1. Instrumentation 
<ol>
<li>To measure the response of olfactory sensory neurons to photolysis of caged compounds we use a flash lamp in combination with a typical patch-clamp recording system including: a patch-clamp amplifier, a recording electrode and a reference electrode connected to the head-stage of a patch-clamp amplifier, a digitizer, a computer, software for data acquisition, micro-manipulators, an epifluorescence microscope, a perfusion system, an anti-vibration table and a Faraday cage (Figure 2).</li>
<li...

<ol>
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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flash+photolysis+of+caged+compounds.">Flash photolysis of caged compounds.</a> Microelectrodes: Theory and Applications. , (1991).</li><li>Lagostena, L., Menini, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-cell+recordings+and+photolysis+of+caged+compounds+in+olfactory+sensory+neurons+isolated+from+the+mouse.">Whole-cell recordings and photolysis of caged compounds in olfactory sensory neurons isolated from the mouse.</a> Chem. 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Flash fotolyse i bur forbindelser kombineret med patch-clamp optagelser er en teknik til at opnå hurtige og lokale hopper i koncentrationen af ​​fysiologisk aktive molekyler både inden for og uden for cellerne. Flere typer af bur compounds1 er blevet syntetiseret, og denne teknik kan anvendes til forskellige typer af celler, herunder dyrkede celler, der udtrykker ion-kanaler, der kan aktiveres eller afpasses efter fotolyse af nogle af de tilgængelige bur forbindelser 11. Fo...

<table border="1"><tbody><tr><th> Udstyr </th><th> Firma </th><th> Katalog nummer </th><th> Kommentarer </th></tr><tr><td> Adaptermodul flash lampe til mikroskop </td><td> Rapp optoelektroniske </td><td> FlashCube 70 </td><td></td></tr><tr><td> Air tabel </td><td> TMC </td><td> MICRO-g 63-534 </td><td></td></tr><tr><td> Digitizer </td><td> Axon Instrumenter </td><td> Digidata 1322A </td><td></td></tr><tr><td> Data Acquisition Software </td><td> Axon Instrumenter </td><td> pClamp 8 </td><td></td></tr><tr><td> Data Analysis Software </td><td> WaveMetrics </td><td> Igor </td><td></td></tr><tr><td> Spejl for adaptermodul </td><td> Rapp optoelektroniske </td><td> M70/100 </td><td></td></tr><tr><td> Elektrode holder </td><td> Axon Instrumenter </td><td> 1-HL-U </td><td></td></tr><tr><td> Faradays bur </td><td> Skræddersyet </td><td></td><td></td></tr><tr><td> Filter terning </td><td> Olympus </td><td> U-MAE </td><td> Excitation filter fjernes </td></tr><tr><td> Flash lampe </td><td> Rapp optoelektroniske </td><td> JML-C2 </td><td></td></tr><tr><td> Tang Dumont # 55 </td><td> Verdens præcisionsinstrumenter </td><td> 14099 </td><td></td></tr><tr><td> Glas kapillærer </td><td> Verdens præcisionsinstrumenter </td><td> PG10165-4 </td><td></td></tr><tr><td> Glas nederste skaal </td><td> Verdens præcisionsinstrumenter </td><td> FD35-100 </td><td></td></tr><tr><td> Illuminator </td><td> Olympus </td><td> Fremhæv 3100 </td><td></td></tr><tr><td> Inverteret mikroskop </td><td> Olympus </td><td> IX70 </td><td></td></tr><tr><td> Micromanipulators </td><td> Luigs &amp; Neumann </td><td> SM Jeg </td><td></td></tr><tr><td> Mikropipette Puller </td><td> Narishige </td><td> PP-830 </td><td></td></tr><tr><td> Monitor </td><td> HesaVision </td><td> MTB-01 </td><td></td></tr><tr><td> Neutralfiltre </td><td> Omega Optical </td><td> varierer </td><td></td></tr><tr><td> Formål 100X </td><td> Zeiss </td><td> Fluar 440285 </td><td> Enten Zeiss eller Olympus </td></tr><tr><td> Formål 100X </td><td> Olympus </td><td> UPLFLN 100XOI2 </td><td> Enten Zeiss eller Olympus </td></tr><tr><td> Optisk UV shortpass filter </td><td> Rapp optoelektroniske </td><td> SP400 </td><td></td></tr><tr><td> Patch-clamp forstærker </td><td> Axon Instrumenter </td><td> Axopatch 200B </td><td></td></tr><tr><td> Foto Diode Assembly </td><td> Rapp optoelektroniske </td><td> PDA </td><td></td></tr><tr><td> Quartz lysleder </td><td> Rapp optoelektroniske </td><td> varierer </td><td> Vi bruger 600 μm diameter </td></tr><tr><td> Silver wire </td><td> Verdens præcisionsinstrumenter </td><td> AGT1025 </td><td></td></tr><tr><td> Silver jorden pellet </td><td> Warner instrumenter </td><td> 64-1309 </td><td></td></tr><tr><td> Xenon arc lampe </td><td> Rapp optoelektroniske </td><td> XBL-JML </td><td></td></tr></tbody></table> <table border="1"><tbody><tr><th> Reagens </th><th> Firma </th><th> Katalog nummer </th></tr><tr><td> BCMCM-bur cAMP </td><td> Biologist </td><td> B016 </td></tr><tr><td> Bovin serum albumin (BSA) </td><td> Sigma </td><td> A8806 </td></tr><tr><td> CaCl2 standardløsning 0,1 M </td><td> Fluka </td><td> 21059 </td></tr><tr><td> Caged Ca: DMNP-EDTA </td><td> Invitrogen </td><td> D6814 </td></tr><tr><td> Cystein </td><td> Sigma </td><td> C9768 </td></tr><tr><td> Concanavalin En type V (Cona) </td><td> Sigma </td><td> C7275 </td></tr><tr><td> CsCl </td><td> Sigma </td><td> C4036 </td></tr><tr><td> DMSO </td><td> Sigma </td><td> D8418 </td></tr><tr><td> DNAse Jeg </td><td> Sigma </td><td> D4527 </td></tr><tr><td> EDTA </td><td> Sigma </td><td> E9884 </td></tr><tr><td> EGTA </td><td> Sigma </td><td> E4378 </td></tr><tr><td> Glucose </td><td> Sigma </td><td> G5767 </td></tr><tr><td> HEPES </td><td> Sigma </td><td> H3375 </td></tr><tr><td> KCl </td><td> Sigma </td><td> P3911 </td></tr><tr><td> KOH </td><td> Sigma </td><td> P1767 </td></tr><tr><td> Leupeptin </td><td> Sigma </td><td> L0649 </td></tr><tr><td> MgCl2 </td><td> Fluka </td><td> 63020 </td></tr><tr><td> Papain </td><td> Sigma </td><td> P3125 </td></tr><tr><td> Poly-L-lysin </td><td> Sigma </td><td> P1274 </td></tr><tr><td> NaCl </td><td> Sigma </td><td> S9888 </td></tr><tr><td> NaOH </td><td> Sigma </td><td> S5881 </td></tr><tr><td> NaPyruvate </td><td> Sigma </td><td> P2256 </td></tr></tbody></table>

flash photolysis of caged compounds in the cilia of olfactory sensory  neurons

Fotolyse i bur forbindelser tillader produktion af hurtige og lokaliserede stigninger i koncentrationen af forskellige fysiologisk aktive stoffer 1. Caged forbindelser er molekyler lavet fysiologisk inaktiv ved en kemisk bur, der kan brydes af en flash af ultraviolet lys. Her viser vi hvordan du får patch-clamp optagelser kombineret med fotolyse i bur forbindelser til studiet af olfaktoriske transduktion i dissocierede mus lugtesansen sensoriske neuroner. Processen af olfaktoriske transduktion (Figur 1) finder sted i cilia af olfaktoriske sensoriske neuroner, hvor lugtstof binding til receptorer medfører en forøgelse af cAMP, som åbner cyklisk nukleotid-gated (CNG) kanaler 2. Ca indlæg via CNG kanaler aktiverer Ca-aktiverede Cl-kanaler. Vi viser hvordan man kan adskille neuroner fra musen lugtepithelet 3 og hvordan du aktiverer CNG kanaler eller Ca-aktiverede Cl kanaler ved fotolyse i bur cAMP 4 eller bur Ca 5 &lt;/ Sup&gt;. Vi bruger en flash lampe 6,7 til at anvende ultraviolet blinker til ciliaere region til uncage lejr eller Ca, mens patch-clamp optagelser er taget for at måle strøm i hel-celle spænding-clamp konfiguration 8-11.

Watch this Scientific Journal Video about Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory  Neurons at JoVE.com

Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory  Neurons

Fotolyse i bur forbindelser tillader produktion af hurtige og lokaliserede stigninger i koncentrationen af ​​forskellige fysiologisk aktive stoffer. Her viser vi hvordan du får patch-clamp optagelser kombineret med fotolyse i bur cAMP eller anbragt i bur Ca for studiet af olfaktoriske transduktion i dissocierede mus lugtesansen sensoriske neuroner.

Flash fotolyse af Caged forbindelser i Cilia af olfaktoriske sensoriske neuroner

Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds1. ...

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