesh and its Palaeoenvironmental Implications

Abstract A new petrified palm stem Palmoxylon dindoriensis Khan, Roy et Bera, sp. nov. from the latest Maastrichtian (late Cretaceous)-earliest Danian (early Paleocene) sediments of the Deccan Intertrappean beds of Madhya Pradesh, central India is reported. Sections from different parts of the recovered stem were prepared by using standard thin section techniques and studied under transmitted light compound and scanning electron microscopes. The significant anatomical characters of the present fossil stem are the presence of well-preserved fibrovascular bundles (fvbs) with reniform type of dorsal fibrous sclerenchymatous part (dcap), two metaxylem vessel elements in each fvb, well-developed two-layered tabular parenchyma around dcap of fvb, fibrous bundles throughout the CZ, highly compact ground parenchyma tissue and the absence of centrifugal differentiation of fibrous part of fvb. The aforesaid anatomical attributes reveal its close resemblance with the extant taxa of subfamily Coryphoideae of the family Arecaceae having Cocos-type general stem organization. The core distribution of modern palms and the presence of highly compact ground parenchyma of the present fossil suggest that they thrived under a tropical, terrestrial warm, humid environment during the time of deposition (latest Maastrichtian-earliest Danian).

Technique for preparation thin sections of structural soil aggregates of certain size fractions

Morphological properties of poil aggregate aer developed in the process of pedogenesis and reflect the complex of structure formation processes. The peds' size, shape, and internal structure are peculiar to each corresponding soil type and genetic horizon. The technique proposed allows preparing thin sections on every sizes of structural units and, in contrast to known methods, it provides not only a ped immobilization in the mass of fixing material, but also pore space saturation with natural resin. It is also possible to prepare the sections with marking of large aggregates orientation in space; it requires application of gypsum marks on the aggregate surface in the process of their sampling from the soil profile. The technique modified consists of following stages: selection of aggregates with required fractions; preparation of sections with separate aggregates larger than 5 mm; preparation of sections with aggregations smaller than 5 mm. In preparation of aggregates larger than 5 mm, a technique similar to that of preparing large soil block samples is used because it makes possible to process each soil aggregate separately. Another approach is required for processing of structural units up to 5 mm in size because it is impossible to prepare a section of individual issue with this size. The problem is solved when the units are saturated and processed not separately, but as a whole block sample which include a mass with peds of separate fraction. For this purpose, it is required to prepare cups comprised of walls from thick paper and a gypsum bottom. For preparation the caps use a gypsum in the form of fine white powder rapidly solidified in addition of water in a certain proportion and forms a firm bottom of the paper сup. Cut out a square of 4×4 cm in size and a strip from thick paper and glue them together to gain a tube of 5 cm in height and 1.5–2 cm in diameter (i.e. 10×5 cm). Apply on the paper squares a prepared gypsum mass sufficient to retention of the paper tube until gypsum hardening. Fill a fraction of aggregates into the prepared cup in one-third of its height and cover it with prepared fixing substance with solvent. Saturation is carried out in accordance with the standard technique of section preparing using natural resins (Gagarina, 2004). After complete saturation, paper cups with a mass of aggregates in them turn into monoliths that should be processed in the same way as large soil samples. As a result of all operations, significant number of structural soil aggregates with same sizes may be found within the section plane.

Evidence of simultaneous occurrence of tylosis formation and fungal interaction in a late Cenozoic angiosperm from the eastern Himalaya

Information on whether tyloses in fossil angiospermic plants form specifically in response to infestation with pathogenic fungi has been lacking so far, and thus the evolutionary history of tylosis formation as a physical restraint strategy against the advancing fungi remains unresolved. Our study addresses this knowledge lacuna. Carbonized woods were recovered from the upper part of the Siwalik succession (Kimin Formation: late Pliocene to early Pleistocene) of Arunachal Pradesh, eastern Himalaya. Samples from different portions of the wood remains were prepared by using standard thin sectioning techniques and studied under transmitted light compound and scanning electron microscopes. The Plio-Pleistocene wood remains are attributed to Gmelina arborea Roxb. (Lamiaceae) as Gmelina siwalika Khan, Bera M et Bera S, sp. nov. Numerous well-preserved tyloses were found mainly in the central heartwood region of the fossil wood. Rare fungal remains indicative of early-stage colonization are characterized by septate fungal hyphae extending through the vascular ray system within the secondary xylem, and fungal spores. As tyloses are present in large numbers and formed before the spread of fungal remains, they served as an effective physical restraint to the spread of the fungi. We conclude that tylosis formation contributed to an antagonistic relationship between pathogenic fungi and Gmelina plant in Neogene paleoecosystems of the eastern Himalaya. This report also demonstrates that the development of this histological protrusion has been a significant process in woody plants since Cenozoic.

Mineralogy and spectroscopy of Owen Group sandstones, Australia: implications for the provenance, diagenesis, and origin of coloration

Mineralogical studies were conducted on sandstones of Late Cambrian-Early Ordovician Owen Group, southern West Coast Range, Tasmania, Australia. Several samples were analysed using conventional thin section and point counting techniques, while others were further analysed under a Scanning Electron Microscope with back scattering and Energy Dispersive X-ray Spectroscopy capabilities. The result of petrographic analyses includes provenance, possible sources and controls over coloration in sandstone samples. The sandstones are mainly comprised of quartz, mica, lithic grains (volcanic, metamorphic and sedimentary), clays, zircon, tourmaline, leucoxene, rutile, and chert grains. These were derived from a Proterozoic Tyennan metamorphic and a Middle Cambrian Mount Read Volcanic provenance. Following deposition, sediments underwent severe diagenesis including quartz overgrowth, iron coatings, haematite, chlorite and sericite alterations, linings and cements. Multi-phase haematite-related diagenesis is present and late stage hydrothermal fluid alterations assemblages have severely altered and colored the sediments.

Under pressure? Epicormic shoots and traumatic growth zones in high-latitude Triassic trees from East Antarctica.

Investigating the biology of trees that were growing at high latitudes during warmer geological periods is key to understanding the functioning of both past and future forest ecosystems. The aim of this study is to report the first co-occurrence of epicormic shoots and traumatic growth zones in fossil trees from the Triassic of Antarctica and to discuss their biological and environmental implications. Permineralized woods bearing scars of epicormic shoots were collected from the Triassic Fremouw Formation in Gordon Valley, Central Transantarctic Mountains, Antarctica in 2010. Samples from different portions of three specimens were prepared using standard thin section and hydrofluoric (HF) acid peel techniques, and anatomical details were studied in transmitted light. The fossil woods represent the outer part of trunks, with at least 40 growth rings that are 0.2-4.8 mm in width. Anatomical comparisons suggest that they represent a new tree taxon for the Triassic of Antarctica. Numerous small epicormic shoots can be seen crossing the wood almost horizontally and are locally branched. Each specimen also contains several occurrences of traumatic growth zones located in the early wood, in the cells produced either at the very start of the growing season or slightly later. This is the first report of epicormic shoots and traumatic growth zones in the wood of a Triassic tree from Antarctica. Their co-occurrence indicates that these trees from Gordon Valley were subjected to environmental stresses not seen in Triassic trees previously described from this region. This suggests that they had a different biology and/or were growing in a different habitat, which offers a new glimpse into the diversity of high-latitude trees in the Triassic greenhouse climate.

Discovery of Miocene Wood from Manchar Formation, Lagerstroemioxylon thanobolensis Sp. Nov. of Family Lythraceae , Thanobola Khan District Jamshoro, Sindh, Pakistan

The paper presents taxonomic identifications by the paleoxylotomical studies of petrified fossil wood collected from Manchar Formation of Miocene age exposed in Thanobola khan Sindh Pakistan. Sections were prepared by standard ground thin section techniques. Anatomical characters revealed from the microscopic study were used to compare with modern wood and already reported fossil woods. The sample was found comparable with family Lytheraceae and with species in respect of character of Vessels, arrangements of wood Parenchyma and Xylem rays. Therefore it is given as a name Lagerstroemioxylon thanobolensis sp.nov., represent the locality of Thano Bola Khan

Abstract 5915: Three-dimensional, 3-D, multiplex imaging of biomarkers in tumor tissue

Abstract Background: Obtaining high-resolution information from solid tumors, while maintaining the spatial perspective needed to understand the heterogeneous tumor microenvironment, represents a key challenge for preclinical and clinical cancer applications. Methods: We utilized a novel method CLARITY that has been shown to transform intact rodent tissues into an optically transparent and permeable hybridized-hydrogel form that can undergo immunostaining followed by 3-D imaging of nucleic acids and proteins markers, without the need to separately analyze hundreds of conventional FFPE thin sections. In this study, human breast and lung cancer excision biopsy samples were obtained commercially. Murine orthotopic MCF-7 xenograft and patient-derived-xenograft (PDX) breast tumors were explanted and prepared as fresh samples. The intact tissues were incubated in a mixture of 4% paraformaldehyde/4% acrylamide/0.05% bis-acrylamide for 48 hours, and polymerized in the presence of a thermal initiator, VA-044, for 3 hours at 37°C to form an intact hydrogel/tissue matrix. The tissues were then lipid-cleared in a solution of 0.2M borate buffer containing 8% SDS, pH 8.5 at 45-55°C from 5-35 days, depending on sample size. The cleared tissues were then stained with two multiplex antibody panels that were selected to highlight the microenvironment of the tissue including, immune cells (CD-3 or CD-8), vasculature (Lectin-Texas Red or CD-31), and tumor (cytokeratin [8+18] or PD-L1, directly conjugated to Alexa Fluor® 647). For non-directly conjugated antibodies, the tissues were then subjected to a secondary goat anti-mouse or goat anti-rabbit antibody conjugated to either Alexa Fluor® 568 or 700 followed by a nuclear counterstain, Sytox Blue. Samples were then placed in an appropriate refractive index solution for imaging on a laser scanning confocal microscope. Results: The tissues remained intact throughout the procedure and the cellular morphology of the tissue was well preserved. The individual components of the microenvironment could be identified, demonstrating that the tissues could be successfully interrogated with a multiplex of indirect and directly conjugated commercially available antibodies. These preliminary studies, provide evidence for the use of this novel technology for detecting the heterogeneity of biomarker expression within the tumor microenvironment. Most notably, the analysis of a lymph node from a patient with metastatic breast cancer revealed variation across the tissue which may have implications for the detection of micro-metastasis in patients with early stage disease when using conventional thin section histological methods. Conclusion: These results implicate CLARITY as a powerful next generation tissue processing technology for profiling the intact tumor microenvironment, eliminating the need to recapitulate this spatial and quantitative information with standard thin section techniques. Citation Format: Sharla L. White, Sarah McCurdy, Laurie J. Goodman. Three-dimensional, 3-D, multiplex imaging of biomarkers in tumor tissue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5915. doi:10.1158/1538-7445.AM2017-5915

Synchrotron computer tomographic (CT) scans complement traditional techniques in understanding the internal anatomy of permineralised Fontainocarpa (Crotonoideae, Euphorbiaceae) fruits from the Oligocene of eastern Australia

The internal morphology and anatomy of silicified fruits of Fontainocarpa were studied using traditional thin sectioning techniques, SEM and synchrotron computed tomographic (CT) imaging and animations, to enable comparative analyses with extant, indehiscent-fruited genera in the Euphorbiaceae including Fontainea, Aleurites and Hylandia from Australia, and other non-Australian crotonoid genera. Thin sections and sectioning show that the fruits of Fontainocarpa are indehiscent, multicarpellate and usually 3- to 5-loculate, with axial placentation, a single ovule per carpel and the ovules are anatropous and have antitropous curvature. A ventral vascular trace that supplies each ovule is embedded in the bitegmic seed coat. The internal anatomy is therefore consistent with the Euphorbiaceae. Additional characters, including indehiscent fruits, distinctive vascular channels (foramina) that penetrate through the fruit wall into the locule, and thin membranous seed coats are restricted to very few genera in the Euphorbiaceae, but occur together in extant Fontainea. The seed coat in extant Fontainea and fossil Fontainocarpa seeds is membranous, and appears to lack the palisadal exotegmen of most genera in the Euphorbiaceae. Fontainocarpa fruits were compared with those of extant Fontainea and the fossil has a combination of features unlike those of extant taxa. It shares with Fontainea picrosperma in having endocarps with convex intersutural surfaces lacking ornamentation and a similar number of locules and with Fontaineavenosa in having conspicuous foramina. This study therefore supports a close relationship between Fontainea and Fontainocarpa and is further evidence of the Crotonoideae in the fossil record in Australia, and is one of the few records of this subfamily worldwide. This study is one of the few, to date, using synchrotron CT imaging to reveal the internal morphology of silicified fruits and to utilize animations to examine the structure of these fruits.