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<table><tbody><tr><td>10% Tween-20</td><td>Bio-Rad Laboratories</td><td>1662404</td><td>NA</td></tr><tr><td>20x PBS, pH7.4</td><td>Growcells.com</td><td>MRGF-6396</td><td>2,74M NaCl, 53,7 mM KCl, 20,3 mM Na2HPO4, 29,4 mM KH2PO4&nbsp;</td></tr><tr><td>20x SSC</td><td>Quality Biological</td><td>351-003-131</td><td>3M NaCl, 0,3M Sodium Citrate</td></tr><tr><td>37 &deg;C incubator</td><td>Fisher Scientific (Isotemp)</td><td>15-015-2633</td><td>NA</td></tr><tr><td>37 &deg;C water bath</td><td>Fisher Scientific</td><td>FSGPD15D</td><td>NA</td></tr><tr><td>Aluminum foil</td><td>Reynolds Wrap/Fisher Brand</td><td>15-078-291</td><td>NA</td></tr><tr><td>Antibody: Mouse-anti-Abrupt</td><td>DSHB</td><td>Abrupt; RRID: AB_528344&nbsp;</td><td>Dilution: 1:20</td></tr><tr><td>Antibody: Chicken anti-GFP</td><td>Millipore Sigma</td><td>Catalog# 06-896; RRID: AB-619712</td><td>Dilution: 1:500</td></tr><tr><td>Antibody: Rabbit anti-GFP</td><td>Life Technologies</td><td>#11122; RRID: AB_221569</td><td>Dilution: 1:500</td></tr><tr><td>Antibody: Rabbit anti-Dll</td><td>Genscript/Desplan lab</td><td>-</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Rat anti-NCadherin</td><td>DSHB</td><td>DN-Ex #8, RRID: AB_528121</td><td>Dilution: 1:50</td></tr><tr><td>Antibody: Rabbit anti-SoxN</td><td>Genscript/Desplan lab</td><td>-</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Guinea Pig anti-Traffic jam</td><td>Gift from D. Godt</td><td>-</td><td>Dilution: 1:2.000</td></tr><tr><td>Antibody: Donkey anti-Rat 405</td><td>Jackson ImmunoResearch</td><td>Code # 712-475-150; RRID: AB_2340680</td><td>Dilution: 1:100</td></tr><tr><td>Antibody: Donkey anti-Chicken 488</td><td>Jackson ImmunoResearch</td><td>Code #703-545-155; RRID: AB_2340375</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Donkey anti-Mouse 488</td><td>Jackson ImmunoResearch</td><td>Code #715-545-151; RRID: AB_2341099</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Donkey anti-Rabbit 488</td><td>Jackson ImmunoResearch</td><td>Code #711-545-152; RRID: AB_2313584</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Donkey anti-Rabbit A555</td><td>Thermo Scientific</td><td>Catalog #A-31572; RRID: AB_162543</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Donkey anti-Rat Cy3</td><td>Jackson ImmunoResearch</td><td>Code #712-165-153; RRID: AB_2340667</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Donkey anti-Guinea Pig 647</td><td>Jackson ImmunoResearch</td><td>Code #706-605-148; RRID: AB_2340476</td><td>Dilution: 1:200</td></tr><tr><td>Antibody: Donkey anti-Rat 647</td><td>Jackson ImmunoResearch</td><td>Code #712-605-153; RRID: AB_2340694</td><td>Dilution: 1:200</td></tr><tr><td>Antifade mountant</td><td>Thermo Fisher Scientific</td><td>S36940 (SlowFade Gold Antifade Mountant)</td><td>Other slowfades will work as well, including ones with DAPI (S36936)</td></tr><tr><td>Confocal Microscope</td><td>Zeiss</td><td>LSM-880</td><td>NA</td></tr><tr><td>Cover slips (22 x 30)</td><td>VWR</td><td>16004-332</td><td>NA</td></tr><tr><td>Fly: D. melanogaster: ab-GFP-FLAG</td><td>BDSC</td><td>38626; RRID: BDSC_38626</td><td>NA</td></tr><tr><td>Fly: D. melanogaster: Canton S</td><td>BDSC</td><td>64349; RRID: BDSC_64349</td><td>NA</td></tr><tr><td>Fly: D. melanogaster: dpp-Gal4</td><td>BDSC</td><td>1553; RRID: BDSC_1553</td><td>NA</td></tr><tr><td>Fly: D. melanogaster: ham-GFP.FPTB</td><td>BDSC</td><td>83660; RRID: BDSC_83660</td><td>NA</td></tr><tr><td>Fly: D. melanogaster: yw; 10xUAS-myr-GFP;</td><td>BDSC</td><td>32198; RRID: BDSC_32198</td><td>NA</td></tr><tr><td>Fly: D. melanogaster: UAS-Stinger</td><td>BDSC</td><td>84277; RRID: BDSC_84277</td><td>NA</td></tr><tr><td>Fly: D. melanogaster:&nbsp; wit-KO-Gal4</td><td>BDSC</td><td>600136; RRID: BDSC_600136</td><td>NA</td></tr><tr><td>Forceps</td><td>Dumont</td><td>#5 or #55s</td><td>NA</td></tr><tr><td>Glass slides</td><td>VWR</td><td>16005-106</td><td>NA</td></tr><tr><td>Glycine</td><td>Millipore Sigma</td><td>G7126-10MG</td><td>NA</td></tr><tr><td>HCR Amplification buffer</td><td>Molecular Instruments</td><td>HCR Gold RNA-FISH Kit</td><td>NA</td></tr><tr><td>HCR-Gold Amplifiers</td><td>Molecular Instruments</td><td>HCR Gold RNA-FISH Kit</td><td>NA</td></tr><tr><td>HCR HiFi Probes</td><td>Molecular Instruments</td><td>HCR Gold RNA-FISH Kit</td><td>NA</td></tr><tr><td>HCR Probe Hybridization Buffer</td><td>Molecular Instruments</td><td>HCR Gold RNA-FISH Kit</td><td>Contains formamide</td></tr><tr><td>HCR Probe Wash Buffer</td><td>Molecular Instruments</td><td>HCR Gold RNA-FISH Kit</td><td>Contains formamide</td></tr><tr><td>Nail polish (clear)</td><td>Electron Microscopy Sciences</td><td>72180</td><td>NA</td></tr><tr><td>Nuclease-free water (not DEPC-treated)</td><td>Thermo Fisher Scientific</td><td>AM9937</td><td>NA</td></tr><tr><td>Nutating mixer</td><td>Corning/Fisher Scientific</td><td>10-320-100</td><td>NA</td></tr><tr><td>Paraformaldehyde, 16% (w/v)</td><td>Electron Microscopy Sciences</td><td>15710</td><td>Toxic</td></tr><tr><td>PCR Machine/Thermal Cycler</td><td>Bio-Rad</td><td>1851148 (C1000 Touch Thermal Cycler)</td><td>NA</td></tr><tr><td>PCR pull-apart tube strips with dome cap strips (0.2 mL)&nbsp;</td><td>USA Scientific</td><td>#1402-2400</td><td>NA</td></tr><tr><td>Petri dishes (150 x 15 mm; for making dissecting dishes)</td><td>Fisher Scientific</td><td>FB0875714</td><td>NA</td></tr><tr><td>RNase ZAP</td><td>ThermoFisher Scientific</td><td>AM9780</td><td>NA</td></tr><tr><td>Slide spacer stickers</td><td>SUNJin</td><td>IS016 (iSpacer stickers (0,2 mm deep))</td><td>NA</td></tr><tr><td>Sylgard elastomer kit (for making dissecting dishes)</td><td>Fisher Scientific</td><td>NC9285739</td><td>NA</td></tr><tr><td>Triton X-100</td><td>Merck Millipore Sigma</td><td>X100</td><td>NA</td></tr></tbody></table>

fish de reação em cadeia de hibridização no lobo óptico em desenvolvimento de drosophila

Hybridization Chain Reaction FISH in the Developing Drosophila Optic Lobe

FISH de reação em cadeia de hibridização no lobo óptico em desenvolvimento de Drosophila

Understanding neural development requires precise characterization of when and where genes are expressed. Techniques such as in situ hybridization have ...

hybridization-chain-reaction-fish-developing-drosophila-optic

Research

JoVE Journal

Neuroscience

Neurociência

Usando Monte Whole In situ Hibridação a ligação de Biologia Molecular e Organísmicos

A arborização dendrítica de neurônios sensoriais...

Análise morfológica de Drosophila Larval sensorial periférica Dendrites Neuron e axônios Usando Mosaicos Genéticos

Todo o Monte RNA Fluorescente<em&gt; In situ</em&gt; A hibridação de<em&gt; Drosophila</em&gt; Embriões

O peixe-zebra, um pequeno peixe tropical, tornou-se um modelo popular para estudar a função do gene durante o desenvolvimento de vertebrados e doença. A expressão espacial e temporal dos genes alvo pode ser determinada...

Alta resolução Monte Whole<em&gt; In Situ</em&gt; Hibridização dentro embriões Zebrafish para estudar a expressão gênica e função

A imunocoloração é uma técnica eficaz para visualizar tipos específicos de células e proteínas dentro dos tecidos. Ao utilizar a sonicação, o protocolo descrito aqui alivia a necessidade de dissecar tecidos de Drosophila melanogaster de embriões e larvas em estágio avançado antes da imunocoloração. Fornecemos uma metodologia eficiente para a imunocoloração de larvas de montagem inteira fixadas em formaldeído.

Facilitou-Sonication Imunofluorescência Coloração de Late-estágio embrionário e larval<em&gt; Drosophila</em&gt; Tecidos<em&gt; In Situ</em

Nós descrevemos um monte de todo o processo de hibridação in situ multi-alvo cromogénico (MC-DESEJO) em embriões de Drosophila intactas permitindo a detecção simultânea e específica de três padrões de distribuição mRNA diferentes, contrastando precipitados de cor.

Multi-alvo Chromogenic Whole-mount<em&gt; In Situ</em&gt; Hibridização para Comparando Expressão Gênica em Domínios<em&gt; Drosophila</em&gt; Os embriões

O protocolo descrito de RNA em situ hibridização permite a detecção de RNA em toda Drosophila embriões ou tecidos dissecados. Usando placas de 96 poços da microplaca e amplificação do sinal de tyramide, transcrições podem ser detectadas no throughput, sensibilidade e alta resolução e a um custo relativamente baixo.

Alta resolução hibridação fluorescente In Situ em embriões da drosófila e tecidos usando amplificação de sinal Tyramide

Este manuscrito apresenta um método detalhado para a geração X-cromossomo braço sondas e realizando fluorescência situ hibridação in (peixe) para examinar o estado da irmã coesão cromátide em prometafase e metáfase prendi  Drosófila ovócitos. Este protocolo é apropriado para determinar se a coesão de braço meiótica é intacta ou interrompidos em diferentes genótipos.

Uso de hibridização in situ de fluorescência (FISH) para monitorar o estado da coesão do braço em oócitos de Drosophila na prometáfase e na metáfase I

O artigo descreve os métodos e reagentes necessários para realizar a hibridização em cadeia de reação em cadeia de hibridização por fluorescência completa de RNA (HCR, RNA, WM-FISH) para revelar informações sobre a resolução espacial e celular de genes de receptores quimiossensoriais na antena do mosquito e palpo maxilar.

Hibridização da Reação em Cadeia de RNA Fluorescência de Monte Inteiro Hibridização in situ de Genes Quimiossensoriais em Apêndices Olfativos de Mosquitos

A hibridização in situ de fluorescência iterativa assistida por expansão (EASI-FISH) é uma técnica robusta que combina microscopia de expansão com hibridização in situ de fluorescência (FISH) para detectar a expressão de vários genes em tecidos espessos. Este protocolo descreve os recentes avanços no método e sua adaptação para rotular o sistema nervoso central intacto da Drosophila.

Detecção multiplex da expressão gênica no cérebro intacto de Drosophila usando hibridização in situ de fluorescência iterativa assistida por expansão

New York University.  Este protocolo descreve as etapas necessárias para realizar hibridização fluorescente in situ por reação em cadeia (HCR-FISH) nos lobos ópticos larvais de Drosophila melanogaster . Procedimentos para o projeto da sonda, hibridização, amplificação e montagem de tecidos são detalhados para facilitar a visualização do lobo óptico em desenvolvimento.

FISH de reação em cadeia de hibridização no lobo óptico em desenvolvimento de Drosophila

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