Preparación de hemisferios cerebrales para disección de tractos / Preparation of cerebral hemispheres for tracts dissection

Desde la antigüedad se han desarrollado técnicas para el estudio del cerebro con fines didácticos o neuroquirúrgicos. Hacia 1934 Josef Klingler desarrolla una técnica de preparación de hemisferios cerebrales que basada en la fijación con formalina y el congelamiento para aislar los tractos cerebrales. El objetivo de la presente artículo ha sido analizar los métodos de preparación utilizados para la disección de tractos en cerebros humanos y de animales. Se realizó una revisión de la literatura en las bases de datos Web of Science, Scopus, Pubmed, Medline y Scielo, utilizando como descriptores: Disección, Cerebro, Tracto, con el operador booleano "AND" entre ellos, en los idiomas inglés y español, hasta junio de 2018. Fueron seleccionados 26 documentos, para el análisis se determinaron las variables: espécimen, número de hemisferios cerebrales, concentración de formalina, tiempo de fijación, temperatura, tiempo de congelamiento y tractos identificados. En la literatura seleccionada, un total de 410 hemisferios cerebrales fueron analizados, 372 de humanos y 38 de animales; 330 fueron conservados en formalina al 10 %, 20 en formalina al 5 % y el resto en otras concentraciones. El tiempo de fijación fue variable entre 10 y 180 días, así como la temperatura y tiempo de congelación (-10 ºC y -20 ºC, entre 8 y 30 días). En todos los casos se reportó que, en su totalidad o parcialmente, los fascículos cerebrales de asociación fueron aislados. En la preparación de hemisferios cerebrales para disección de tractos, Ludwig & Klingler (1956) recomiendan que en la fijación de los especímenes se utilice formalina al 5 %, sin embargo, el 80 % de los hemisferios utilizados fueron fijados en formalina al 10%, y en esta concentración, el tiempo de fijación, temperatura y tiempo de congelación fue variable, lográndose, en todos los casos analizados, la disección parcial o total de los tractos.

Since ancient times, techniques for the study of the brain have been developed for didactic or neurosurgical purposes. By 1934, Josef Klingler developed a cerebral hemisphere preparation technique based on formalin fixation and freezing to isolate the cerebral tracts. The aim of this article was to analyze the preparation methods used for tracts dissection in human and animal brains. A review of the literature using Web of Science, Scopus, Pubmed, Medline and Scielo databases, with the following descriptors: Dissection, Brain, Tract, with the boolean operator "AND" among them, also in spanish, until June 2018. Twenty-six documents were selected, and we analized the following variables: specimen, number of cerebral hemispheres, formalin concentration, fixing time, temperature, freezing time and tracts. In the selected literature, a total of 410 cerebral hemispheres were analyzed, 372 from humans and 38 from animals; 330 were preserved in 10 % formalin, 20 in 5 % formalin and the rest in other concentrations. The fixation time was variable between 10 and 180 days, as well as the temperature and freezing time (-10 ºC and -20 ºC, between 8 and 30 days). In all cases it was reported that, in whole or in part, the cerebral fascicles of association were isolated. In the preparation of cerebral hemispheres for dissection of tracts, Klingler recommend that 5 % formalin for the fixation of specimens; however, 80 % of the hemispheres used were fixed in 10 % formalin, and in this concentration, the time of fixation, temperature and time of freezing was variable, achieving, in all the cases analyzed, the partial or total dissection of the tracts.

Correlación cito-colpo-histológica en lesiones premalignas del cuello uterino en el Hospital Básico Píllaro en Ecuador / Cito-Colpo-Histological Correlation in Premalignant Cervical Lesions

Introducción: el cáncer cervicouterino es una de las amenazas más graves para la vida de las mujeres. Actualmente en el mundo lo padecen más de un millón de ellas. En Ecuador, ocupa el segundo lugar en incidencia y causa 1,2 por ciento de muertes anuales en el país. Su detección oportuna es posible gracias a la citología cervicovaginal que contribuye eficazmente a detectar lesiones precancerosas y disminuir significativamente el carcinoma del cuello uterino. El estudio de la paciente se complementa con colposcopia y toma de biopsia para aumentar la certeza diagnóstica. En algunos casos, no se ha observado una buena correlación diagnóstica. Objetivo: determinar la relación citocolpohistológica en pacientes atendidas con Papanicolaou alterado en consulta de Patología del Tracto Genital Inferior. Métodos: se revisaron 82 historias clínicas de pacientes atendidas con Papanicolaou alterado en la consulta de Patología del Tracto Genital Inferior en el Hospital Básico Píllaro de Ecuador desde abril de 2015 hasta abril de 2016. Resultados: del total de pacientes, 32,9 por ciento tenían entre 30 y 39 años de edad; 90,2 por ciento iniciaron sus relaciones sexuales durante la adolescencia. De ellas, 89 por ciento tuvo entre una y cinco parejas sexuales; 59,7 por ciento tuvo entre uno y tres partos. Existió un 21,4 por ciento de correlación cito-colposcópica en el diagnóstico de las lesiones intraepiteliales de bajo grado. La relación colpo-histológica mostró un 87,5 por ciento de coincidencias en las lesiones intraepiteliales de bajo grado y en las lesiones intraepiteliales de alto grado un 71,4 por ciento. Conclusiones: el inicio precoz de la actividad sexual, las múltiples parejas sexuales y la multiparidad continúan resaltando en la aparición de las lesiones premalignas del cuello uterino(AU)

Introduction: Cervical cancer is one of the most serious threats to the lives of women. In the world today, more than a million of them suffer from it. In Ecuador, it ranks second in incidence and causes 1.2 percent of annual deaths in the country. Its timely detection is possible thanks to cervicovaginal cytology that contributes effectively to detecting precancerous lesions and significantly decreasing carcinoma of the cervix. The patient's study is complemented by colposcopy and biopsy to increase diagnostic certainty even when good diagnostic correlation has not been observed in some cases. Objective: Determine the cyto-colpo-histological relationship in patients treated due to altered Papanicolaou, in consultation of Pathology of the Lower Genital Tract in Píllaro Basic Hospital. Ecuador. Methods: In the present study, we reviewed 82 clinical records of patients treated with altered Papanicolaou in the Lower Genital Tract Pathology consultation at the Píllaro Basic Hospital, Ecuador from April 2015 to April 2016. Results: 32.9 percent of patients aged 30 to 39 years; 90.2 percent started sexual intercourse throughout adolescence. 89 percent had one to five sexual partners. 59.7 percent had one to three deliveries. There was 21.4 percent cyto-colposcopic correlation in the diagnosis of low-grade intraepithelial lesions (LSIL). The colpo-histological relationship showed 87.5 percent of coincidences in the LSIL and 71.4 percent. showed high-grade intraepithelial lesions (HSIL)(AU)

Perda de amostras em tissue microarray: comparação entre técnicas com uso de fita adesiva comercial, lâminas silanizadas pelo método tradicional ou por método modificado / Loss of tissue samples in the tissue microarray: comparison between techniques using commercial adhesive tape, silane-coated microslides by the conventional method and by a modified technique

Introdução/objetivo: O microarranjo tecidual, ou tissue microarray (TMA), permite avaliar múltiplas amostrasde tecido em um único bloco. Um dos problemas do TMA é o descolamento dos cortes teciduais, por isso, para reduzir essa perda, tem-se utilizado fita adesiva especial comercial. Não há relatos comparando o uso dessas fitas adesivas com a técnica de silanização modificada. O objetivo desse estudo foi comparar as perdas de cortes entre lâminas usando fitas adesivas comerciais, lâminas silanizadas por técnica convencional elâminas silanizadas por técnica modificada, com menor consumo de acetona. Material e método: O TMA foiconstruído com blocos de tecido hepático, em dispositivo de base fixa, colocando-se 32 cilindros de 2 mmde diâmetro em duplicata e espaçamento de 2,2 mm. Quinze secções de 4 μm foram colocadas em lâminas silanizadas a 4% por técnica convencional (grupo 1), 15 em lâminas silanizadas com técnica modificada (6%de silano e com uso mínimo de acetona) (grupo 2) e 15 em lâminas com fita adesiva comercial de acordo comas recomendações do fabricante (grupo 3). Todas as lâminas foram processadas por imuno-histoquímica para citoqueratina 18, com recuperação antigênica em tampão citrato pH 6, em microondas. As perdas de amostrasforam quantificadas e expressas como: perda total (≥ 80%), quase total (75% a 79%) ou parcial (50% a 74%).Resultados: A perda de tecidos foi semelhante nos três grupos: com silanização tradicional, modificada oufita adesiva comercial (4,9 vs. 3,1 vs. 8,1, respectivamente) (análise de variância [ANOVA], p = 0,3654). Umadas lâminas com a fita adesiva apresentou descolamento artefatual de todos os tecidos e outra de 20 tecidosem um dos lados. Nenhuma das lâminas silanizadas apresentou tal artefato. Conclusão: Lâminas silanizadas têm resultados satisfatórios, requerem menos treinamento técnico e reduzem os custos da utilização do TMA justificando seu uso em pesquisa...

Introduction/objective: The tissue microarray (TMA) technique allows the evaluation of multiple tissue samplesin a single block. One of the problems of TMA is the ungluing of tissue sections, thus commercial adhesive tape has been used to reduce this loss. There are no reports comparing the use of the commercial adhesive tape with the use of the modified silane-coated technique. The objective of this study was to compare section loss in slides using commercial adhesive tape, silane-coated microslides with the conventional technique or with the modified technique. Material and method: The TMA was constructed with hepatic tissue blocks embedded in paraffin, using a fixed base device, placing 32 cylinders of 2 mm in diameter in duplicate and 2.2 mm apart from each other. Fifteen 4-μm sections were placed on conventional silane-coated microslides at 4% (Group 1), 15 on silane-coated microslides with a modified technique (6% of silane and minimum use of acetone) (Group 2), and 15 on slides using commercial adhesive tape, according to the manufacturer's recommendations (Group 3). All microslides were processed by immunohistochemistry for cytokeratin 18, with antigen retrieval accomplished by incubation with citrate buffer pH 6.0 with microwave enhancement. Samples loss was quantified and expressed as: total (≥ 80%), almost complete (75% to 79%) or partial (50% to 74%). Results: The loss of sections was similar in all three groups (4.9 vs. 3.1 vs. 8.1, respectively) (analysis of variance [ANOVA], p = 0.3654). One slide usingcommercial adhesive tape showed artifactual ungluing of all sections and another one showed loss of 20 sampleson one side of the slide. None of the silane-coated microslides showed such artifact. Conclusions: Silane-coated microslides show adequate results, require less technical training and reduce the cost of TMA procedure, whatjustifies their use in research...

Procesamientos histológicos convencional y en microondas: comparación histológica, histoquímica e inmunohistoquímica / Conventional and microwave histological processing: a histological, histochemical and inmunohistochemical comparison

El horno microondas (HM) fue introducido en Anatomía Patológica en los años 80, siendo utilizado en varios procesos dentro del laboratorio. Se realizó la comparación de algunos parámetros histológicos, histoquímicos e inmunohistoquímicos en muestras de tejido normal, procesados paralelamente de manera convencional y en HM. Se seleccionaron muestras en duplicado de diversos tejidos; se procesaron convencionalmente (PC) en un procesador automático y en HM utilizando un protocolo estandarizado previamente. Se realizaron tinciones histológicas e histoquímicas (H-E, PAS y Azul Alcián) e inmunohistoquímicas (CD45, CD3, CD20 Citoqueratinas AE1/AE3, 5/18, 20, S-100, Receptores de Estrógeno y Progesterona). Las técnicas se escogieron según tipo de tejido. Se evaluó la calidad de los preparados (tinción nuclear, tinción citoplasmática y calidad general), la intensidad de las tinciones histológicas, histoquímicas e inmunohistoquímica de marcadores escogidos. El HM posibilitó la entrega de la lámina histológica en un tiempo mucho menor que el PC, disminuyendo el tiempo de procesamiento de 12 a 1 hora. Se observó una contracción mayor en las muestras procesadas en HM que en sus pares procesadas convencionalmente: tonsila palatina (54,8 por ciento v/s 46,6 por ciento), intestino grueso (33,3 por ciento v/s 22,2 por ciento), piel (24,1 por ciento v/s 13,7 por ciento) y mama (15,1 por ciento v/s 26,3 por ciento). La calidad del corte histológico fue mayor en las muestras PC, observando en los tejidos HM una tinción menos homogénea, con agrietamientos y desprendimientos más notorios, siendo igualmente apropiadas para el diagnóstico histológico. En cuanto a las tinciones histoquímicas, no se observaron cambios significativos. Se observó una mayor intensidad de tinción inmunohistoquímica de proteína S-100 y citoqueratina AE1/AE3 en piel. Se concluyó que el HM convencional puede ser utilizado en el procesamiento de biopsias sin inconvenientes para el diagnóstico...

The microwave oven (MO) was introduced in Pathology practice in the eighties and has been used in several processes within the laboratory. It was achieved a comparison between conventional and microwave processing. Histological, histochemical and inmunohistochemical parameters were observed. Samples of four kinds of tissues were taken in duplicate, and were processed on an automatic processor and into a microwave oven using a previous used protocol. H-E, PAS and Alcian Blue stains ware used in addition to inmunohistochemicals stains (CD45, CD3, CD20, Keratins AE1/AE3, 5/18 and 20, S-100 protein and estrogens and progesterone receptors). Techniques were chosen according to the tissue. The slides were evaluated for two blind professionals, whom considered: nuclear and cytoplasm stain, specificity and intensity of histochemical and inmunohistochemical stains. MO made possible a quick delivery of the slides, shorting the time from 12 to 1 hour. Shrinking was observed to be higher in MO process than conventional process: tonsils (54.8 percent v/s 46.6 percent), intestine (33.3 percent v/s 22.2 percent), skin (24.1 percent v/s 13.7 percent) y breast (15.1 percent v/s 26.3 percent). Qualification was found lightly higher on conventional processing. No changes were observed on histochemicals stains. Higher intensity of inmunostains was observed just in AE1/AE3 and S100 protein. It was concluded that MO can be used without inconveniences in the Pathology Laboratory, allowing a rapid diagnose of biopsy samples.

Cytomorphologic comparison between rehydrated air-dried and conventional wet-fixed pap smears.

OBJECTIVE: To compare the cytomorphologic quality of the cervical (Pap) smears between two fixation techniques, rehydration of air-dried smears (AD) versus wet fixation (WF). MATERIAL AND METHOD: Paired-cervical smears (AD and WF) from 172 women who underwent cervical cytology screening at Chiang Mai University Hospital between August 2004 and September 2004 were prospectively evaluated for the cytologic parameters and the staining qualities. RESULTS: The mean age of the 172 women was 41.7 years +/-2 SD 18.1), 27 women (15.7%) were postmenopausal. Absence ofred blood cells in the smear background was significantly more frequent in AD smears than in WF specimens (p = 0.0006). Air-drying artifact was more frequent in AD smears compared to those of WF (p = 0.036) but was of only mild degree in all cases. There was no significant difference between AD and WF smears in the cytoplasmic quality including distinctness of cell border (p = 0.30) and satisfactory staining (p = 0.054). For the nuclear morphology, there was no significant difference between both fixation techniques in the distinctness of nuclear border (p = 0.26) and chromatin crispness (p = 0.23) of the endocervical nuclei. In squamous nuclei, AD smears had higher frequency of indistinct nuclear border and hazy chromatin compared to WF smears (p = 0.003 each). However, these were observed in only mild degree and did not affect the cytologic interpretation. CONCLUSION: The quality of AD smears was slightly inferior to WF smears but was still satisfactory for cervical cytology. AD technique may be acceptable as an alternative to wet fixation in cytologic cervical cancer screening.

Extendidos delgados en la puncion aspiracion con aguja fina de tiroides: un estudio de 200 casos / Thin layer preparations in thyroid fine-needle aspiration: study of 200 cases

INTRODUCTION: Trying to increase the specificity and demonstrated sensibility of thyroid fine needle aspiration (TFNA) in the diagnosis of the thyroid nodule, and to improve the cytomorphologic diagnosis, we used the approach of realization of thin prep (TP), adapting the technique to the methods within our possibilities. MATERIALS AND METHODS: The sample corresponded to a prospective study of 200 cases of material of thyroid nodules, obtained by means of ultrasonography guide fine needle aspiration, over a 30 month period. The study was carried out by means of conventional smear (CS) and TP, cytologic preparations were stained with hematoxilin-eosine (H-E), Papanicolaou (PAP) and Tint 15. The analysis of these same samples was carried out according to sex, age, characteristics of the nodules and cytomorphologic features. RESULTS: The samples were satisfactory in 184 cases (92 per cent) and unsatisfactory in 16 cases (8 per cent). The prevalence in samples corresponded to the female sex (88.5 per cent). The pre-emptive diagnoses corresponded to: Multinodular Goiter: 41 (22.29 per cent)--Hashimoto's thyroiditis: 4 (2.17 per cent)--Follicular lesions I: 86(46.74 per cent), II: 34 (18.47 per cent), III: 6 (3.26 per cent)--Hurthle cell neoplasm: 4 (2.17 per cent)--Papillary carcinoma: 9 (4.90 per cent). The patient study was carried out according to the modified algorithm proposed by Gharib in 1997 and the Department of Cytology of the Karolinska Hospital of Stockholm (Sweden). The cytological diagnosis was compared to the final histological results. CONCLUSIONS: The present work has demonstrated that even with the available methods, we have been able to carry out TP that were acceptable for its evaluation and with the same limitations of those obtained by CS, with respect to preparations dried in direct air and in the cases of non-conclusive diagnoses. We obtained a sensibility of 94.44 per cent and a specificity of 98.79 per cent.

Role of potassium hydroxide mount in rapid diagnosis of fungal corneal ulcers.

The aim of the study was to establish the importance of KOH mount in rapid diagnosis of fungal corneal ulcers. Corneal scrapings were collected from 50 patients with corneal ulcers where clinical features suggested fungal aetiology and ulcers showed no signs of healing on initial treatment. Corneal scrapings were subjected to wet mount with 10% KOH, Gram's stain, and culture on SDA media. Wet mount showed fungal elements in 34 cases (68%) and were classified according to morphology. Gram's stain of corneal scrapings did not provide much help. Fungal culture was positive in 9 (26.4%) out of 34 cases. Specific antifungal treatment was instituted in all cases that showed positive wet mount without waiting for culture reports and on follow-up all these 34 patients showed improvement. Thus wet mount with 10% KOH can be relied upon as the singlemost important screening tool for rapid diagnosis of fungal corneal ulcer and treatment should be dispensed on its basis.