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[PMID]:28453669
[Au] Autor:Hanslovsky P; Bogovic JA; Saalfeld S
[Ad] Endereço:HHMI Janelia Research Campus, Ashburn, VA 20147, USA.
[Ti] Título:Image-based correction of continuous and discontinuous non-planar axial distortion in serial section microscopy.
[So] Source:Bioinformatics;33(9):1379-1386, 2017 05 01.
[Is] ISSN:1367-4811
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Motivation: Serial section microscopy is an established method for detailed anatomy reconstruction of biological specimen. During the last decade, high resolution electron microscopy (EM) of serial sections has become the de-facto standard for reconstruction of neural connectivity at ever increasing scales (EM connectomics). In serial section microscopy, the axial dimension of the volume is sampled by physically removing thin sections from the embedded specimen and subsequently imaging either the block-face or the section series. This process has limited precision leading to inhomogeneous non-planar sampling of the axial dimension of the volume which, in turn, results in distorted image volumes. This includes that section series may be collected and imaged in unknown order. Results: We developed methods to identify and correct these distortions through image-based signal analysis without any additional physical apparatus or measurements. We demonstrate the efficacy of our methods in proof of principle experiments and application to real world problems. Availability and Implementation: We made our work available as libraries for the ImageJ distribution Fiji and for deployment in a high performance parallel computing environment. Our sources are open and available at http://github.com/saalfeldlab/section-sort, http://github.com/saalfeldlab/z-spacing and http://github.com/saalfeldlab/z-spacing-spark. Contact: saalfelds@janelia.hhmi.org. Supplementary information: Supplementary data are available at Bioinformatics online.
[Mh] Termos MeSH primário: Algoritmos
Metodologias Computacionais
Interpretação de Imagem Assistida por Computador/métodos
Microscopia Eletrônica/métodos
[Mh] Termos MeSH secundário: Animais
Sistema Nervoso Central/anatomia & histologia
Drosophila melanogaster/anatomia & histologia
Microtomia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170429
[St] Status:MEDLINE
[do] DOI:10.1093/bioinformatics/btw794


  2 / 3913 MEDLINE  
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[PMID]:29254315
[Au] Autor:Akhtar T; Sheikh N; Shan T; Ghazanfar R
[Ad] Endereço:Cell and Molecular Biology Lab, Department of Zoology, University of the Punjab, Q-A Campus, Lahore, Pakistan.
[Ti] Título:Tacrolimus induced nephrotoxicity and pulmonary toxicity in Wistar rats.
[So] Source:J Biol Regul Homeost Agents;31(4):1061-1066, 2017 Oct-Dec.
[Is] ISSN:0393-974X
[Cp] País de publicação:Italy
[La] Idioma:eng
[Ab] Resumo:Transplantation has evolved into an accepted treatment for end-stage organ failure. The major limitation for solid organ transplantation is organ rejection, which is an adaptive immune response caused by the activation of T-cells. Immunosuppressant drugs are used to overcome this problem. Tacrolimus is a powerful immunosuppressive drug which is used to minimize the risk of organ rejection. The present study was designed to find the toxic effects of tacrolimus on lungs and kidneys. Wistar rats were divided into 4 experimental groups and one control group (n=9). Each rat of the experimental group was orally given the aqueous suspension of tacrolimus powder (3mg/ml) and dissected after 6, 12, 24 and 48 hours of tacrolimus suspension dose. Lungs and kidneys were excised and processed for histopathological and histochemical alterations. Kidney tissues presented signs of toxic potential on tissue architecture such as increased interstitial spaces, necrosis, especially acute tubular necrosis, glomerular shrinkage, dilated blood vessels and enlargement of Bowman’s capsule. Lung sections also confirmed the toxic potential, characterized by bronchiolar wall thickening, alveolar cells necrosis, collapsing of alveolar spaces and interstitial round cell infiltrate. Results of Prussian blue iron staining showed no iron deposition in kidney architecture while in lung sections, iron accumulation was evident. Taken together from these observations we can conclude that tacrolimus may induce toxicity to a certain extent with structural distortion of the kidneys and lungs.
[Mh] Termos MeSH primário: Imunossupressores/toxicidade
Rim/efeitos dos fármacos
Pulmão/efeitos dos fármacos
Tacrolimo/toxicidade
[Mh] Termos MeSH secundário: Animais
Ferrocianetos
Histocitoquímica
Rim/patologia
Rim/ultraestrutura
Pulmão/patologia
Pulmão/ultraestrutura
Microtomia
Ratos
Ratos Wistar
Inclusão do Tecido
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Ferrocyanides); 0 (Immunosuppressive Agents); TLE294X33A (ferric ferrocyanide); WM0HAQ4WNM (Tacrolimus)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180222
[Lr] Data última revisão:
180222
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171220
[St] Status:MEDLINE


  3 / 3913 MEDLINE  
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[PMID]:29189769
[Au] Autor:Watson SA; Scigliano M; Bardi I; Ascione R; Terracciano CM; Perbellini F
[Ad] Endereço:Division of Cardiovascular Sciences, Myocardial Function, National Heart and Lung Institute, Imperial College London, London, UK.
[Ti] Título:Preparation of viable adult ventricular myocardial slices from large and small mammals.
[So] Source:Nat Protoc;12(12):2623-2639, 2017 Dec.
[Is] ISSN:1750-2799
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:This protocol describes the preparation of highly viable adult ventricular myocardial slices from the hearts of small and large mammals, including rodents, pigs, dogs and humans. Adult ventricular myocardial slices are 100- to 400-µm-thick slices of living myocardium that retain the native multicellularity, architecture and physiology of the heart. This protocol provides a list of the equipment and reagents required alongside a detailed description of the methodology for heart explantation, tissue preparation, slicing with a vibratome and handling of myocardial slices. Supplementary videos are included to visually demonstrate these steps. A number of critical steps are addressed that must be followed in order to prepare highly viable myocardial slices. These include identification of myocardial fiber direction and fiber alignment within the tissue block, careful temperature control, use of an excitation-contraction uncoupler, optimal vibratome settings and correct handling of myocardial slices. Many aspects of cardiac structure and function can be studied using myocardial slices in vitro. Typical results obtained with hearts from a small mammal (rat) and a large mammal (human) with heart failure are shown, demonstrating myocardial slice viability, maximum contractility, Ca handling and structure. This protocol can be completed in ∼4 h.
[Mh] Termos MeSH primário: Ventrículos do Coração/citologia
Microtomia/métodos
Miocárdio/citologia
[Mh] Termos MeSH secundário: Adulto
Animais
Soluções Cardioplégicas/química
Dissecação/métodos
Cães
Desenho de Equipamento
Seres Humanos
Indicadores e Reagentes
Camundongos
Microtomia/instrumentação
Ratos
Preservação de Tecido/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cardioplegic Solutions); 0 (Indicators and Reagents)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171219
[Lr] Data última revisão:
171219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171201
[St] Status:MEDLINE
[do] DOI:10.1038/nprot.2017.139


  4 / 3913 MEDLINE  
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[PMID]:29040322
[Au] Autor:Meleis AM; Mahtabfar A; Danish S; Foty RA
[Ad] Endereço:Department of Surgery, Rutgers Robert Wood Johnson Medical School, New Brunswick, New Jersey, United States of America.
[Ti] Título:Dexamethasone-mediated inhibition of Glioblastoma neurosphere dispersal in an ex vivo organotypic neural assay.
[So] Source:PLoS One;12(10):e0186483, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Glioblastoma is highly aggressive. Early dispersal of the primary tumor renders localized therapy ineffective. Recurrence always occurs and leads to patient death. Prior studies have shown that dispersal of Glioblastoma can be significantly reduced by Dexamethasone (Dex), a drug currently used to control brain tumor related edema. However, due to high doses and significant side effects, treatment is tapered and discontinued as soon as edema has resolved. Prior analyses of the dispersal inhibitory effects of Dex were performed on tissue culture plastic, or polystyrene filters seeded with normal human astrocytes, conditions which inherently differ from the parenchymal architecture of neuronal tissue. The aim of this study was to utilize an ex-vivo model to examine Dex-mediated inhibition of tumor cell migration from low-passage, human Glioblastoma neurospheres on multiple substrates including mouse retina, and slices of mouse, pig, and human brain. We also determined the lowest possible Dex dose that can inhibit dispersal. Analysis by Two-Factor ANOVA shows that for GBM-2 and GBM-3, Dex treatment significantly reduces dispersal on all tissue types. However, the magnitude of the effect appears to be tissue-type specific. Moreover, there does not appear to be a difference in Dex-mediated inhibition of dispersal between mouse retina, mouse brain and human brain. To estimate the lowest possible dose at which Dex can inhibit dispersal, LogEC50 values were compared by Extra Sum-of-Squares F-test. We show that it is possible to achieve 50% reduction in dispersal with Dex doses ranging from 3.8 x10-8M to 8.0x10-9M for GBM-2, and 4.3x10-8M to 1.8x10-9M for GBM-3, on mouse retina and brain slices, respectively. These doses are 3-30-fold lower than those used to control edema. This study extends our previous in vitro data and identifies the mouse retina as a potential substrate for in vivo studies of GBM dispersal.
[Mh] Termos MeSH primário: Antineoplásicos Hormonais/farmacologia
Movimento Celular/efeitos dos fármacos
Dexametasona/farmacologia
Neuroglia/efeitos dos fármacos
Esferoides Celulares/efeitos dos fármacos
[Mh] Termos MeSH secundário: Animais
Encéfalo/citologia
Relação Dose-Resposta a Droga
Células Alimentadoras/citologia
Seres Humanos
Camundongos
Microtomia
Neuroglia/patologia
Retina/citologia
Esferoides Celulares/patologia
Suínos
Células Tumorais Cultivadas
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antineoplastic Agents, Hormonal); 7S5I7G3JQL (Dexamethasone)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171031
[Lr] Data última revisão:
171031
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171018
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0186483


  5 / 3913 MEDLINE  
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[PMID]:28739252
[Au] Autor:Szulczyk B; Nurowska E
[Ad] Endereço:Department of Drug Technology and Pharmaceutical Biotechnology, The Medical University of Warsaw, Banacha 1, 02-097, Poland. Electronic address: bartlomiej.szulczyk@wum.edu.pl.
[Ti] Título:Valproic acid inhibits TTX-resistant sodium currents in prefrontal cortex pyramidal neurons.
[So] Source:Biochem Biophys Res Commun;491(2):291-295, 2017 Sep 16.
[Is] ISSN:1090-2104
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Valproic acid is frequently prescribed and used to treat epilepsy, bipolar disorder and other conditions. However, the mechanism of action of valproic acid has not been fully elucidated. The aim of this study was to assess the influence of valproic acid (200 µM) on TTX-resistant sodium currents in mPFC pyramidal neurons. Valproic acid inhibited the maximal amplitude and did not change the activation parameters of TTX-resistant sodium currents. Moreover, valproic acid (2 µM and 200 µM) shifted the TTX-resistant sodium channel inactivation curve towards hyperpolarisation. In the presence of valproic acid, TTX-resistant sodium currents recovered from inactivation more slowly. Valproic acid did not influence the use-dependent blockade of TTX-resistant sodium currents. This study suggests that a potential new mechanism of the antiepileptic action of valproic acid is, among others, inhibition of TTX-resistant sodium currents.
[Mh] Termos MeSH primário: Anticonvulsivantes/farmacologia
Células Piramidais/efeitos dos fármacos
Bloqueadores dos Canais de Sódio/farmacologia
Canais de Sódio/metabolismo
Tetrodotoxina/farmacologia
Ácido Valproico/farmacologia
[Mh] Termos MeSH secundário: Potenciais de Ação/efeitos dos fármacos
Potenciais de Ação/fisiologia
Animais
Técnicas de Cultura de Células
Microtomia
Técnicas de Patch-Clamp
Córtex Pré-Frontal/citologia
Córtex Pré-Frontal/efeitos dos fármacos
Córtex Pré-Frontal/metabolismo
Cultura Primária de Células
Células Piramidais/citologia
Células Piramidais/metabolismo
Ratos
Sódio/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anticonvulsants); 0 (Sodium Channel Blockers); 0 (Sodium Channels); 4368-28-9 (Tetrodotoxin); 614OI1Z5WI (Valproic Acid); 9NEZ333N27 (Sodium)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170824
[Lr] Data última revisão:
170824
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170726
[St] Status:MEDLINE


  6 / 3913 MEDLINE  
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[PMID]:28596154
[Au] Autor:Trout AT; Batie MR; Gupta A; Sheridan RM; Tiao GM; Towbin AJ
[Ad] Endereço:Department of Radiology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
[Ti] Título:3D printed pathological sectioning boxes to facilitate radiological-pathological correlation in hepatectomy cases.
[So] Source:J Clin Pathol;70(11):984-987, 2017 Nov.
[Is] ISSN:1472-4146
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Radiogenomics promises to identify tumour imaging features indicative of genomic or proteomic aberrations that can be therapeutically targeted allowing precision personalised therapy. An accurate radiological-pathological correlation is critical to the process of radiogenomic characterisation of tumours. An accurate correlation, however, is difficult to achieve with current pathological sectioning techniques which result in sectioning in non-standard planes. The purpose of this work is to present a technique to standardise hepatic sectioning to facilitateradiological-pathological correlation. We describe a process in which three-dimensional (3D)-printed specimen boxes based on preoperative cross-sectional imaging (CT and MRI) can be used to facilitate pathological sectioning in standard planes immediately on hepatic resection enabling improved tumour mapping. We have applied this process in 13 patients undergoing hepatectomy and have observed close correlation between imaging and gross pathology in patients with both unifocal and multifocal tumours.
[Mh] Termos MeSH primário: Hepatectomia
Neoplasias Hepáticas/diagnóstico por imagem
Neoplasias Hepáticas/patologia
Fígado/diagnóstico por imagem
Fígado/patologia
Imagem por Ressonância Magnética/instrumentação
Microtomia/instrumentação
Impressão Tridimensional
Tomografia Computadorizada por Raios X/instrumentação
[Mh] Termos MeSH secundário: Adolescente
Projeto Auxiliado por Computador
Desenho de Equipamento
Feminino
Seres Humanos
Lactente
Fígado/cirurgia
Neoplasias Hepáticas/cirurgia
Masculino
Valor Preditivo dos Testes
Interpretação de Imagem Radiográfica Assistida por Computador
[Pt] Tipo de publicação:CASE REPORTS; JOURNAL ARTICLE
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170610
[St] Status:MEDLINE
[do] DOI:10.1136/jclinpath-2016-204293


  7 / 3913 MEDLINE  
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[PMID]:28410135
[Au] Autor:Chu W; Zhao H; Li J; Li C
[Ad] Endereço:Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, State Key Lab for Molecular Science of Special AnimalsChangchun, Beijing, China.
[Ti] Título:Custom-built tools for the study of deer antler biology.
[So] Source:Front Biosci (Landmark Ed);22:1622-1633, 2017 Jun 01.
[Is] ISSN:1093-4715
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Deer antlers can be developed into multiple novel models to study growth and development of tissues for biomedical research. To facilitate this process, we have invented and further refined five custom-built tools through three decades of antler research. These are: 1. Pedicle growth detector to pinpoint the timing when pedicle growth is initiated, thus stimuli for pedicle and first antler formation can be investigated and identified. 2. Thin periosteum slice cutter to thinly slice (0.2mm or 0.7 mm thick) a whole piece of antlerogenic periosteum (AP) or pedicle periosteum (PP), which facilitates gene delivery into cells resident in these tissues, thus making transgenic antlers possible. 3. The porous periosteum multi-needle punch to effectively loosen the dense AP or PP tissue. This allows most cells of the periosteum to come into direct contact with treating solutions, thus making artificial manipulation of antler development possible. 4. The intra-dermal pocket maker to cut the thin dermal tissue (less than 2 mm in thickness) of a male deer calf horizontally into two layers to make an intra-dermal pocket. This allows loading of AP tissue intra-dermally to test the theory of "antler stem cell niche" . 5. The sterile periosteum sampling system to allow aseptic collection of the AP, PP or the antler growth centre tissue on farm, thus allowing antler generation, regeneration or rapid growth to be investigated . Overall, we believe the application of contemporary cellular and molecular biological techniques coupled with these custom-built tools would greatly promote the establishment of this unique and novel model for the benefits of biomedical research, and hence human health.
[Mh] Termos MeSH primário: Chifres de Veado/fisiologia
Cervos/fisiologia
Periósteo/fisiologia
Regeneração
Manejo de Espécimes/instrumentação
Manejo de Espécimes/métodos
[Mh] Termos MeSH secundário: Animais
Chifres de Veado/anatomia & histologia
Cervos/anatomia & histologia
Dissecação/instrumentação
Dissecação/métodos
Masculino
Microtomia/instrumentação
Microtomia/métodos
Modelos Animais
Periósteo/anatomia & histologia
Medicina Regenerativa/instrumentação
Medicina Regenerativa/métodos
Medicina Regenerativa/tendências
Pesquisa/tendências
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170831
[Lr] Data última revisão:
170831
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170415
[St] Status:MEDLINE


  8 / 3913 MEDLINE  
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[PMID]:28375970
[Au] Autor:Godsey T; Jacobson R; Gloster H
[Ad] Endereço:Department of Dermatology, University of Cincinnati, Cincinnati, Ohio.
[Ti] Título:Large Elliptical Specimens and the Single Section Method.
[So] Source:Dermatol Surg;43(9):1189-1191, 2017 09.
[Is] ISSN:1524-4725
[Cp] País de publicação:United States
[La] Idioma:eng
[Mh] Termos MeSH primário: Microtomia/métodos
Cirurgia de Mohs/métodos
Neoplasias Cutâneas/cirurgia
[Mh] Termos MeSH secundário: Seres Humanos
Margens de Excisão
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171012
[Lr] Data última revisão:
171012
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170405
[St] Status:MEDLINE
[do] DOI:10.1097/DSS.0000000000001071


  9 / 3913 MEDLINE  
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[PMID]:28274266
[Au] Autor:Niehof M; Hildebrandt T; Danov O; Arndt K; Koschmann J; Dahlmann F; Hansen T; Sewald K
[Ad] Endereço:Division of Preclinical Pharmacology and In Vitro Toxicology, Fraunhofer Institute for Toxicology and Experimental Medicine ITEM, German Center for Lung Research (DZL), Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Excellence Cluster REBIRTH, Nikolai-Fuchs-Str. 1, 3
[Ti] Título:RNA isolation from precision-cut lung slices (PCLS) from different species.
[So] Source:BMC Res Notes;10(1):121, 2017 Mar 09.
[Is] ISSN:1756-0500
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Functional 3D organ models such as precision-cut lung slices (PCLS) have recently captured the attention of biomedical research. To enable wider implementation in research and development, these new biologically relevant organ models are being constantly refined. A very important issue is to improve the preparation of high-quality RNA (ribonucleic acid) from PCLS for drug discovery and development of new therapies. Gene expression analysis at different levels is used as an important experimental readout. Genome-wide analysis using microarrays is mostly applied for biomarker selection in disease models or in comprehensive toxicological studies. Specific biomarker testing by reverse transcriptase quantitative polymerase chain reaction (RTqPCR) is often used in efficacy studies. Both applications require high-quality RNA as starting material for the generation of reliable data. Additionally, a small number of slices should be sufficient for satisfactory RNA isolation to allow as many experimental conditions as possible to be covered with a given tissue sample. Unfortunately, the vast amount of agarose in PCLS impedes RNA extraction according to the standard procedures. RESULTS: We established an optimized protocol for RNA isolation from PCLS from humans, rats, mice, marmosets, and rhesus macaques based on the separation of lysis and precipitation steps and a magnetic-bead cleanup procedure. The resulting RNA is of high purity and possesses a high degree of integrity. There are no contaminations affecting RTqPCR efficiency or any enzymatic step in sample preparation for microarray analysis. CONCLUSIONS: In summary, we isolated RNA from PCLS from different species that is well suited for RTqPCR and for microarray analysis as downstream applications.
[Mh] Termos MeSH primário: Pulmão/química
Microtomia/métodos
Análise de Sequência com Séries de Oligonucleotídeos/métodos
RNA/isolamento & purificação
Transcriptoma
[Mh] Termos MeSH secundário: Idoso
Animais
Callithrix
Feminino
Seres Humanos
Pulmão/cirurgia
Macaca mulatta
Imãs
Masculino
Camundongos
Camundongos Endogâmicos BALB C
Análise em Microsséries
Microtomia/instrumentação
Meia-Idade
Análise de Sequência com Séries de Oligonucleotídeos/estatística & dados numéricos
RNA/genética
Ratos
Ratos Wistar
Reação em Cadeia da Polimerase Via Transcriptase Reversa
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
63231-63-0 (RNA)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170317
[Lr] Data última revisão:
170317
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170310
[St] Status:MEDLINE
[do] DOI:10.1186/s13104-017-2447-6


  10 / 3913 MEDLINE  
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[PMID]:28209381
[Au] Autor:Suiwal S; Kiefer G; Schmitz F; Schwarz K
[Ad] Endereço:Saarland University, Department of Neuroanatomy, Institute of Anatomy and Cell Biology, Kirrbergerstrasse, 66421 Homburg/Saar, Germany.
[Ti] Título:An easy, fast and "low-tech"-equipment-requiring alternative method to optimize immunolabelling conditions for pre-embedding immunogold electron microscopy and to correlate light and electron microscopical immunogold labelling results.
[So] Source:J Immunol Methods;444:7-16, 2017 May.
[Is] ISSN:1872-7905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Correlating light microscopic immunolabelling results with electron microscopic data is of great interest in many fields of biomedical research but typically requires very specialized, expensive equipment and complex procedures which are not available in most labs. In this technical study, we describe an easy and "low-tech"-equipment-requiring pre-embedding immunolabelling approach that allows correlation of light microscopical immunolabelling results with electron microscopic (EM) data as demonstrated by the example of immunolabelled synaptic ribbons from retinal rod photoreceptor synapses. This pre-embedding approach does not require specialized embedding devices but only commonly available equipment. The cryostat section-based procedure allows optimization of the pre-embedding immunolabelling conditions at the less laborious and time-consuming light microscopic (LM) level before the ultrastructural analyses of the immunolabelled structures can be performed on the same sample after ultrathin sectioning without further modification. The same photoreceptor synapse that has been first studied at the light microscopic level can be subsequently analyzed with this approach at the electron microscopic level at individual ultrathin sections or serial ultrathin sections from individual, identical synapses. Higher resolution EM analyses of the immunolabelled synapses can be performed with only minor modifications of the combined LM/EM procedure. The detergent-free procedure is applicable even for weakly fixed cryostat sections which is a relevant aspect for many antibodies that do not work with more strongly fixed biological samples.
[Mh] Termos MeSH primário: Sinapses Elétricas/imunologia
Sinapses Elétricas/ultraestrutura
Interpretação de Imagem Assistida por Computador/métodos
Imuno-Histoquímica/métodos
Microscopia Eletrônica de Transmissão/métodos
Células Fotorreceptoras Retinianas Bastonetes/imunologia
Células Fotorreceptoras Retinianas Bastonetes/ultraestrutura
Inclusão do Tecido/métodos
[Mh] Termos MeSH secundário: Animais
Bovinos
Proteínas do Olho/imunologia
Interpretação de Imagem Assistida por Computador/instrumentação
Imuno-Histoquímica/instrumentação
Microscopia Eletrônica de Transmissão/instrumentação
Microtomia
Fixação de Tecidos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Eye Proteins)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170713
[Lr] Data última revisão:
170713
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170218
[St] Status:MEDLINE



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