The three-dimensional findings of the surface and from a cross section from a case of disseminated superficial porokeratois using scanning electron microscopy are reported. On the surface of the skin, irregular keratin with a serpiginous distribution was seen. A gross aspect of keratin in the hyperkeratotic wall was also observed and compared to the normal area, in which the release of corneocytes seemed normal. The cross-sectional imaging easily identified the cornoid lamella, with compact keratin surrounded by normal stratum corneum.

Tinea nigra is a rare superficial mycosis caused by Hortaea werneckii. This infection presents as asymptomatic brown to black maculae mostly in palmo-plantar regions. We performed scanning electron microscopy of a superficial shaving of a tinea nigra lesion. The examination of the outer surface of the sample showed the epidermis with corneocytes and hyphae and elimination of fungal filaments. The inner surface of the sample showed important aggregation of hyphae among keratinocytes, which formed small fungal colonies. The ultrastructural findings correlated with those of dermoscopic examination - the small fungal aggregations may be the dark spicules seen on dermoscopy - and also allowed to document the mode of dissemination of tinea nigra, showing how hyphae are eliminated on the surface of the lesion.

The purpose of this study is to compare scanning electron microscopy findings of the blister roof in three distinct bullous diseases: one intraepidermal acantholytic (pemphigus foliaceus); one due to hemidesmosomal dysfunction (bullous pemphigoid); and one secondary to anchoring fibril dysfunction - type VII collagen (dystrophic epidermolysis bullosa). In pemphigus foliaceus, acantholytic phenomena were readily demonstrated. In bullous pemphigoid, the epidermis had a solid aspect. In dystrophic epidermolysis bullosa a net was seen in the blister roof.

Na epidermólise bolhosa distrófica, o defeito genético das fibrilas ancorantes leva à clivagem abaixo da membrana basal, com sua consequente perda. Realizamos microscopia eletrônica de varredura do teto invertido de uma bolha de um caso de epidermólise bolhosa distrófica, cujo diagnóstico foi confirmado com imunomapeamento e com sequenciamento gênico. Com uma ampliação de 2.000 vezes, pôde ser facilmente identificada uma rede ligada ao teto da bolha. Essa rede era composta por fibras achatadas e entrelaçadas. Com grandes aumentos, fibras de diferentes tamanhos puderam ser observadas: algumas finas, medindo cerca de 80 nm, e outras mais largas, medindo entre 200 nm e 300 nm.

In dystrophic epidermolysis bullosa the genetic defect of anchoring fibrils leads to cleavage beneath the basement membrane, with its consequent loss. We performed scanning electron microscopy of an inverted blister roof of a case of dystrophic epidermolysis bullosa, confirmed by immunomapping and gene sequencing. With a magnification of 2000 times a net attached to the blister roof could be easily identified. This net was composed of intertwined flat fibers. With higher magnifications, different fiber sizes could be observed, some thin fibers measuring around 80 nm and thicker ones measuring between 200 and 300 nm.

Realizamos microscopia eletrônica de varredura do teto invertido de uma bolha de um caso de pênfigo foliáceo. Com pequeno aumento, a perda da adesão intercelular pôde ser vista claramente. Os queratinócitos acantolíticos demostraram um contorno irregular, algumas vezes poligonal. Células arredondadas, como vistas tipicamente na microscopia óptica, também foram observadas. O exame de um teto de bolha permite uma documentação ultraestrutural das alterações acantolíticas.

We performed scanning electron microscopy of an inverted blister roof in a case of pemphigus foliaceus. The loss of intercellular adherence could be easily seen with low magnification. The acantholytic keratinocytes displayed an irregular and sometimes polygonal contour. Round cells, typically seen in light microscopy, were also observed. The examination of a blister roof allows ultrastructural documentation of the acantholytic changes.

O líquen escleroso é uma afecção inflamatória caracterizada por placas esbranquiçadas fibróticas ocorrendo preferentemente na pele genital. Realizamos exame com microscópio eletrônico de varredura da derme de uma lesão de líquen escleroso. Ao exame as fibras colágenas normais puderam ser facilmente identificadas, bem como a transição para a área alterada, a qual aparece homogeneizada. Nessa área as fibras parecem aderidas umas às outras e com aspecto em colar de pérolas. Na derme superficial essa homogeneização é ainda mais evidente, com as fibras bem aderidas e ainda com as estruturas arredondadas. Ruptura das fibras e células inflamatórias não foram observadas. Os fenômenos autoimunes que ocorrem na matriz extracelular nessa enfermidade devem levar à agregação de imunocomplexos e/ou proteínas alteradas nas fibras colágenas, por essa razão elas aparecem hialinizadas na microscopia óptica.

Lichen sclerosus is an acquired inflammatory condition characterized by whitish fibrotic plaques, with a predilection for the genital skin. We performed scanning electron microscopy of the dermis from a lesion of lichen sclerosus. Normal collagen fibers could be easily found in deeper layers of the specimen, as well as the transition to pathologic area, which seems homogenized. With higher magnifications in this transitional area collagen fibers are adherent to each other, and with very high magnifications a pearl chain aspect became evident along the collagen fibers. In the superficial dermis this homogenization is even more evident, collagen fibers are packed together and round structures are also observed. Rupture of collagen fibers and inflammatory cells were not found. These autoimmune changes of the extracellular matrix lead to the aggregation of immune complexes and/or changed matrix proteins along the collagen fibers, the reason why they seem hyalinized when examined by light microscopy.

A epidermólise bolhosa albo-papulóide de Pasini é uma variante rara da forma generalizada de epidermólise bolhosa distrófica dominante. Uma paciente de 30 anos apresenta desde a infância pápulas e bolhas disseminadas. A microscopia óptica de uma bolha demonstrou clivagem dermo-epidérmica; além disso áreas focais de desprendimento dermo-epidérmico foram encontradas. A microscopia eletrônica de transmissão identificou a clivagem abaixo da lâmina densa. É importante que se reconheça essa variante de epidermólise bolhosa, já que o aspecto clínico predominante são pápulas disseminadas e não bolhas como nas outras formas de epidermólise bolhosa.

Pasini's albopapuloid epidermolysis bullosa is a very rare subtype of generalized dystrophic dominant epidermolyis bullosa. A 30 year-old white female patient presented since her childhood disseminated small blisters and papules. Light microscopy of a blister showed dermal-epidermal cleavage; moreover, focal areas of dermal-epidermal splitting were also observed. Transmission electron microscopy also identified focal areas of cleavage, which were seen below the lamina densa. It is important to recognize this condition as a variant of epidermolysis bullosa, since the most important cutaneous findings are generalized papules and not blisters and erosions as in other forms of epidermolysis bullosa.

Na epidermólise bolhosa distrófica, o defeito genético das fibrilas de ancoragem leva à clivagem abaixo da membrana basal com sua consequente perda. Uma paciente de 46 anos apresentava bolhas pré-tibiais associadas à distrofia ungueal. Seus dois filhos apresentavam hipo e anoníquia, afetando todas as unhas dos pododáctilos e dos primeiros, segundos e terceiros quirodáctilos. O sequenciamento de DNA identificou no exon 75 do gene COL7A1 uma mutação patológica: c.6235G>A (p.Gly2079Arg). O imunomapeamento identificou o colágeno IV no teto e colágeno VII no assoalho de uma bolha , confirmando o diagnóstico de epidermólise bolhosa distrófica. A microscopia eletrônica de varredura de um teto invertido de bolha demonstrou rede de colágeno aderida ao mesmo. A variabilidade clínica encontrada nessa família já foi escrita e confirma, que o subtipo ungueal das epidermólises bolhosas é uma forma distrófica.

In dystrophic epidermolysis bullosa, the genetic defect of anchoring fibrils leads to cleavage beneath the basement membrane and its consequent loss. A 46 year-old female patient presented blisters with a pretibial distribution associated with nail dystrophy. Her two children had hyponychia and anonychia, which affected all toe nails and the thumb, forefinger and middle finger. DNA sequencing identified in exon 75 of COL7A1 gene a pathologic mutation: c.6235G>A (p.Gly2079Arg). Immunomapping of a blister demonstrated collagen IV (basal membrane) in the blister roof and collagen VII in its floor, confirming dystrophic epidermolysis bullosa. Scanning electron microscopy of an inverted blister showed net-forming collagen attached to the blister roof . The variability found in this family has already been reported and confirms, on a clinical basis, the nail subtype as a dystrophic variant.

Pili canaliculi é alteração capilar rara, geralmente herdada de forma autossômica dominante, caracterizada por cabelos arrepiados, pertencendo ao espectro dos cabelos impenteáveis. Poucos estudos encontram ou explicitam sua variabilidade clínica. Uma família com 12 indivíduos afetados foi estudada, e o acometimento do couro cabeludo demonstrou grande variabilidade, desde cabelos arrepiados, difíceis de pentear, hipotricose leve ou intensa, até atriquia adquirida. O exame de microscopia óptica de cortes transversais dos cabelos, a estereomicroscopia e a microscopia eletrônica de varredura confirmaram o diagnóstico, demonstrando canais na superfície dos cabelos.

Pili canaliculi is rare hair disorder, usually inherited as an autosomal dominant trait, clinically characterized by uncombable hair. Few studies report its clinical variability. A family with 12 affected individuals was examined and involvement of hairs ranged from uncombable hairs, mild or intense hypotrichosis to acquired atrichia. Optical microscopy of hair cross sections, stereomicroscopy and scanning electron microscopy confirmed the diagnosis and showed hair shafts with characteristic longitudinal grooves.

OBJECTIVE: To examine the epidermis in induced phytophotodermatitis using transmission electron microscopy in order to detect histologic changes even before lesions are visible by light microscopy. INTRODUCTION: In the first six hours after the experimental induction of phytophotodermatitis, no changes are detectable by light microscopy. Only after 24 hours can keratinocyte necrosis and epidermal vacuolization be detected histologically, and blisters form by 48 hours. METHODS: The dorsum of four adult rats (Rattus norvegicus) was manually epilated. After painting the right half of the rat with the peel juice of Tahiti lemon, they were exposed to sunlight for eight minutes under general anesthesia. The left side was used as the control and exposed to sunlight only. Biopsies were performed immediately after photoinduction and one and two hours later, and the tissue was analyzed by transmission electron microscopy. RESULTS: No histological changes were seen on the control side. Immediately after induction, vacuolization in keratinocytes was observed. After one hour, desmosomal changes were also observed in addition to vacuolization. Keratin filaments were not attached to the desmosomal plaque. Free desmosomes and membrane ruptures were also seen. At two hours after induction, similar changes were found, and granular degeneration of keratin was also observed. DISCUSSION: The interaction of sunlight and psoralens generates a photoproduct that damages keratinocyte proteins, leading to keratinocyte necrosis and blister formation. CONCLUSIONS: Transmission electron microscopy can detect vacuolization, lesions of the membrane, and desmosomes in the first two hours after experimental induction of phytophotodermatitis.

A estrutura mais superficial da epiderme dos frutos é constituída por fina camada de cera, responsável por mecanismos de proteção, principalmente relacionados à evapotranspiração e ao ataque de patógenos. A visualização de cera por microscopia eletrônica de varredura apresenta dificuldades no processamento da amostra, pois os métodos rotineiramente utilizados ocasionam sérios danos a esta camada ou, simplesmente, provocam sua eliminação devido ao processo de solubilização causado pelos reagentes utilizados. Alguns autores têm utilizado temperaturas baixas (-90ºC) para o processamento de amostras. No Brasil, tentativas de observação de cera em maçãs, utilizando a microscopia eletrônica de varredura, têm sido realizadas; entretanto, os resultados não foram satisfatórios. No Laboratório de Microscopia Eletrônica da Embrapa Clima Temperado, foi adaptado um método simples e que tem apresentado bons resultados. Para este experimento, foram retiradas pequenas porções da casca de maçãs da região equatorial dos frutos. As amostras foram fixadas em lâminas histológicas, utilizando fita adesiva nas extremidades. Posteriormente, foram colocadas em dessecador contendo sílica gel, para que ocorresse a desidratação. Após 72 horas, foram retirados fragmentos para fixação em stubs e metalização com ouro. Foi usado um microscópio eletrônico ZEISS (DSM-940A), regulado à distância de trabalho de 15mm, voltagem de aceleração de 10 kV e ampliação de 3.000 X. As imagens obtidas permitiram avaliar a interferência dos fatores ambientais utilizados no processo de frigoconservação de maçãs.

The fruit epidermis is covered by a thin layer of wax which gives protection to the fruit, mainly against water loss and pathogenic organisms. Several studies using scanning electron microscope have been developed to study that wax. The wax layer visualization through the scanning microscope is difficult due to some problems of sample preparation. The usual procedures cause serious damages to the wax layer or even eliminate it because of its solubilization by some of the reagents. Several researchers have kept the samples at very low temperature (-90ºC) during the preparation. Wax observations in Brazil by using electronic scanning microscope have been performed, however in some of them results were not consistent. A relatively simple procedure at the Laboratory of Electron Microscopy at Embrapa Temperate Climate has been tried to overcome such difficulties. Such procedure consists of collecting a small piece of the fruit epidermis (1cm²) taken from the equatorial part of the apples. After sampling it the piece of epidermis is placed on a glass slide hold by the extremes with a sticky tape. The sample on glass slide is kept for 72 hours into a dessicator containing drierite for the sample dehydration. Then, small pieces of the sample (2 mm²) were taken and hold on stubs followed by metalization in gold. Following that, the sample is observed using a Zeiss (DSM - 940A) electronic microscope at a working distance of 15 mm and acceleration voltage of 10 KV. The obtained images made possible to evaluate the action of some experimental treatments which have been used on apples during cold storage.