Agradecemos Sonja Dufentester e Elke Weber para assistência técnica especializada. Agradecemos Faekah Gohar pela revisão do manuscrito e Stefan Brückner para suporte informática médica. Este trabalho foi apoiado por uma bolsa interdisciplinar do Else-Kröner-Fresenius-Stiftung (2012_A94). D. Bettenworth foi apoiado por uma bolsa de pesquisa da Faculdade de Medicina da Westfälische Wilhelms-Universität Münster. M. Brückner foi apoiado por uma &quot;Gerok&quot; posição de rotação da Deutsche Forschungsgemeinschaft (DFG SFB1009B8). Agradecemos Heike Blum para ilustração do desenho animado mouse.

Todos os experimentos com animais foram aprovados pelo Landesamt für Natur, Umwelt und Verbraucherschutz (LANUV) de acordo com a Lei de Proteção de Animais alemão. 1. Materiais e montagem de experimento <ol><li> Cuidados com os animais <ol><li> Use feminino ou camundongos machos de qualquer tensão pesando 20 a 25 g e abrigá-los de acordo com a legislação cuidado animal local. </li><li> Alimentar ratos com comida para roedores e especial aplicar ração isenta de alfafa pelo menos três dias antes da fluorescência exames para minimizar endoluminal auto-fluorescência. </li><li> Fornecer autoclavado beber água ad libitum. </li></ol></li><li> A indução da colite induzida por DSS aguda <ol><li> Prepara-se uma 3% (w / v) de sulfato de sódio de dextrano (DSS, peso molecular: 36,000-50,000 Da) solução por dissolução de 3 g de DSS em 100 ml de água autoclavada. Oferecer esta solução como a água potável exclusiva para ad libitum camundongos e calcular 5 ml de solução por DSS-mouse / dia. Alimentação contcamundongos rol com água autoclavada sem DSS ad libitum 9. </li></ol></li><li> A indução de câncer colorretal <ol><li> Dissolver azoximetano mutagénicas (OMA) (CUIDADO Pode causar cancro e defeitos genéticos!) Em solução salina isotónica estéril para se obter uma concentração final de 1 mg / ml. Aplicar uma dose única de 10 mg por kg de peso corporal OMA intra-peritoneal com uma seringa de 1 ml (30 g) 10. </li><li> Camundongos desafio (excluindo os ratos de controle) com ciclos repetitivos de 3% (w / v) DSS do dia 0-7, dia 14-21, dia 28-35 e dia 42 a 49 de induzir inflamatória impulsionado cancerogenesis colorretal. Alimentar ratos com água esterilizada apenas entre esses desafios (ver Figura 4A por um cronograma detalhado). Alimentar ratos de controle em água esterilizada durante todo o experimento. </li></ol></li><li> Preparação de fluorescência endoscopia (FE) <ol><li> Use Fluorescein-Isothiocyanat (ITCF) - dextrano (peso molecular de 70.000 Da; FITC: glicose = 1: 250) para detecção de adenoma do cólon por realce visual de displasia associada padrão vascular. </li><li> Administrar 60 mg de dextrano conjugado com FITC diluído em 100 ul de PBS por via intravenosa 5 minutos antes da análise de fluorescência endoscópica. </li></ol></li><li> Anestesia <ol><li> Fornecer um suprimento contínuo para anestesia de isoflurano (1,5 LO 2 / min; 1,5-2% em volume de isoflurano [2-cloro-2-(difluorometoxi) -1,1,1-trifluoro-etano]). Use equipamentos especiais anestesia veterinária, com uma máscara para controlar firmemente a anestesia. </li></ol></li><li> Preparação de enema <ol><li> Instilar 2 ml de fluido de enema (conteúdo: hidrogenofosfato dissódico 1,5% (w / v) e di-hidrogenofosfato de sódio a 11% (w / v)) para o cólon, se se suspeita de significativa fecal que podem obscurecer a visão. </li></ol></li></ol> 2 Equipamento técnico <ol><li> Use uma estação de trabalho endoscópica veterinária que é desenvolvida e aprovada para o uso de animais pequenos endoscopia. Conecte o workstation para uma unidade de câmera, uma fonte de luz xenon, uma bomba de ar e um monitor convencional PC para luz branca endoscopia. Em seguida, conecte a câmera eo telescópio rígido em miniatura (diâmetro externo 1,9 milímetros, 10 cm de comprimento; Figura 5). </li><li> Use bainha endoscópio com canal de trabalho (Figura 5D) para a aplicação de pinça de biópsia ou tubo de injecção. Use a bainha sem canal de trabalho para colonoscopia diagnóstica. </li><li> Defina as configurações da fonte de luz para fluorescência endoscopia para excitar traçadores utilizado (por exemplo, 490 nm para dextrano FITC-conjugado). Além disso, a integração de um filtro de passagem de banda adequada entre o telescópio e a câmara (por exemplo, 525 nm para o dextrano conjugado com FITC). </li><li> Insira uma pinça de biópsia flexível (3, 28. Charr cm) através do canal de trabalho do endoscópio para obter biópsias. </li><li> Introduzir tubo de injecção flexível (0,96 mm), através do canal de trabalho para tópica, intramucosal ou Endoluminal umdministração de agentes de diagnóstico ou terapêuticos. </li><li> Use uma mesa de exame aquecida com uma temperatura de 42 ° C. Isso evita que os ratos se tornando hipotermia durante o exame. </li></ol> 3 Anestesia de Animais <ol><li> Coloque rato numa caixa à prova de fugas, mas pequena e administrar isoflurano (100% (v / v), 5% em volume, de 3 L / min). Espere até que o mouse perde a consciência. </li><li> Transferir o mouse sobre a mesa de exame de endoscopia. Continue inalação isoflurano via máscara facial com uma dose de 100% v / v, 1,5 vol%, 1,5 L / min. Sempre aplique pomada para evitar o ressecamento dos olhos e sob anestesia. </li><li> Avaliar a eficácia da anestesia, verificando os reflexos. Verifique a &#39;volta em torno do reflexo &quot;: se suficientemente anestesiado, um rato que coloca em suas costas não deve virar. Confira os &#39;pés reflex &quot;: quando a anestesia é adequado, macio apertando entre os dedos do animal não deve conduzir à retirada da perna (fase detolerância cirúrgico). </li></ol> 4. Colonoscopia <ol><li> Coloque rato anestesiado propenso / de costas na mesa de exame. </li><li> Administrar 2 ml de enema através de uma cânula abotoado no cólon se houver suspeita de acúmulo fecal significativo que pode obscurecer a visão. Espere por ratos para defecar após a administração do enema. Insira o endoscópio com muito cuidado para evitar a perfuração. </li><li> Abrir ambas as válvulas da bainha com um deles estar ligado à bomba de ar. Selar a outra válvula com o dedo indicador para dispensar o ar. Inflar cólon com ar, lentamente e com cuidado, em especial no caso de biópsia ou de injecção. </li><li> Avance endoscópio apenas até a articulação do cólon direito de evitar a perfuração (4-5 cm de ânus). </li><li> Colonoscopia diagnóstica <ol><li> Examine a mucosa por alterações inflamatórias ou malignas, enquanto puxa de volta o endoscópio. Puxar lentamente para avaliar toda a circunferência do intestino. Avaliar pa intraluminalthologies utilizando sistemas de pontuação estabelecidos apropriados necessários. </li><li> Para justificar a posição endoscópica idêntico para a aquisição de imagens durante as visualizações repetitivas das áreas das feridas, observe a distância entre o ânus murino ea lesão da mucosa. Além disso, utilizar a ponta da pinça de biópsia como um espaçador, para atingir a distância idêntica entre o endoscópio e a área da ferida durante a aquisição de imagem. O tamanho da ferida está relacionada com o tamanho da bainha de endoscópio, que é composta por 3 mm. NOTA: Coloque endoscópio em posição idêntica em comparação óptica com documentação fotográfica dos exames anteriores. Lesões de medidas no mesmo ângulo e distância em cada seguimento exame endoscópico. </li></ol></li><li> Procedimento de biópsia <ol><li> Tome biópsias com a ajuda de dois investigadores. Introduzir uma pinça de biópsia com cuidado através do canal de trabalho até que a ponta da pinça é visível no monitor para o segundo investigador. Fórceps Abra e feche com cuidado para aperfuração vazio. </li><li> Mover uma pinça para o local da patologia. </li></ol></li><li> Procedimento de injeção <ol><li> Executar um procedimento de injecção, com a ajuda de dois investigadores. Pré-enchimento do tubo flexível de injecção (0,96 mm) completamente com o agente a ser administrado. Empurre tubo através do canal de trabalho até que a cânula (30 G) é visível no monitor para o segundo investigador. Preparar seringa fino e suavemente administrar a quantidade de agente de diagnóstico ou terapêutico requerido. Os volumes de injecção deve ser de 50 máximo l. </li><li> Insira a agulha no submocosa em um ângulo de 15-30 graus. Encare o chanfro na direção da mucosa. A mucosa mostra um sinal de elevação característica após a injeção de sucesso. </li></ol></li><li> Fluorescência endoscopia (FE) <ol><li> Administrar 60 mg de dextrano conjugado com FITC diluído em 100 ul de PBS por via intravenosa antes fluorescência exame endoscópico. </li><li> Verifique o ponto de tempo ideal entre a injeção deo traçador fluorescente marcado e o procedimento de imagem o qual é dependente do traçador farmacologia. Configurar as definições de sistema de filtro de banda de acordo com o comprimento de onda de excitação e emissão do traçador utilizado. Realizar endoscopia para marcadores de fluorescência de volume de sangue não-específicos (por exemplo, FITC), imediatamente após a injecção intravenosa do corante fluorescente para avaliar o padrão vascular da superfície da mucosa. </li><li> Considere imaginando várias horas após a aplicação do traçador no caso dos marcadores direcionados ou &quot;sondas inteligentes&quot; para proporcionar uma melhor alvo para fundo de razão. </li><li> Realizar foto e vídeo-documentação dos resultados. </li></ol></li></ol> 5 Pós-colonoscopia <ol><li> Separe o rato em uma gaiola vazia e coloque-a sobre uma toalha de papel para proteger o mouse de aspiração de areia. Aqueça o mouse com uma lâmpada de luz vermelha para evitar a hipotermia. Observe o mouse e não deixe sem vigilância até que recuperou suficienteconsciência para manter decúbito esternal. Uma vez completamente consciente, coloque o mouse de volta para sua respectiva gaiola. </li><li> No final da experiência, o lugar do rato numa caixa à prova de fugas, mas pequena e administrar CO 2 (100% (v / v), 100% em volume, de 3 L / min). Espere até que o mouse perde a consciência completa e pára de respirar. Despacho do mouse por fratura do colo. Realizar laparotomia eo explantar cólon. Abra o cólon longitudinalmente e lavá-lo para posterior avaliação histológica ou molecular. </li></ol>

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Os autores não têm nada a revelar.

Cicatrização do epitélio é um processo contínuo. Esfoliação fisiológica contínua de células superficiais da mucosa gastrointestinal ocorre exigindo a regeneração frequente de células epiteliais 16. Consequentemente, a cicatrização de feridas prejudicada tem um imenso impacto sobre várias doenças, incluindo úlceras gastrointestinais e vazamento da anastomose 17 18. Avaliação da base molecular, assim como potenciais candidatos a medicamentos para estimular a cicatrização, só po...

Modelos animais têm muito enriquecido nossa compreensão de inúmeras patologias intestinais. O rato de laboratório (Mus musculus) surgiu como um modelo animal primordial na investigação biomédica devido à sua informação genética e genômica abundante e está prontamente disponível em linhagens transgênicas e mata-mata. Além de reforçar a compreensão patogênese da doença, modelos animais também são usados ​​principalmente para testar drogas candidatos, bem como intervenções diagnósticas ou terapêuticas pré-clínicos. No entanto, apesar da variedade de modelos de ratos, simulando a doença humana, muitas opções de diagnóstico e de intervenção que são rotineiramente usados ​​no tratamento do paciente não estão disponíveis para os ratos. Deste modo, as estratégias de vigilância para controlar o curso da doença de murino ou o efeito de intervenções terapêuticas são geralmente limitados para observações indirectas ou post mortem análises. Embora existam procedimentos não-invasivos para os ratos de monitoramento vitalidade como índices de atividade da doença, quantification de perda de peso ou ganho, sangue, urina e fezes análises, estas são apenas indicadores indiretos e são influenciados pela variabilidade inter-individual. Além disso, análises post mortem evitar observações longitudinais em momentos repetitivos. Técnicas de imagem sofisticados para monitorar a atividade da doença em camundongos só recentemente foram introduzidos 1,2. Embora essas técnicas de imagem permitem análises repetitivas, eles apenas fornecem uma visão descritiva e muitas vezes imprecisas sobre o intestino, não permitir a visualização da mucosa direta ou permitir intervenções diagnósticas ou terapêuticas, como a aquisição de biópsia ou aplicação tópica e intramucosal de candidatos a fármacos. Recentemente, os sistemas de endoscopia de alta resolução para utilização em ratos vivos foram desenvolvidos 3,4. Pela primeira vez, essas técnicas endoscópicas permitir a visualização direta de patologias de doenças do cólon endoluminal como cicatrização de feridas ou inflamação intestinal proporcionando objective estado, em tempo real, permitindo estudos longitudinais no mesmo animal em intervalos de tempo repetitivos. Além de permitir que as biópsias repetidas em um ratinho individual, sistemas de endoscopia também pode ser utilizado para influenciar terapeuticamente um tumor distinta ou inflamação localizada, permitindo a aplicação directa de uma substância para a área de interesse. Além disso, como substâncias terapêuticas e de controlo podem ser entregues directamente para a área de interesse, esta pode ser realizada no mesmo ratinho, não incluindo a variabilidade inter-individual. Estes sistemas de terem sido utilizadas para a avaliação da inflamação do cólon, cicatrização de feridas, biópsias hepáticas laparoscópicas e indução ortotópico de tumores hepáticos 8 e desenvolvimento do tumor através de vários sistemas de pontuação, tais como o índice endoscópica murino de gravidade da colite (MEICS) 5-7. MEICS consiste de cinco parâmetros para avaliar a inflamação: espessamento da parede do cólon, alterações do padrão vascular, presença de fibrina, a granularidade dos Mucossuperfície al, e fezes de consistência. Neste protocolo, descrevem o uso de endoscopia rígida em modelos murinos de cicatrização intestinal, inflamação e câncer de cólon. Em primeiro lugar, demonstrar a avaliação endoscópica da cicatrização e inflamação do cólon, bem como a avaliação longitudinal da atividade colite eo estudo da cancerogenesis no cólon murino. Além do uso descritivo da endoscopia murino, nós fornecemos instruções detalhadas sobre o uso de instrumentos endoscópicos para obter biópsias, bem como a aplicação tópica e intramucosal de diferentes componentes de interesse (por exemplo, candidatos a fármacos ou células tumorais). Finalmente, demonstramos a utilização de endoscopia fluorescência murino, que emprega técnicas sofisticadas de imagens moleculares, na definição de tumores colo-rectais.

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		<tr height="20">
			<td height="20" style="height:20px;width:469px;"></td>
			<td style="width:423px;"></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Reagents</td>
			<td style="width:423px;"></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Alfalfa-free diet</td>
			<td>Harlan Laboritories, Madison, USA</td>
			<td style="width:320px;">2014</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Azoxymethane (AOM)</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">A5486</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bepanthen eye ointment</td>
			<td style="width:423px;">Bayer, Leverkusen, Germany</td>
			<td style="width:320px;">80469764</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dextran sulphate sodium (DSS)</td>
			<td>TdB Consulatancy, Uppsala, Sweden</td>
			<td>DB001</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Eosin</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 4382</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Ethylenediaminetetraacetic acid (EDTA)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 9884</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Falcon Tube 50ml</td>
			<td>BD Biosciences, Erembodegem, Belgium</td>
			<td style="width:320px;">352070</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Florene 100V/V</td>
			<td>Abbott, Wiesbaden, Germany</td>
			<td style="width:320px;">B506</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fluorescein-Isothiocyanat (FITC)&#8211;dextrane&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">FD4-250MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Haematoxylin&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">HHS32-1L</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Isopentane (2- Methylbutane)</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">M32631-1L</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Methylene blue</td>
			<td style="width:423px;">Merck, Darmstadt, Germany</td>
			<td style="width:320px;">1159430025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">O.C.T. Tissue Tek compound&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sakura, Zoeterwonde, Netherlands</td>
			<td style="width:320px;">4583</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Omnican F - canula</td>
			<td style="width:423px;">Braun, Melsungen, Germany</td>
			<td style="width:320px;">9161502</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Phosphate buffered saline, PBS</td>
			<td style="width:423px;">Lonza, Verviers, Belgium</td>
			<td style="width:320px;">4629</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Sodium Chloride 0,9%</td>
			<td>Braun, Melsungen, Germany</td>
			<td style="width:320px;">5/12211095/0411</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Standard diet</td>
			<td>Altromin, Lage, Germany</td>
			<td style="width:320px;">1320</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tissue-Tek Cryomold</td>
			<td>Sakura, Leiden, Netherlands</td>
			<td style="width:320px;">4566</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Vitro &#8211; Clud&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">R. Langenbrinck, Teningen, Germany</td>
			<td style="width:320px;">04-0002&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Equipment</td>
			<td></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">AIDA Control</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 096020</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bandpass filter</td>
			<td>Semrock, Rochester, USA</td>
			<td style="width:320px;">HC 716/40</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bandpass filter</td>
			<td>Semrock, Rochester, USA</td>
			<td style="width:320px;">HC 809/81</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Biopsy Forceps, 3 Fr., 28cm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61071ZJ</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dell Monitor</td>
			<td>Dell, Frankfurt am Main, Germany</td>
			<td style="width:320px;">U2412Mb</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Examination Sheath, 9 Fr.</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61029D</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Examination Sheath, 9 Fr.</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61029C</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fiber Optic Light Cable, 3.5mm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69495NL</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fluorescein Blue Filter System</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20100032</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fluorescein Barrier Filter</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20100033</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Foot switch</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20010430</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">HOPKINS Telescope, 1.9mm, Length 10cm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">1830231</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SCB D-light P&#160;</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 133720</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SCB tricam SL II</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 2230 20</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tubing set instruments VETPUMP II</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69811</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tricam PDD PAL</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20221037</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">UniVet Porta</td>
			<td>Groppler Medizintechnik, Deggendorf, Germany</td>
			<td style="width:320px;">BKGM 0451</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Vetpump 2</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69321620</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine endoscopy for in vivo multimodal imaging of carcinogenesis and assessment of intestinal wound healing and inflammation

Modelos de mouse são amplamente utilizados para estudar patogênese de doenças humanas e para avaliar os procedimentos de diagnóstico, bem como intervenções terapêuticas pré-clínicos. No entanto, a avaliação válida de alterações patológicas, muitas vezes requer análise histológica, e quando realizado ex vivo, necessita de morte do animal. Portanto, em situações experimentais convencionais, exames de acompanhamento intra-individuais são raramente possível. Assim, o desenvolvimento de endoscopia de murino em ratinhos vivos investigadores permite pela primeira vez a ambos visualizar diretamente a mucosa gastrointestinal e também repetir o processo para a monitorização de alterações. Existem inúmeras aplicações para in vivo murino endoscopia, incluindo estudar inflamação intestinal ou cicatrização de feridas, a obtenção de biópsias da mucosa repetidamente, e localmente administrar agentes de diagnóstico ou terapêuticos em uso de cateter de injeção em miniatura. Mo de diagnóstico por imagem, mais recentemente, a imagem molecular tem estendidodalidades permitindo a detecção específica de moléculas alvo específicas distintas, usando as fotossondas. Em conclusão, a endoscopia murino surgiu como uma nova tecnologia de ponta para o diagnóstico experimental in vivo de imagens e pode ter um impacto significativo em pesquisas pré-clínicas em vários campos.

Na monitorização in vivo da cicatrização do intestino Durante a endoscopia de rotina, feridas na mucosa foram induzidas mecanicamente por uma pinça de biópsia em miniatura com um diâmetro de 3 Francês (igual a 1 mm; Figura 1A). Subsequentemente, a cicatrização de feridas foi monitorada por exames endoscópicos diários e quantificada por medição da área de ferida residual utilizando o software de edição, por exemplo, o ImageJ (Fig...

Watch this Scientific Journal Video about Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation at JoVE.com

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

Murino Endoscopia para<em&gt; In Vivo</em&gt; Cura Multimodal imagem de Carcinogênese e Avaliação de Intestinal ferida e inflamação

Pequenas técnicas de imagem animais permitem exames de diagnóstico de série e intervenções terapêuticas in vivo. Recentemente, o leque de aplicações tem aumentado significativamente e atualmente inclui avaliação do desenvolvimento de tumor do cólon, cicatrização de feridas e monitoramento da inflamação. Este protocolo ilustra essas aplicações potenciais diferentes de endoscopia murino.

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions ...

murine-endoscopy-for-vivo-multimodal-imaging-carcinogenesis

Research

JoVE Journal

Medicine

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

18.5K visualizações.  University Hospital Münster.  Pequenas técnicas de imagem animais permitem exames de diagnóstico de série e intervenções terapêuticas in vivo. Recentemente, o leque de aplicações tem aumentado significativamente e atualmente inclui avaliação do desenvolvimento de tumor do cólon, cicatrização de feridas e monitoramento da inflamação. Este protocolo ilustra essas aplicações potenciais diferentes de endoscopia murino. 

Vídeo: Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading to an accumulation of lipid-laden intra-luminal plaque, smooth muscle cell proliferation and progressive narrowing of the vessel lumen. The formation of such vulnerable plaques prone to rupture underlies the majority of cases of acute myocardial infarction. The complex molecular and cellular inflammatory cascade is orchestrated by the recruitment of T lymphocytes and macrophages and their paracrine effects on endothelial and smooth muscle cells.1
Molecular imaging in atherosclerosis has evolved into an important clinical and research tool that allows in vivo visualization of inflammation and other biological processes. Several recent examples demonstrate the ability to detect high-risk plaques in patients, and assess the effects of pharmacotherapeutics in atherosclerosis.4 While a number of molecular imaging approaches (in particular MRI and PET) can image biological aspects of large vessels such as the carotid arteries, scant options exist for imaging of coronary arteries.2 The advent of high-resolution optical imaging strategies, in particular near-infrared fluorescence (NIRF), coupled with activatable fluorescent probes, have enhanced sensitivity and led to the development of new intravascular strategies to improve biological imaging of human coronary atherosclerosis.
Near infrared fluorescence (NIRF) molecular imaging utilizes excitation light with a defined band width (650-900 nm) as a source of photons that, when delivered to an optical contrast agent or fluorescent probe, emits fluorescence in the NIR window that can be detected using an appropriate emission filter and a high sensitivity charge-coupled camera. As opposed to visible light, NIR light penetrates deeply into tissue, is markedly less attenuated by endogenous photon absorbers such as hemoglobin, lipid and water, and enables high target-to-background ratios due to reduced autofluorescence in the NIR window. Imaging within the NIR 'window' can substantially improve the potential for in vivo imaging.2,5
Inflammatory cysteine proteases have been well studied using activatable NIRF probes10, and play important roles in atherogenesis. Via degradation of the extracellular matrix, cysteine proteases contribute importantly to the progression and complications of atherosclerosis8. In particular, the cysteine protease, cathepsin B, is highly expressed and colocalizes with macrophages in experimental murine, rabbit, and human atheromata.3,6,7 In addition, cathepsin B activity in plaques can be sensed in vivo utilizing a previously described 1-D intravascular near-infrared fluorescence technology6, in conjunction with an injectable nanosensor agent that consists of a poly-lysine polymer backbone derivatized with multiple NIR fluorochromes (VM110/Prosense750, ex/em 750/780nm, VisEn Medical, Woburn, MA) that results in strong intramolecular quenching at baseline.10 Following targeted enzymatic cleavage by cysteine proteases such as cathepsin B (known to colocalize with plaque macrophages), the fluorochromes separate, resulting in substantial amplification of the NIRF signal. Intravascular detection of NIR fluorescence signal by the utilized novel 2D intravascular NIRF catheter now enables high-resolution, geometrically accurate in vivo detection of cathepsin B activity in inflamed plaque.
In vivo molecular imaging of atherosclerosis using catheter-based 2D NIRF imaging, as opposed to a prior 1-D spectroscopic approach,6 is a novel and promising tool that utilizes augmented protease activity in macrophage-rich plaque to detect vascular inflammation.11,12 The following research protocol describes the use of an intravascular 2-dimensional NIRF catheter to image and characterize plaque structure utilizing key aspects of plaque biology. It is a translatable platform that when integrated with existing clinical imaging technologies including angiography and intravascular ultrasound (IVUS), offers a unique and novel integrated multimodal molecular imaging technique that distinguishes inflammatory atheromata, and allows detection of intravascular NIRF signals in human-sized coronary arteries.

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

In vivo Near Infrared fluorescência de imagem intravascular (NIRF) Molecular da placa inflamatória, uma Abordagem Multimodal para imagem da Aterosclerose

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

Medicina

Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the complex cellular and molecular of intestinal pathology. Although the tunica mucosa is often the organ of interest in many inflammatory diseases, recent works demonstrated that the muscularis externa (ME) is also a highly immunocompetent organ that harbours a dense network of resident immunocytes.1,2 These works were performed within the standardized model of intestinal manipulation (IM) that leads to inflammation of the bowel wall, mainly limited to the ME. Clinically this inflammation leads to prolonged intestinal dysmotility, known as postoperative ileus (POI) which is a frequent and unavoidable complication after abdominal surgery.3 The inflammation is characterized by liberation of proinflammatory mediators such as IL-64 or IL-1&beta; or inhibitory neurotransmitters like nitric oxide (NO).5 Subsequently, tremendous numbers of immunocytes extravasate into the ME, dominated by polymorphonuclear neutrophils (PMN) and monocytes and finally maintain POI.2 Lasting for days, this intestinal paralysis leads to an increased risk of aspiration, bacterial translocation and infectious complications up to sepsis and multi organ failure and causes a high economic burden.6
In this manuscript we demonstrate the standardized model of IM and in vivo assessment of gastrointestinal transit (GIT) and colonic transit. Furthermore we demonstrate a method for separation of the ME from the tunica mucosa followed by immunological analysis, which is crucial to distinguish between the inflammatory responses in these both highly immunoactive bowel wall compartments. All analyses are easily transferable to any other research models, affecting gastrointestinal function.

Postoperative ileus (POI) is a complication of abdominal surgery leading to increased morbidity and a prolonged hospital stay. Because prophylactic or therapeutic strategies are lacking intensified research is necessary. Therefore we established a standardized and feasible mouse model to investigate the pathophysiology of POI and to study potential therapeutic options.

Avaliação Funcional da motilidade intestinal e inflamação da parede do intestino em Roedores: Análises em um modelo padronizado de Manipulação Intestinal

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Modelo Murino de Cicatrização

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

Um modelo murino de irreversível e reversível obstrução ureteral unilateral

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Multimodal Quantitative Phase Imaging with Digital Holographic Microscopy Accurately Assesses Intestinal Inflammation and Epithelial Wound Healing

The incidence of inflammatory bowel disease, i.e., Crohn&#39;s disease and Ulcerative colitis, has significantly increased over the last decade. The etiology of IBD remains unknown and current therapeutic strategies are based on the unspecific suppression of the immune system. The development of treatments that specifically target intestinal inflammation and epithelial wound healing could significantly improve management of IBD, however this requires accurate detection of inflammatory changes. Currently, potential drug candidates are usually evaluated using animal models in vivo or with cell culture based techniques in vitro. Histological examination usually requires the cells or tissues of interest to be stained, which may alter the sample characteristics and furthermore, the interpretation of findings can vary by investigator expertise. Digital holographic microscopy (DHM), based on the detection of optical path length delay, allows stain-free quantitative phase contrast imaging. This allows the results to be directly correlated with absolute biophysical parameters. We demonstrate how measurement of changes in tissue density with DHM, based on refractive index measurement, can quantify inflammatory alterations, without staining, in different layers of colonic tissue specimens from mice and humans with colitis. Additionally, we demonstrate continuous multimodal label-free monitoring of epithelial wound healing in vitro, possible using DHM through the simple automated determination of the wounded area and simultaneous determination of morphological parameters such as dry mass and layer thickness of migrating cells. In conclusion, DHM represents a valuable, novel and quantitative tool for the assessment of intestinal inflammation with absolute values for parameters possible, simplified quantification of epithelial wound healing in vitro and therefore has high potential for translational diagnostic use.

Accurate assessment of anti-inflammatory effects is of utmost importance for the evaluation of potential new drugs for the treatment of inflammatory bowel disease. Digital holographic microscopy provides assessment of inflammation in murine and human colonic tissue samples as well as automated multimodal evaluation of epithelial wound healing in vitro.

Multimodal Quantitative Fase de imagem com o Digital Holographic Microscopia com precisão Avalia Cura inflamação intestinal e epitelial de Feridas

The incidence of inflammatory bowel disease, i.e., Crohn&#39;s disease and Ulcerative colitis, has significantly increased over the last decade. The ...

Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not routinely employed in animal models. Conventional experimental readouts often rely on post mortem and ex vivo analyses, which prevent intra-individual follow-up examinations and increase the number of study animals needed. Fluorescence-mediated tomography enables the non-invasive, repetitive, quantitative, three-dimensional assessment of fluorescent probes. It is highly sensitive and permits the use of molecular makers, which allows for the specific detection and characterization of distinct molecular targets. In particular, targeted probes represent an innovative tool for analyzing gene activation and protein expression in inflammation, autoimmune disease, infection, vascular disease, cell migration, tumorigenesis, etc. In this article, we provide step-by-step instructions on this sophisticated imaging technology for the in vivo detection and characterization of inflammation (i.e., F4/80-positive macrophage infiltration) in a widely used murine model of intestinal inflammation. This technique might also be used in other research areas, such as immune cell or stem cell tracking.

Target-specific probes represent an innovative tool for analyzing molecular mechanisms, such as protein expression in various types of disease (e.g., inflammation, infection, and tumorigenesis). In this study, we describe a quantitative three-dimensional tomographic assessment of intestinal macrophage infiltration in a murine model of colitis using F4/80-specific fluorescence-mediated tomography.

Tomografia computadorizada mediada por fluorescência para a detecção e quantificação de inflamação Intestinal murino macrófago-relacionados

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not ...

Microscopy Based Methods for the Assessment of Epithelial Cell Migration During In Vitro Wound Healing

Cell migration is a mandatory aspect for wound healing. Creating artificial wounds on research animal models often results in costly and complicated experimental procedures, while potentially lacking in precision. In vitro culture of epithelial cell lines provides a suitable platform for researching the cell migratory behavior in wound healing and the impact of treatments on these cells. The physiology of epithelial cells is often studied in non-confluent conditions; however, this approach may not resemble natural wound healing conditions. Disrupting the epithelium integrity by mechanical means generates a realistic model, but may impede the application of molecular techniques. Consequently, microscopy based techniques are optimal for studying epithelial cell migration in vitro. Here we detail two specific methods, the artificial wound scratch assay and the artificial migration front assay, that can obtain quantitative and qualitative data, respectively, on the migratory performance of epithelial cells.

Microscopy Based Methods for the Assessment of Epithelial Cell Migration During  In Vitro Wound Healing

This manuscript describes how incision-like lesions made on cultured epithelial cell monolayers conveniently model wound healing in vitro, allowing for imaging by confocal or laser scanning microscopy, and which can provide high-quality quantitative and qualitative data for studying both cell behavior and the mechanisms involved in migration.

Microscopia com base em métodos de avaliação da migração de células epiteliais durante a cicatrização em Vitro

Cell migration is a mandatory aspect for wound healing. Creating artificial wounds on research animal models often results in costly and complicated ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

Cicatrização de feridas abrasão epitelial da córnea com Burr Ocular como um modelo para córnea

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two dimensional (2D) and three dimensional (3D) culture has generated a wealth of information not only about corneal biology but also about wound healing, myofibroblast biology, and scarring in general. The goal of the protocol is an assay system for quantifying myofibroblast development, which characterizes scarring. We demonstrate a corneal organ culture ex vivo model using pig eyes. In this anterior keratectomy wound, corneas still in the globe are wounded with a circular blade called a trephine. A plug of approximately 1/3 of the anterior cornea is removed including the epithelium, the basement membrane, and the anterior part of the stroma. After wounding, corneas are cut from the globe, mounted on a collagen/agar base, and cultured for two weeks in supplemented-serum free medium with stabilized vitamin C to augment cell proliferation and extracellular matrix secretion by resident fibroblasts. Activation of myofibroblasts in the anterior stroma is evident in the healed cornea. This model can be used to assay wound closure, the development of myofibroblasts and fibrotic markers, and for toxicology studies. In addition, the effects of small molecule inhibitors as well as lipid-mediated siRNA transfection for gene knockdown can be tested in this system.

A protocol for an ex vivo corneal organ culture model useful for wound healing studies is described. This model system can be used to assess the effects of agents to promote regenerative healing or drug toxicity in an organized 3D multicellular environment.

Ex Vivo modelo de cultura órgão da córnea para cicatrização de feridas estudos

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two ...

Targeting Gray Rami Communicantes in Selective Chemical Lumbar Sympathectomy

Chemical lumbar sympathectomy (CLS) is a commonly used, minimally invasive procedure for the treatment of conditions including ischemic diseases of the lower extremities, hyperhidrosis, etc. It is commonly practiced to position the puncture needle tip in front of the anterior fascia of the psoas major muscle and inject the inactivating agent around the sympathetic trunk, which is defined as conventional CLS. Although relatively rare, ureteropelvic damage is the most frequently reported complication of conventional CLS and can cause serious harm to patients. We found that injecting the inactivating agent behind the anterior fascia, which only targets gray rami communicantes, helped achieve therapeutic efficacy in&#160;vasodilation, sweat reduction, and pain relief comparable to conventional CLS, and serious complications were largely reduced. We define this procedure as selective CLS. Here, we present a protocol of selective CLS. The precise needle tract and accurate evaluation of the spreading of the contrast agent are critical to ensure that the drug is injected behind the anterior fascia of the psoas major muscle. The needle tip is at approximately one-third the dividing line of the vertebral body in the lateral view of a lumbar X-ray. The contrast is mainly confined around the needle tip and spreads outward and downward along the psoas muscle fibers. In this way, the anterior fascia provides a natural barrier for the ureteropelvic area, and the psoas major muscle provides a natural barrier for the lumbar nerve root. There are several highlights of this article, including 1) a detailed description of the selective CLS procedures, 2) an explanation of the anatomical basis for the implementation of selective CLS, and 3) an explanation of the differences between selective and conventional CLS.

We present a protocol for selective chemical lumbar sympathectomy (CLS), which inactivates only gray rami communicantes and not the sympathetic trunk. Selective CLS can help achieve therapeutic efficacy in vasodilation, sweat reduction, and pain relief that are comparable to conventional CLS, and serious complications, particularly ureteropelvic damages, can be reduced.

Direcionamento de Rami cinza Communicantes em simpatectomia lombar química seletiva

Chemical lumbar sympathectomy (CLS) is a commonly used, minimally invasive procedure for the treatment of conditions including ischemic diseases of the ...