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<ol>
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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>Verdunning: 1:20</td></tr><tr><td>Antibody: Chicken anti-GFP</td><td>Millipore Sigma</td><td>Catalog# 06-896; RRID: AB-619712</td><td>Verdunning: 1:500</td></tr><tr><td>Antibody: Rabbit anti-GFP</td><td>Life Technologies</td><td>#11122; RRID: AB_221569</td><td>Verdunning: 1:500</td></tr><tr><td>Antibody: Rabbit anti-Dll</td><td>Genscript/Desplan lab</td><td>-</td><td>Verdunning: 1:200</td></tr><tr><td>Antibody: Rat anti-NCadherin</td><td>DSHB</td><td>DN-Ex #8, RRID: AB_528121</td><td>Verdunning: 1:50</td></tr><tr><td>Antibody: Rabbit anti-SoxN</td><td>Genscript/Desplan lab</td><td>-</td><td>Verdunning: 1:200</td></tr><tr><td>Antibody: Guinea Pig anti-Traffic jam</td><td>Gift from D. Godt</td><td>-</td><td>Verdunning: 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>Verdunning: 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>Verdunning: 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>Verdunning: 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>Verdunning: 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>Verdunning: 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>Verdunning: 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>Verdunning: 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>Verdunning: 1:200</td></tr><tr><td>Antifade mountant</td><td>Thermo Fisher Scientific</td><td>S36940 (SlowFade Gold Antifade Mountant)</td><td>Andere slowfades zullen ook werken, inclusief degenen met 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 of #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>Bevat formamide</td></tr><tr><td>HCR Probe Wash Buffer</td><td>Molecular Instruments</td><td>HCR Gold RNA-FISH Kit</td><td>Bevat 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>Toxisch</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>

hybridisatiekettingreactie fish in de zich ontwikkelende drosophila optische lob

Hybridization Chain Reaction FISH in the Developing Drosophila Optic Lobe

Hybridisatiekettingreactie FISH in de zich ontwikkelende Drosophila optische lob

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

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Neuroscience

Neurowetenschap

Met behulp van Whole Mount In situ Hybridisatie aan Moleculaire Biologie en organismale Link

De dendritische arborization sensorische neuronen van...

Morfologische Analyse van de Drosophila Larvale Perifere sensorische neuron dendrieten en axonen het gebruik van genetische Mozaïeken

Hier beschrijven we een geheel gemonteerde...

Hele Mount RNA TL<em&gt; In situ</em&gt; Hybridisatie van<em&gt; Drosophila</em&gt; Embryo&#39;s

De zebravis, een kleine tropische vissen, is uitgegroeid tot een populair model voor het bestuderen van gen-functie tijdens gewervelde ontwikkeling en ziekte. De ruimtelijke en temporele expressie van de doelwitgenen worden...

Hoge resolutie Hele Mount<em&gt; In Situ</em&gt; Hybridisatie in zebravis embryo&#39;s om genexpressie en functie te bestuderen

Immuunkleuring is een effectieve techniek voor het visualiseren van specifieke celtypen en eiwitten in weefsels. Door het gebruik van sonicatie vermindert het hier beschreven protocol de noodzaak om Drosophila melanogaster weefsels van laat-stadium embryo's en larven te dissekeren voordat immunokleuring wordt uitgevoerd. We bieden een efficiënte methodologie voor de immunokleuring van formaldehyde-gefixeerde hele larven.

-Sonicatie vergemakkelijkt immunofluorescentiekleuring van Late-fase embryonale en larvale<em&gt; Drosophila</em&gt; Weefsels<em&gt; In Situ</em

We beschrijven een multi-target chromogene whole-mount in situ hybridisatie (MC-WISH) procedure in intacte Drosophila embryo's toestaan ​​gelijktijdige en specifieke detectie van drie verschillende mRNA distributie patronen door contrasterende kleur neerslagen.

Multi-target Chromogene Whole-mount<em&gt; In Situ</em&gt; Hybridisatie voor het vergelijken van genexpressie Domeinen in<em&gt; Drosophila</em&gt; Embryo

Het beschreven RNA in situ hybridisatie protocol staat toe de opsporing van RNA in hele Drosophila embryo's of ontleed weefsels. Met behulp van 96-wells microtiterplaat platen en tyramide signaal versterking, kunnen afschriften worden gedetecteerd op hoge resolutie, gevoeligheid en doorvoer, en tegen een relatief lage prijs.

Hoge resolutie fluorescentie In Situ hybridisatie in Drosophila embryo's en weefsels met behulp van Tyramide signaal versterking

Dit manuscript presenteert een gedetailleerde methode voor het genereren van X-chromosoom arm sondes en presterende fluorescentie in situ hybridisatie (FISH) te onderzoeken van de Braziliaanse deelstaat zus chromatide cohesie in prometaphase en metafase ik gearresteerd  Drosophila eicellen. Dit protocol is geschikt om te bepalen of cohesie Meiotische arm intact of verstoord in verschillende genotypen is.

Met behulp van fluorescentie In Situ hybridisatie (FISH) te controleren van de toestand van Arm cohesie in Prometaphase en metafase ik Drosophila eicellen

Het artikel beschrijft de methoden en reagentia die nodig zijn om hybridisatiekettingreactie-RNA, whole-mount fluorescentie in situ hybridisatie (HCR, RNA, WM-FISH) uit te voeren om inzicht te krijgen in de ruimtelijke en cellulaire resolutie van chemosensorische receptorgenen in de muggenantenne en maxillaire palp.

Hybridisatie Kettingreactie RNA Whole-Mount Fluorescentie In situ Hybridisatie van chemosensorische genen in olfactorische aanhangsels van muggen

Expansie-ondersteunde iteratieve fluorescentie in situ hybridisatie (EASI-FISH) is een robuuste techniek die expansiemicroscopie combineert met fluorescentie in situ hybridisatie (FISH) om de expressie van meerdere genen in dik weefsel te detecteren. Dit protocol schetst recente vorderingen in de methode en de aanpassing ervan voor het labelen van het intacte Drosophila centrale zenuwstelsel..

Multiplex detectie van genexpressie in de intacte Drosophila hersenen met behulp van expansie-ondersteunde iteratieve fluorescentie in situ hybridisatie

New York University.  Dit protocol beschrijft de stappen die nodig zijn om hybridisatiekettingreactie fluorescerende in situ hybridisatie (HCR-FISH) uit te voeren in oogtische lobben van Drosophila melanogaster-larven . Procedures voor probe-ontwerp, hybridisatie, amplificatie en weefselmontage worden gedetailleerd om de visualisatie van de zich ontwikkelende optische kwab te vergemakkelijken.

Hybridisatiekettingreactie FISH in de zich ontwikkelende Drosophila optische lob

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