Translucent Tissue Research Articles

Ayurvedic management of Nasa Arsha w.s.r. to Nasal Polyp: A Case Report

Nasa Arsha is one among the Nasagata Roga explained in Ayurvedic classics. When the Prakupit Dosha gets localised in the Ghran Pradesh, the person gets the symptoms like Pratishyaya (cold), Atim Matra Kshavathu (excessive sneezing), Kruchchhra Uchchhawas (difficulty in breathing), Putinasya (fetor nasalis), Shirah Shul (headache), Anunasik Vakya Dukha (difficulty in nasal letters). Nasal polyp is oedematous grape-like protrusions most often originating in the upper part of the nose around the osteomeatal complex on the lateral wall. The surface epithelium tends to be smooth and consists of pale translucent tissue which distinguishes them from the more vascular mucosa of the nasal cavity. Polyps can vary widely in size and should be considered a bilateral condition. Rare cases of unilateral polyps should only be diagnosed. Shikhari Taila, known for its anti-inflammatory and antihistaminic properties, this medicated oil penetrates deep into the fine channels, facilitating Sroto Shodhan (cleansing of the channels).

Automated Whole Slide Imaging for Label-Free Histology Using Photon Absorption Remote Sensing Microscopy.

Pathologists rely on histochemical stains to impart contrast in thin translucent tissue samples, revealing tissue features necessary for identifying pathological conditions. However, the chemical labeling process is destructive and often irreversible or challenging to undo, imposing practical limits on the number of stains that can be applied to the same tissue section. Here we present an automated label-free whole slide scanner using a PARS microscope designed for imaging thin, transmissible samples. Peak SNR and in-focus acquisitions are achieved across entire tissue sections using the scattering signal from the PARS detection beam to measure the optimal focal plane. Whole slide images (WSI) are seamlessly stitched together using a custom contrast leveling algorithm. Identical tissue sections are subsequently H&E stained and brightfield imaged. The one-to-one WSIs from both modalities are visually and quantitatively compared. PARS WSIs are presented at standard 40x magnification in malignant human breast and skin samples. We show correspondence of subcellular diagnostic details in both PARS and H&E WSIs and demonstrate virtual H&E staining of an entire PARS WSI. The one-to-one WSI from both modalities show quantitative similarity in nuclear features and structural information. PARS WSIs are compatible with existing digital pathology tools, and samples remain suitable for histochemical, immunohistochemical, and other staining techniques. This work is a critical advance for integrating label-free optical methods into standard histopathology workflows.

Mapping protein-RNA binding in plants with individual-nucleotide-resolution UV cross-linking and immunoprecipitation (plant iCLIP2).

Despite crucial roles of RNA-binding proteins (RBPs) in plant physiology and development, methods for determining their transcriptome-wide binding landscape are less developed than those used in other model organisms. Cross-linking and immunoprecipitation (CLIP) methods (based on UV-mediated generation of covalent bonds between RNAs and cognate RBPs in vivo, purification of the cross-linked complexes and identification of the co-purified RNAs by high-throughput sequencing) have been applied mainly in mammalian cells growing in monolayers or in translucent tissue. We have developed plant iCLIP2, an efficient protocol for performing individual-nucleotide-resolution CLIP (iCLIP) in plants, tailored to overcome the experimental hurdles posed by plant tissue. We optimized the UV dosage to efficiently cross-link RNA and proteins in plants and expressed epitope-tagged RBPs under the control of their native promoters in loss-of-function mutants. We select epitopes for which nanobodies are available, allowing stringent conditions for immunopurification of the RNA-protein complexes to be established. To overcome the inherently high RNase content of plant cells, RNase inhibitors are added and the limited RNA fragmentation step is modified. We combine the optimized isolation of RBP-bound RNAs with iCLIP2, a streamlined protocol that greatly enhances the efficiency of library preparation for high-throughput sequencing. Plant researchers with experience in molecular biology and handling of RNA can complete this iCLIP2 protocol in ~5 d. Finally, we describe a bioinformatics workflow to determine targets of Arabidopsis RBPs from iCLIP data, covering all steps from downloading sequencing reads to identifying cross-linking events ( https://github.com/malewins/Plant-iCLIPseq ), and present the R/Bioconductor package BindingSiteFinder to extract reproducible binding sites ( https://bioconductor.org/packages/release/bioc/html/BindingSiteFinder.html ).

Increasing Lignification in Translucent Disorder Aril of Mangosteen Related to the ROS Defensive Function

Mangosteen fruit has a high potential on the global fruit market, but some disorders, including translucent flesh, are major problems of fruit quality, limiting the marketability. The present study was conducted to compare physiological changes of reactive oxygen species (ROS), cellular lignification between translucent and normal aril, and elucidate the relation. Mangosteen fruits at purple peel color were collected from eastern Thailand during the middle of the rainy season of 2019. Translucent aril accumulated higher lignin content in the tissues, expressing firmer texture ten times higher than normal aril. Lignification was increased in translucent aril by 740% and 25% higher coniferyl alcohol dehydrogenase (CAD) and peroxidase (POD) activity, respectively, induced by high H2O2. Healthy aril performed higher activities of superoxide dismutase (SOD) (8.5 times) and ascorbate peroxidase (APX) (1.3 times) to those in translucent aril. Furthermore, the higher flavonoid content, ascorbic acid content, and antioxidant capacities detected in normal aril could significantly reduce oxidative stress. Although containing high antioxidant systems, healthy aril was found to accumulate higher malonaldehyde content (MDA). This study provides intensive evidence of oxidative stress and the defensive systems between normal and translucent tissues.

Bernotti V-Y plasty techniq ue for root coverage procedure

Gingival recession is the exposure of the root surface resulting from migration of the gingival margin apical to the cementoenamel junction (CEJ). It may be localized or generalized and can be associated with one or more tooth surfaces. Even though gingival recession may occur without any symptoms it can give rise to pain from exposed dentin, patient concern about loss of the tooth, poor esthetics or root caries. The denuded root surfaces cause deterioration in the esthetic appearance, dentin hypersensitivity, and inability to perform proper oral hygiene procedures. The Bernotti V-Y flap is a coronally advanced flap technique with a “V-Y’’ flap design, which consist of a microsurgery technique with or without connective tissue graft, that will improve mucosal biotype and keratinized band in a predictable way for isolated mucosal recession, dehiscence and translucent tissue at teeth or implant sites. It is a conservative flap design, inspired by Semilunar and Modified Semilunar Tarnow’s technique, to minimize interproximal crestal bone loss and possible loss of papillae around teeth or implants, marginal mucosal position is stabilized avoiding adverse aesthetic outcomes. Miller’s Class I and II recession defects with a minimum height of (two) 2 mm are suitable to be selected to perform the Bernotti V-Y flap. The Bernotti V-Y flap consist of a pedicle flap, making a ¨V ¨incision apically to the recession defect and the flap is coronally advanced. Stability and aesthetic outcomes are achieved with the Bernotti V-Y flap.

Meroanencephaly in an English Pointer Neonate

Background: Malformations are structural or functional abnormalities in the organs and structures present at birth. These conditions are rarely described in the newborns of dogs and can lead to their death. Meroanencephaly is a defect of the neural tube closure malformation. This study aims to characterize the clinical-pathological aspects of neonatal meroanencephaly since brain malformations are rare in newborn dogs.Case: A two-day-old English Pointer canine was sent for a necropsy. The newborn belonged to a litter of eight puppies, and only this one had macroscopic cranial alterations. Another puppy that died as a consequence of being trampled by the bitch was also necropsied. The newborn was alive for 48 h until death and presented apathy, crying, sucking reflex and opisthotonus. Macroscopic examination of the baby revealed flattening of the skull, with a slit at the site of bone symphysis fusion, and a slit in the skin of the parietal region, covered by thin, translucent meningeal tissue. The heads of the two animals were examined by radiography to identify the features of anencephaly in one of the animals by visualizing skull bone flattening. Upon removing the skin and exposing the cranial cavity, an irregular reddish mass was revealed, that corresponded microscopically to area cerebrovasculosa, composed of neurons, rudimentary glial tissue, vascular neoformations, as well as hemorrhagic areas.Discussion: Meroanencephaly is a type of anencephaly, a congenital malformation originating from abnormal neurulation, which results from the absence of neural fold fusion during neural tube formation. Live-born anencephalics have some brainstem-driven functions such as spontaneous breathing and some reflex responses, for instance, suction. Several genetic or teratogenic factors, such as viruses, radiation, drugs administered during pregnancy and other pathologies transmitted from bitch to fetus, can lead to this defect. The reports on anatomopathological findings and the presence of area cerebrovasculosa are rare.

A Clearing Technique to Enhance Endogenous Fluorophores in Skin and Soft Tissue

Fluorescent proteins are used extensively in transgenic animal models to label and study specific cell and tissue types. Expression of these proteins can be imaged and analyzed using fluorescent and confocal microscopy. Conventional confocal microscopes cannot penetrate through tissue more than 4–6 μm thick. Tissue clearing procedures overcome this challenge by rendering thick specimens into translucent tissue. However, most tissue clearing techniques do not satisfactorily preserve expression of endogenous fluorophores. Using simple adjustments to the BABB (Benzoic Acid Benzyl Benzoate) clearing methodology, preservation of fluorophore expression can be maintained. Modified BABB tissue clearing is a reliable technique to clear skin and soft tissue specimens for the study of dermal biology, wound healing and fibrotic pathologies.

Measuring Phloem Transport Velocity on a Tissue Level Using a Phloem-Mobile Dye.

Here we describe an in vivo dye-tracking method for measuring phloem transport velocity in seedlings, leaves and petioles and potentially other translucent plant tissues. The method requires measurement of the fluorescent signal of a phloem-mobile fluorescent dye using sensitive photo-sensors placed external to the plant. Following dye application, velocity is determined using laser fluorescence bleaching and measuring the time it takes for the bleach front to reach a light sensor. This method can be used to measure phloem transport velocity on intact plants with minimal disturbance and has a potential to be used under a variety of growth conditions and in the field. Because there are large differences among species in their anatomy, this method should be optimized to individual plants and tissue types.

Scleria cheekii, a new species of Scleria subgenus Hypoporum (Cyperaceae, Cyperoideae, Sclerieae) from Cameroon

An endemic species of Scleria (Cyperaceae) from Cameroon, Scleria cheekii Bauters, is described as new to science. Scleria cheekii is an annual known from basalt pavements in the Bamenda Highlands in Cameroon. It can be distinguished from related species by the presence of tuberculated yellow-orange translucent tissue covering the trigonous stipe. This species is described, illustrated and compared with morphologically closely allied species.

Analytical prediction of sub–surface thermal history in translucent tissue phantoms during plasmonic photo–thermotherapy (PPTT)

Knowledge of thermal history and/or distribution in biological tissues during laser based hyperthermia is essential to achieve necrosis of tumour/carcinoma cells. A semi–analytical model to predict sub–surface thermal distribution in translucent, soft, tissue mimics has been proposed. The model can accurately predict the spatio–temporal temperature variations along depth and the anomalous thermal behaviour in such media, viz. occurrence of sub-surface temperature peaks. Based on optical and thermal properties, the augmented temperature and shift of the peak positions in case of gold nanostructure mediated tissue phantom hyperthermia can be predicted. Employing inverse approach, the absorption coefficient of nano-graphene infused tissue mimics is determined from the peak temperature and found to provide appreciably accurate predictions along depth. Furthermore, a simplistic, dimensionally consistent correlation to theoretically determine the position of the peak in such media is proposed and found to be consistent with experiments and computations. The model shows promise in predicting thermal distribution induced by lasers in tissues and deduction of therapeutic hyperthermia parameters, thereby assisting clinical procedures by providing a priori estimates.

<p class="MsoTitle"><span lang="EN-US">Scleria pantadenia</span><span lang="EN-US"> and Scleria tricristata: Two new species of Scleria subgenus </span><span lang="EN-US">Hypoporum</span><span lang="EN-US"> (Cyperaceae, Cyperoideae, Sclerieae) from Tanzania</span><p class="MsoNormal">

Scleria pantadenia Meganck & Bauters and Scleria tricristata Meganck & Bauters are presented as new species of Scleria (Cyperaceae) from Tanzania. Scleria pantadenia is a small, tufted annual sedge species characterized by its delicate, brittle appearance and its small nutlets with gland-like tubercules and basal rim of swollen tuberculate cells. Scleria tricristata is a small, tufted annual sedge species characterized by a single, terminal unbranched spike and its remarkable nutlets with three protruding dentate ridges of translucent tissue. Both species are described, illustrated and compared with their closest relatives.

Confocal Laser Endomicroscopy and NSAID Enteropathy: Where Next?

Although Marvin Minsky invented the confocal microscope in 1957, the development of the personal computer in the early 1980s was needed before this invention was widely used in the biological sciences, with commercial confocal endoscopes not available until the first years of the twenty-first century. Confocal microscopy is a method of obtaining sharp images of fluorescence from within solid translucent tissue. Underlying this technique is the fundamental physical observation that when bright light of a particular wavelength (the excitation wavelength) strikes a fluorescent molecule, light of a longer wavelength (the emission wavelength) is emitted. In a confocal microscope, a series of mirrors sweeps a beam of excitation laser light across the tissue in order to restrict excitation to a specific location at any one time. In response to this excitation, fluorescent light emits from the entire depth of the tissue, producing a blurred image. To obtain a sharp image from a single plane within the tissue, a pinhole is positioned in front of the fluorescence detector so that only light from the focal plane within the tissue, termed the confocal point, is transmitted, such that light from above and below the focal plane is excluded. The beauty of this technique is that no physical slicing of the tissue is required as in conventional histology. Instead, the optical sectioning of the confocal microscope creates sharp images from deep within solid tissue. When incorporated into an endoscope, microscopic fluorescent images of the gut mucosa are obtained from beneath the surface. In principle, such images when obtained in loosely structured organs such as the liver can be acquired at depths of 200‐300 lm, although in the intestine the maximum depth is \50 lm due to absorption and diffraction of the emitted light by epithelial nuclei. To date, confocal endomicroscopy (CLE) has been used in cancer surveillance in inflammatory bowel disease and in Barrett’s esophagus due to its ability to target endo

Translucent Scale and Leathery Scale Disorders of Onion Bulbs: Describing Symptoms and Causes

In Northern Europe, onion (Allium cepa L.) is grown from sets planted out in early May and lifted in early September. After a field-curing period the bulbs are put into storage and dried at 25–30°C until the neck is completely dry before the temperature is gradually reduced to 0°C for long-term storage. Watery scale (translucent and leathery scale) is a postharvest problem in our area causing loss of yield and quality. Watery scale includes the following categories of disorders: leathery scale, which appears as thick, dark scales between outer dry and inner fleshy scales, and translucent scale, which appears as glassy, firm fleshy scales further into the bulb. Symptoms of watery scale were clarified based on visual appearance, tissue pH, electrical conductivity, and contents of ethanol, lactic acid, and acetic acid. The disorders were linked to disturbances in gas exchange of bulbs with high internal CO2 and/or low internal O2 levels. Bulbs were exposed to handling treatments leading to increased respiration and/or reduced gas exchange from the interior of bulbs to the outside atmosphere. Internal bulb atmosphere, gas flux, and gas permeability (of O2) of onion scale and tissues were measured to explain why damage occurred. Translucent scale generally occurred in the middle and upper parts of the outer two fleshy scales but could include the entire bulb. A clear reduction in electrical conductivity of translucent tissue was measured, indicating leaching. A rise in pH occurred in translucent scales with weak symptoms followed by a reduction in pH. Leathery scale tissues had a pH around 3.5 and a rise in ethanol content, indicating anaerobic respiration. Dry scale and epidermis of fleshy scale had very low oxygen permeability. Transport of gas through the neck region was 10–20 times higher than transport through the scale. Gas flux was related to need of O2 in respiring tissue and to problems of adverse internal atmosphere and development of disordered scales. Normal cultivation of onion and handling of bulbs resulted in low occurrence of disordered bulbs. Weak symptoms of translucent scale could disappear in storage, but usually symptoms became more severe after long-term storage. Dropping and pressure treatments produced more translucent scale but had no effect on leathery scale. Internal CO2 concentration, as high as 22.3%, occurred after combined dropping and pressure treatments. The results explained the induction of translucent scale. Leathery scale was not affected by the treatments examined in this study. Further work is needed to understand why leathery scale develops and how it can be minimized.

Cellular Mechanisms of Aneurysm Occlusion after Treatment with a Flow Diverter

To characterize the progression of healing across aneurysm necks following treatment with a flow diverter in a rabbit aneurysm model. With institutional animal care and use committee approval, saccular aneurysms were created in 20 rabbits and treated with flow diverters. On days 1, 3, and 7 and weeks 4 and 8 after implantation, the aneurysm and the device-implanted vessel were harvested. En face staining of the gross specimen was performed for endothelial cells, endothelial progenitor cells, smooth muscle cells, and inflammatory cells. The parent artery segments covered by the flow diverters were completely devoid of endothelial cells at 1 and 3 days but had completely reendothelialized by 7 days. At all time points, the struts along the patent portions of the aneurysm necks harbored scattered tissue islands composed exclusively of inflammatory cells. At 4 and 8 weeks, all samples contiguous with the tissue along the parent arteries had translucent tissue present along the occluded segments of the aneurysm neck. The vast majority of endothelial cells were contiguous with the parent artery and had smooth muscle cells underlying them. Endothelial progenitor cells were not observed along the neck of any aneurysm. Aneurysm closure was noted only when complete or nearly complete endothelialization over the device struts was present. The initial event following flow diversion treatment is adherence of clusters of inflammatory cells across the aneurysm neck. Endothelialization is relatively delayed and derived exclusively from cells in the adjacent parent artery.

Translucent tissue defect in potato (Solanum tuberosum L.) tubers is associated with oxidative stress accompanying an accelerated aging phenotype

Translucent tissue defect (TTD) is an undesirable postharvest disorder of potato tubers characterized by the development of random pockets of semi-transparent tissue containing high concentrations of reducing sugars. Translucent areas turn dark during frying due to the Maillard reaction. The newly released cultivar, Premier Russet, is highly resistant to low temperature sweetening, but susceptible to TTD. Symptoms appeared as early as 170 days after harvest and worsened with time in storage (4-9 °C, 95 % RH). In addition to higher concentrations of glucose, fructose and sucrose, TTD resulted in lower dry matter, higher specific activities of starch phosphorylase and glc-6-phosphate dehydrogenase, higher protease activity, loss of protein, and increased concentrations of free amino acids (esp. asparagine and glutamine). The mechanism of TTD is unknown; however, the disorder has similarities with the irreversible senescent sweetening that occurs in tubers during long-term storage, where much of the decline in quality is a consequence of progressive increases in oxidative stress with advancing age. The respiration rate of non-TTD 'Premier Russet' tubers was inherently higher (ca. 40 %) than that of 'Russet Burbank' tubers (a non-TTD cultivar). Moreover, translucent tissue from 'Premier Russet' tubers had a 1.9-fold higher respiration rate than the average of non-translucent tissue and tissue from non-TTD tubers. Peroxidation of membrane lipids during TTD development resulted in increased levels of malondialdehyde and likely contributed to a measurable increase in membrane permeability. Superoxide dismutase and catalase activities and the ratio of oxidized to total glutathione were substantially higher in translucent tissue. TTD tubers also contained twofold less ascorbate than non-TTD tubers. TTD appears to be a consequence of oxidative stress associated with accelerated aging of 'Premier Russet' tubers.

Evaluation of human sclera after femtosecond laser ablation using two photon and confocal microscopy

Glaucoma is the second-leading cause of blindness worldwide and is often associated with elevated intraocular pressure (IOP). Partial thickness intrascleral channels can be created with a femtosecond laser operating at a wavelength of 1700 nm. Such channels have the potential to increase outflow facility and reduce elevated IOP. Analysis of the dimensions and location of these channels is important in understanding their effects. We describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in human cadaver eyes. High-resolution images, hundreds of microns deep in the sclera, were obtained to allow determination of the shape and dimension of such channels. This demonstrates that concept of integrating femtosecond laser surgery, and two-photon and confocal imaging has the future potential for image-guided high-precision surgery in transparent and translucent tissue.

Implantation of newborn mice skin cells with chamber method to construct a model of hair follicle development

To construct a convenient, reliable and visual model of hair follicle development to test the hair-inductive potential of follicular cells and investigate the molecular mechanism regulating hair follicle morphogenesis and cycling. An open chamber was transplanted into the nude mice dorsal skin, dermal and epidermal cells isolated from newborn C57BL/6 mice skin were mixed at a specific ratio and then injected into the chamber together, 1 week after transplantation, the chamber was removed, and then, hair formation and regeneration after hair plucking was observed. 1 week after cells implantation, the wound was moist without apparent contraction and among that pink and translucent tissue was formed. 2 weeks after implantation, the wound healed completely. 3 weeks after implantation, black hair grew from the skin was observed. 4 weeks after implantation, thick and black hair grew from the skin vertically. Completely developed structure of hair follicle was observed with paraffin section and HE staining. 1 week after plucking, new hair had regrown. The ratio of cell component was varied, whereas the other component was fixed at 1 x 10(7) cells. When the number of epidermal cells was reduced to 1 x 10(6) cells, the efficiency of hair follicle reconstitution was mostly unchanged. On the other hand, the density of newly formed hair was diminished considerably by reducing the number of dermal cells to 5 x 10(6) cells or lower. Neither epidermal cells nor dermal cells transplanted alone formed hair follicle. Newborn mice skin cells transplanted by chamber method can construct a complete model of hair follicle development, which can be used to test the hair-inductive potential of follicular cells and investigate the molecular mechanism regulating hair follicle morphogenesis and cycling.

Cystoid changes within astrocytic hamartomas of the retina in tuberous sclerosis.

The purpose of this study was to report two unusual cases of cavitations within astrocytic hamartoma, a feature that causes the tumor to appear cystic. To our knowledge, this is the first report on cavitary astrocytic hamartoma. In this observational case report, we identify two cases of cavitary astrocytic hamartoma imaged recently with spectral domain-optical coherence tomography. In both of our patients, the cavitary portion was visible clinically as an empty cavity surrounded by translucent tissue. Optical coherence tomography examination of both cases confirmed the presence of a cavity surrounded by tumor tissue. In these two patients, optical coherence tomography showed a new feature of cavitation within retinal astrocytic hamartoma.

Intravascular ultrasound, angioscopic and histopathological characterisation of heterogeneous patterns of restenosis after sirolimus-eluting stent implantation: insights into potential “thromborestenosis” phenomenon

Histopathology of DES restenosis remains unclear. The purpose is to examine restenotic tissue characteristics after sirolimus-eluting stent (SES) and comparing with that after bare-metal stent (BMS). Intravascular ultrasound (IVUS), coronary angioscopy (CAS), and directional coronary atherectomy were performed simultaneously in 21 patients who presented restenosis after SES (n=13) and BMS (n=8). Mean time of restenosis was 10.8 months in the SES versus 7.5 months in the BMS. Typical "black hole'', echolucent appearance by IVUS was observed in one SES case, and corresponded to a fibrin rich tissue by histology which appeared translucent tissue by CAS. CAS did not reveal red thrombus, but showed white thrombus in six SES versus two BMS (46.2% vs. 25.0%, p=0.597). Histology demonstrated various patterns after SES including thrombus, fibrin, inflammatory infiltrate, and collagen-matrix rich tissue, while thrombus component was not detected in BMS. Thrombus and fibrin deposition detected by either CAS or histopathology were observed more frequently in SES than in BMS group (92.3% vs. 25.0%, p=0.007). Restenosis after SES and BMS have different clinical and histological patterns. SES restenosis may be frequently associated with thrombus component.

Novel intermediate-term individual marking technique for slugs

To determine the lowest dose required preventing elastomer rejection, we added 0.025, 0.05, 0.75 or 1.0 mg of tricaine methanesulphonate per 50 ml water buffered with 1 ml liquid sodium bicarbonate. The bath was approximately 5 cm deep, which was shallow enough to ensure that the pneumostome was clear of the liquid. A pipette was used continually to bathe the slug in the solution. We used five slugs for each trialgroup. In addition to these 20 slugs, we included a control group of ten slugs. The control group was not subjected to anaesthetic or marking treatment, but was placed in water buffered with 1 ml liquid sodium bicarbonate. All animals were randomly assigned to either the control or anaesthetic treatment. They were either placed in the buffered control or anaesthetizing treatment bath for 15 min at 228C. All doses of anaesthetic resulted in the animals relaxing without causing excessive mucus production. Preliminary tests were conducted to find suitable marking sites on the slugs. We attempted to inject the elastomer beneath the skin on the mantle; however, the skin in this area tended to tear easily. Further, if the injection was too deep it could puncture the body wall, potentially resulting in mortality. Our preliminary trials found that that the elastomer could be easily and consistently applied to the translucent tissue of the foot, with minimal skin tearing and high visibility. Immediately following anaesthesia, we implanted visible elastomers (VIEs; Northwest Marine Technology, Shaw Island, Washington) into the 20 relaxed slugs. The elastomer was prepared by mixing 10 parts of the coloured component with 1 part curing agent as directed by Northwest Marine Technology. The elastomer-filled needle was inserted at a low angle into the translucent tissue on the outside edge of the foot. A very small amount of elastomer was then injected, resulting in