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.

Paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples / 浙江大学学报·医学版

OBJECTIVE@#To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples.@*METHODS@#p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis.@*RESULTS@#With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(=7.817 and 1.332, both >0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (=17.688 and 10.182, <0.05 or <0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick.@*CONCLUSIONS@#Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.

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)

Princípios do processamento histológico de rotina / Principles of histological processing

O presente estudo teve por objetivo realizar um breve histórico do desenvolvimento das técnicas de preparo de tecido para observação em microscopia óptica e descrever as fases do processamento histológico de rotina em laboratórios de anatomia patológica e compilar as principais variações em cada etapa, bem como observar as principais causas de falhas técnicas nestes procedimentos.

This study aimed to carry out a brief history of the development of tissue preparation techniques for observation with optical microscopy and describe the stages of histological processing in pathology laboratories and compile the main changes in each step and observe the main causes of technical flaws in these procedures.

Cytohistological correlation and discrepancy of conventional Papanicolaou smear test with corresponding histopathology: a retrospective study over a 5-year period

The accuracy of Papanicolaou [Pap] smear testing can be measured using the cytohistological correlation and discrepancy method. This study aimed to evaluate and compare the cytohistological correlation and discrepancy of the conventional Pap test with the corresponding histopathology and to compare the data with other similar studies. A retrospective study was performed at the pathology department of a referral hospital in Muscat, Oman, over a 5-year period. Of 6000 Pap smears, 162 had matching histopathology results [abnormal smear rate 2.7%] but 10 were unsatisfactory for histological diagnosis. Cytohistological correlation was seen in 96/152 [63.2%], while discrepancy was seen in 56/152 [36.8%]. False negatives and false positives were found for 7 and 49 cases respectively. The findings of this study confirm the role of conventional Pap testing as a screening test for the diagnosis of cervical lesion but not for management of patients. In comparison with other studies, we also report a low percentage of abnormal Pap smears

Correlación cito-histológica del tumor papilar de la glándula pineal. Presentación de un caso / Papillary pineal tumor: cyto-histological correlation. Case report

En la última revisión de la Organización Mundial de la Salud (OMS) en relación a los tumores del sistema nervioso central (SNC), se describieron nuevas entidades, como el Tumor Papilar de la Glándula Pineal. Esta lesión de rara aparición, se ha identificado en adultos jóvenes. El diagnóstico de estos tumores es complejo ya que depende de su ubicación, edad de aparición y el aspecto histológico; éste último tiene similitudes con otras lesiones como el ependimoma papilar o el papiloma/carcinoma de plexos coroides. Citológicamente presentan características claras que pueden ayudar al diagnóstico a través de la impronta en el estudio intraoperatorio; reconocer ciertos criterios con éste importante y sencillo método diagnóstico ha sido la motivación principal para el estudio de entidades poco frecuentes del SNC, además de corroborar el necesario trabajo de un equipo multidisciplinar.

In the latest revision of the central nervous system tumors (CNS) of the World Health Organization (WHO), new entities has been described, as papillary tumor of the pineal region. This rare lesion has been identified in young adults. The diagnosis of these tumors is complex, depends on the location, age of onset and histological appearance. Histological characteristics have similarities with other lesions such as papillary ependymoma, papiloma / choroid plexus carcinoma. Cytologically have clear characteristics that can aid in the diagnosis through the smears on the intraoperative study. Certain criteria for recognize this important and simple diagnostic method has been the main motivation for the study of CNS rare entities, as our case, in addition to corroborating the necessary work of a multidisciplinary team.

Performance effect of rabbit carotid artery treated with decellularization and photo-oxidation / 中南大学学报(医学版)

OBJECTIVE@#To analyze and discuss the feasibility of rabbit carotid artery treated with decellularization and photo-oxidation.@*METHODS@#Sixty vascular slices of rabbit carotid artery were divided into a fresh group, a cryopreservation group, a glutaraldehyde group, and a decellularization plus photo-oxidation group 15 in each group. To evaluate the physical properties of all the rabbit carotid arteries by testing heat-shrinking temperature, tensile stress and the max elongation of each group. Then by buliding subcutaneous embedding model in SD rats we evaluated the biological stability and the anti-calcification function property of the above rabbit carotid arteries, and the detection means included HE stain, atomic absorption spectrometry and Von-Kossa calcium salt stain.@*RESULTS@#The heat-shrinking temperature, tensile stress and the max elongation in the cryopreservation group were lower or shorter than those of the other groups and the difference had statistical significance (P<0.05). Although the heat-shrinking temperature and the tensile stress in the decellularization plus photo-oxidation group were lower or shorter than those in the glutaraldehyde group (P<0.05), the max elongation in the decellularization plus photo-oxidation group was much longer than that in the glutaraldehyde group (P<0.05). The rabbit carotid artery treated with decellularization plus photo-oxidation showed lower immunogenicity and better biological stability and better anti-calcification property compared with the other groups.@*CONCLUSION@#Decellularization associated with photo-oxidation is a suitable and novel protocol for small caliber artery allograft with a diameter of less than 6 mm which is unbreakable to mechanical properties and conducive to biological stability, which has a broad prospect.

Diafanización y tinción de hueso y cartílago en embriones y pequeños vertebrados / Diferenbtiated diafanization and staining of bones and cartilage in embryos and smal vertebrates

Este trabajo forma parte del Taler de Biología Aplicadaque se dicta en la Cátedra de Anatomía Comparada,Facultad de Ciencias Exactas Físicas y Naturales de laUniversidad Nacional de Córdoba. República Argentina. Objetivo: Aplicar Técnicas macroscópicas convencionales y nuevas estrategias en la preparación y exhibición de embriones y pequeños vertebrados diafanizados.Materiales y métodos: Se utilzaron un total de seisespecímenes representativos de distintos vertebrados:un juvenil del pez chanchita (Australoherus facetus), dosadultos: un anfibio, la ranita lorona (Physalaemusbilngonigerus) y un reptil, vulgarmente conocido comolagarto gusano (Amphisbaenidae) y tres embriones: tortuga terestre (Chelonidis chilensis), paloma torcaza(Zenaida auriculata) y comadreja roja (Lutreolinacrasicaudata). El material biológico fue fijado en formol4%, conservado en alcohol 70% y luego procesado deacuerdo al siguiente protocolo: 1) Identifcación del espécimen, 2) Morfometría, 3) Extración del tegumento,evisceración, remoción de encéfalo y enucleación delos ojos. 4) Lavado, 5) Blanqueamiento en Peróxido deHidrógeno, 6) Inmersión en una solución de Azul de Alcianpara teñir cartílago y postfijación en alcohol 10%, 7)Inmersión en una solución de KHO, a partir del 3% por24 hs, 8) El tejido óseo se coloreó con el agregado, a lasolución de KHO, de 8 gotas de solución acuosa dealizarina roja en 10 ml, controlando la transparencia yel color. 9) Transferencia de las muestras a solucionesde glicerina al 25, 50 y 75% por 24 hs y luego conservación de las mismas en glicerina anhidra pura en un recipiente etiquetado. Resultados y conclusiones: Con el usode esta técnica se obtuvieron muy buenos resultados enla diafanización de los tejidos, coloración de hueso ycartílago y en la integridad de los ejemplares. Actualmente, este material biológico se conserva en la Cátedra de Anatomía Comparada.

This research work is part of the Aplied Biology Workshopheld at he Cátedra de Anatomía Comparada de la Facultadde Ciencias Exactas, Físicas y Naturales de laUniversidad Nacional de Córdoba. R. Argentina.Objective: It was caried out with the purpose of aplyingconventional macroscopic techniques and newstrategies in the preparation and exhibiton of diafanizedembryos and smal vertebrates. Materials and Methods:Six specimens representative of diferent vertebrates wereused: a young “Chemeleon Cichlid” (Australoherusfacetus); two adults: an amphibian (Physalaemusbilngonigerus), “crying frog” a reptile, (Amphisbaenidae),commonly known as worm lizard and thre embryos: aland turtle, (Chelonidis chilensis), an Eared Dove (Zenaidaauriculata), and a lutrine or thick-tailed oposum(Lutreolina crasicaudata). The biological material wasfixed in 4% formalin, preserved in 70% alcohol andprocesed acording to the folowing procedure:1)Specimen identifcation. 2) Morphometry. 3) Tegumentremoval, evisceration, encephalon removal andenucleation of the eyes. 4) Washing. 5) Whitening withhydrogen peroxide. 6) Immersion in a solution of AlcianBlue staining to reveal cartilage, and post fixationtreatment in alcohol 10%, 7) Immersion in a solution ofKOH (potasium hydroxide), starting at 3% for 24 hours.8) In order to colour the bone tisue, 8 drops of alizarinred aqueous solution were aded in 10 ml to theimmersion solution of KOH while transparency and colorwere controled. 9) The samples were transfered toglycerin solutions 25, 50 and 75% for 24 hours and thenpreserved in pure anhydrous glycerin in labeledcontainers. Results and conclusions: With using of thistechnique were obtained very god results in thediafanization of tisue, colouring of bone and cartilageand integrity of the specimens. At present, this biologicalmaterial is kept at he Cátedra de Anatomía Comparada.

An improved protocol of preparing bone marrow cells for fluorescence in situ hybridization / 中国实验血液学杂志

This study was aimed to establish a smear protocol for preparing bone marrow cells and investigate its effect on fluorescence in situ hybridization (FISH) signal. Probe DNA (C-myc, MDM2, STK6) was labeled with Spectrum Green, PromoFluor-555 and PromoFluor-415 by nick translation. Five bone marrow samples were tested by two methods separately. Traditional method: after removing the erythrocytes by hypoosmotic solution, the bone marrow cells were fixed in methanol/acetic acid (3:1). Improved method: erythrocytes were removed using density gradient centrifugation and fixed in methanol. The samples were then fixed again in 2 formaldehyde for 5 min. The FISH signal was assessed by comparing the relative signal intensity of each fluorophore with the autofluorescence background. The results indicated that improved method greatly increased the ratio of fluorescence signal intensity in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel (traditional method: 4.3 ± 0.19, 3.52 ± 0.04, 3.07 ± 0.08; improved method: 9.89 ± 0.41, 7.55 ± 0.5, 5.67 ± 0.18, n = 5, P < 0.01) respectively. The signal intensity increased 2.32, 2.14 and 1.85-fold in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel respectively. In addition, the improved method decreased the split signals [traditional method: (15.8 ± 1.74), (20.42 ± 2.88), (23.2 ± 3.02); improved method: (8.6 ± 1.2), (12.28 ± 1.33), (12.6 ± 2.56), n = 5, P < 0.05]. It is concluded that the improved optimal procedure which facilitates FISH intensity on bone marrow cells is developed, showing potential for wide application in the diagnosis of hematologic diseases.

A clinical and molecular cytogenetic study of Filipino patients with williams syndrome

Objective. To study the clinical spectrum of Filipino patients with Williams Syndrome and to confirm the gene deletion by FISH analysis. Methods. From June 2005 to September 2008, patients who were seen at the Genetics clinic of the UP-PGH and who met the clinical criteria for Williams Syndrome were analyzed for the 7q11.23 deletion through karyotyping and FISH studies. A detailed history and a thorough dysmorphologic examination were performed. Relevant investigations included two-dimensional echocardiography, renal ultrasonography, ophthalmologic examination, developmental assessment and serum calcium determination. Result. Eight patients were included in the study. The mean age at first diagnosis was 8.5 years. All cases were sporadic. The chromosomal analysis was normal for all patients and in the FISH analysis, a 7q11.23 deletion was detected in 100% of cases. Distinctive facial features, cardiac abnormalities and developmental delay were present in all patients. The typical behavior of overfriendliness was observed in the majority of cases. Hypercalcemia was documented in only one case and no renal anomalies were detected. Conclusion. The craniofacial features were similar among patients but there is a broad spectrum of severity of clinical features in cardiovascular abnormalities, personality, behavior traits and mental capacity.

Influence of amplicon size on the polymerase chain reaction of Parvovirus B19 genome in formalin-fixed specimens / Influência do tamanho do amplicon na reação em cadeia da polimerase na detecção do genoma do PB19 em amostras fixadas em formalina

The polymerase chain reaction (PCR) has provided diagnosis of archival material, but some fixation methods such as formalin damage DNA and, subsequently, affect PCR analysis, particularly paraffin-embedded tissues. PCR is known due to its high specificity and sensitivity, although some difficulties arise when formalinfixed and paraffin-embedded tissue is used. Not only does this occur due to protein cross-linking, which increases with longer fixation time, but it also happens due to the direct damage that formalin causes in the DNA itself. PCR was used to analyze placenta and fetal organs from 34 samples with suspected Parvovirus B19 infection. It was not possible to amplify Parvovirus B19 DNA using nested-PCR, probably due to the size of the amplicon generated with the first set of primers. We approached this problem by using only the second set of primers. Two out of 34 tissue samples (5,9 percent) were positive by PCR. However, PCR performed on corresponding fetal organs was negative in one of the two. We also observed a negative relation between the thickness of the tissue fragment and the positivity of the samples. In conclusion, although PCR is highly specific and sensitive in fresh or ideally fixed material, a careful standardization of PCR assays is necessary when using formalin fixed paraffin-embedded tissues by applying primers that require smaller DNA fragments for amplification.

A reação em cadeia da polimerase (PCR) tem fornecido diagnóstico de material de arquivo, mas alguns métodos de fixação, tais como formalina, provocam danos ao DNA e subsequentemente afetam sua análise, particularmente tecidos embebidos em parafina. A PCR é conhecida pela sua alta especificidade e sensibilidade, embora algumas dificuldades ocorram quando o material utilizado foi fixado em formalina e embebido em parafina. Isso não se deve somente pela formação de cross-linkings com proteínas, a qual aumenta com o maior tempo de fixação, mas também pelo dano direto que a formalina causa no DNA. PCR foi usada para analisar placenta e órgão fetais de 34 amostras com suspeita de infecção pelo Parvovírus B19 (PB19). Não foi possível amplificar o DNA do PB19 usando nested-PCR, provavelmente devido ao tamanho do amplicon gerado com o primeiro passo dos primers. Adequamos o problema utilizando somente o segundo par de primers e pudemos observar que das 34 amostras, duas eram positivas para PCR (5,9 por cento). Entretanto, a PCR dos órgãos fetais foi negativa em um de dois casos. Observamos também uma relação negativa entre a espessura do corte dos materiais com a positividade das amostras. Em conclusão, embora a PCR seja altamente específica e sensível em amostras a fresco ou idealmente fixadas, uma padronização cuidadosa para análise com a PCR é necessária quando se utiliza tecidos fixados em formalina e embebidos em parafina utilizando primers que requerem menor fragmento de DNA para a amplificação.

Estudo retrospectivo dos diagnósticos cito-histopatológicos nas atipias de significado indeterminado / Retrospective study on cyto-histological diagnoses in the atypical squamous cells of undertermined signifance

No presente estudo foi avaliada a história de realização dos exames citológicos (EC) prévios e posteriores com diagnóstico citológico de células escamosas atípicas (ASC) e sua correlação com os resultados de exames de biópsias. Para tanto, foi realizado o estudo retrospectivo, de 60 casos de EC cérvico-vaginais com diagnóstico de ASC e biópsias correlatas, os exames prévios e posteriores de ASC e a periodicidade de exames realizados. Das 60 análises selecionadas, 57 apresentaram diagnóstico citológico anterior de ASC-US (possivelmente não neoplásico), 30 (52,6%) foram negativos no exame de biópsia, 16 (28,1%) de NIC 1, 11 (19,3%) com diagnósticos discrepantes acima de 2 graus; 3 (100%) casos de ASC-H (não se pode afastar lesão de alto grau) foram NIC 1 na biópsia. Dentre os 60 casos, 44 (73,3%) que tiveram exames anteriores, 14 (23,3%) eram positivos; 53 (88,3%) que fizeram EC posteriores à biópsia, 10 (16,7%) eram positivos. Quanto à periodicidade dos exames, 26 (43,3%) realizaram o exame com intervalo de até 1 ano, 29 (48,3%) de 1 a 3 anos, 1 (1,7%) em intervalo maior que 3 anos. De acordo com os achados observados neste estudo, os diagnósticos citológicos de ASC, quando comparado com os exames da biópsia, estavam mais frequentemente associados às alterações morfológicas reacionais. Estes dados mostram o valioso papel da correlação cito- histológica, uma vez que a execução apenas do seguimento citológico pode ser insuficiente para obter o diagnóstico morfológico. É salientada, ainda, a necessidade de efetuar o exame de acompanhamento de seis em seis meses, para realizar o controle evolutivo dessas atipias, bem como para auxiliar na conduta terapêutica das pacientes.

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.

The value of thin-layer cytology in intraoperative fine-needle aspiration biopsy of pancreatic cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The purpose of this study was to explore the value of thin-layer cytology (TLC) in intraoperative fine needle aspiration cytology diagnosis of pancreatic cancer.</p><p><b>METHODS</b>Results of cytological examination with thin-layer smears were compared with that with conventional smears in intraoperative fine-needle aspiration (FNA) biopsy.</p><p><b>RESULTS</b>Totally 271 fine needle aspiration biopsies were performed, among them, 70 were examined with thin-layer smears, showing unsatisfactory smear in 5 cases (7.1%); 201 were examined with conventional smears (CS), showing unsatisfactory smear in 9 cases (4.5%). No significant difference in the unsatisfactory smears was observed between those two groups. The positive rate of diagnosis with CS smears was 60.0% (42/70) and that of TLC was 81.6% (164/201), with a significant difference (P < 0.01). The sensitivity of CS and TLC was 68.9% and 87.7%, respectively (P < 0.01). The sensitivity of both FNA and frozen section diagnosis in 20 cases was 90.0%, respectively, but that of FNA combined with frozen section diagnosis was 95.0%. 9 cancer cases diagnosed by pathology were initially negative by cytology, but adenocarcinoma cells were found in 7 cases of them by the second time cytology examination.</p><p><b>CONCLUSION</b>The positive rate is high in TLC smears, and unsatisfactory rate is low. TLC smears are one of the best methods in intraoperative confirmation of pancreatic cancer. The use of FNA smears combined with frozen section biopsy can further improve the sensitivity of diagnosis.</p>

[Histopathologic evaluation of the pulp after direct pulp capping with calcium mineral trioxide aggregate in primary teeth]

Mineral trioxide aggregate [MTA] is a relatively new material. It is biocompatible and has sealing ability. The purpose of this study was to evaluate the histologic pulp responses of calcium hydroxide and MTA placed directly on exposed pulp tissues. Twenty teeth already scheduled for extraction for orthodontic reasons were selected for this experimental study. These teeth were divided into 2 groups and treated with direct pulp capping. Calcium hydroxide was used for 10 teeth and MTA for 10 teeth. After 60 days the teeth were extracted and prepared for histological evaluation. Finally the data were analyzed with Mann-Whitney test. All teeth treated with Calcium hydroxide showed inflammation. Internal resorption was seen in 6 teeth and abscess in 5 teeth and calcified bridge in 6 teeth and reparative in 2 teeth. Inflammation was seen in 2 mineral trioxide aggregate samples and calcified bridge and reparative dentin in 7 teeth. Internal resorption and abscess were not seen in mineral trioxide aggregate group. Mineral trioxide aggregate appeared to be superior to Calcium hydroxide as a pulp capping agent in primary teeth

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.

[Rapid preparation of tissues sections by microowave and comparison with routine method]

Microwave energy is produced from electricity energy via a magnetron which is discovered about 60 years ago and according to its several capabilities, is used in most industries. One of them is laboratory utility, especially in pathology laboratory, which was introduced in 1980 decade by Boon et.al. Microwave doesn't use in IRAN because ofexpensivity. Histoprocessing in pathology laboratory takes time about 24 hours and diagnosis is delayed one working day. For reducing time in diagnosis and decreasing hospital and insurance costs, and introducing an applicable way, the aim of this study is microwave oven usage in optimal tissue processing. ln this study, each level of processing was done separately and finally all the levels were combined then results were verified. According to details of tissue processing total time for all levels was 70 minutes. Distinction between each paithem was impossible and in many of them, microwave processing slides were better. Microwave processing could be used simply by pathology laboratories for reducing time ofhistoprocessing if correct calibration and setup is done

Watershed-based segmentation of histiocytic images / 生物医学工程学杂志

The task of segmenting histiocyte is a crucial step in the analysis of histiocytic images which is an important application of computer vision to histopathology. The algorithm presented in this article was composed of two steps: (1) the morph-based preprocessing; (2) the ameliorated watershed method. In the first step, the difference between histiocytes was magnified in order to increase the visibility from the view of the computer vision, and then the ameliorated terrace-flooding-simulated watershed method was used to achieve the segmentation of histiocytic images in the second step. To test the performance of the algorithm, different samples of visual quality were tested and the result figures proved successful.