Summary
笼状化合物的光解允许快速和本地化的各种生理活性化合物的浓度增加生产。在这里，我们将展示如何获得光解笼中的cAMP结合或分离小鼠嗅感觉神经元的嗅觉传导的研究为笼钙膜片钳记录。
Abstract
笼状化合物的光解允许快速和本地化的各种生理活性化合物的浓度增加生产。笼中的化合物是由一种化学笼紫外线闪光灯可以打破生理不活跃分子。在这里，我们展示了如何获取结合分离小鼠嗅感觉神经元的嗅觉传导的研究笼化合物光解的膜片钳记录。嗅觉传导的过程（图1）在嗅感觉神经元，气味结合的受体导致cAMP的增加，打开环核苷酸门控（CNG）， 通道 2的纤毛。 CA通过CNG的渠道进入激活钙激活Cl通道。我们展示了如何脱离鼠标嗅上皮和如何CNG通道或钙激活的氯通道激活，光解笼中的 cAMP 4笼约5元</ SUP>。我们使用闪光灯6,7申请紫外线闪烁睫状肌地区uncage阵营或Ca，而膜片钳记录采取措施，目前在全细胞电压钳配置 8-11 。
Discussion
闪存与膜片钳记录相结合的笼化合物的光解是一种有用的技术，获得快速和地方在内外细胞的生理活性分子的浓度跳跃。合成了几种类型的笼compounds1，这种技术可以适用于各种类型的细胞，包括培养细胞中表达，可激活或由一些可用的笼化合物11的光解调制的离子通道。
笼状化合物的光解需要高强度的近紫外线光脉冲uncage在很短的时间足够数量的分子。可用于各种光源：一个不断汞或氙弧灯控制快门，再加啶端口显微镜，氙气闪光灯，紫外激光，和最近开发的高功率紫外线灯发光二极管（LED ）。每个类型的光源优势和disadvan产品关键词根据具体应用和设备的成本。一个闪光灯相比，连续运行的灯有一个较低的光照强度，因此快门控制的光脉冲的持续时间，需要增加多达数百毫秒uncaged分子获得足够数量。紫外激光是非常昂贵的。高功率紫外发光二极管闪光光解14最近市售其他方法可以提供一个很好的选择。然而，闪光灯的优势是，他们有一个更广泛的发射光谱比紫外发光二极管，允许使用uncaging在我们的应用程序使用一个氙气闪光灯的主要优点是与不同的光谱特征笼化合物的几种类型：一个好的时间分辨率，确实光脉冲的持续时间约1毫秒;一个广泛的紫外光谱，是适合光化学性质不同的分子光解的可能性选择毛钱nsion的光点照亮睫状肌的地区，可以方便地选择不同的光照强度 6 。此外，氙气闪光灯的灯有一个合理的成本，这是很容易实现电设置，不需要特殊的维护。
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Disclosures
没有利益冲突的声明。
Materials
| Name | Company | Catalog Number | Comments |
| Adapter module flash lamp to microscope | Rapp OptoElectronic | FlashCube 70 | |
| Air table | TMC | MICRO-g 63-534 | |
| Digitizer | Axon Instruments | Digidata 1322A | |
| Data Acquisition Software | Axon Instruments | pClamp 8 | |
| Data Analysis Software | WaveMetrics | Igor | |
| Mirror for adapter module | Rapp OptoElectronic | M70/100 | |
| Electrode holder | Axon Instruments | 1-HL-U | |
| Faraday’s cage | Custom Made | ||
| Filter cube | Olympus Corporation | U-MWU | Excitation filter removed |
| Flash lamp | Rapp OptoElectronic | JML-C2 | |
| Forceps Dumont #55 | World Precision Instruments, Inc. | 14099 | |
| Glass capillaries | World Precision Instruments, Inc. | PG10165-4 | |
| Glass bottom dish | World Precision Instruments, Inc. | FD35-100 | |
| Illuminator | Olympus Corporation | Highlight 3100 | |
| Inverted microscope | Olympus Corporation | IX70 | |
| Micromanipulators | Luigs & Neumann | SM I | |
| Micropipette Puller | Narishige International | PP-830 | |
| Monitor | HesaVision | MTB-01 | |
| Neutral density filters | Omega Optical | varies | |
| Objective 100X | Carl Zeiss, Inc. | Fluar 440285 | Either Zeiss or Olympus |
| Objective 100X | Olympus Corporation | UPLFLN 100XOI2 | Either Zeiss or Olympus |
| Optical UV shortpass filter | Rapp OptoElectronic | SP400 | |
| Patch-clamp amplifier | Axon Instruments | Axopatch 200B | |
| Photo Diode Assembly | Rapp OptoElectronic | PDA | |
| Quartz light guide | Rapp OptoElectronic | varies | We use 600 μm diameter |
| Silver wire | World Precision Instruments, Inc. | AGT1025 | |
| Silver ground pellet | Warner Instruments | 64-1309 | |
| Xenon arc lamp | Rapp OptoElectronic | XBL-JML | |
| Reagent | Company | Catalogue number | |
| BCMCM-caged cAMP | BioLog | B016 | |
| Bovine serum albumin (BSA) | Sigma-Aldrich | A8806 | |
| CaCl2 standard solution 0.1 M | Fluka | 21059 | |
| Caged Ca: DMNP-EDTA | Invitrogen | D6814 | |
| Cysteine | Sigma-Aldrich | C9768 | |
| Concanavalin A type V (ConA) | Sigma-Aldrich | C7275 | |
| CsCl | Sigma-Aldrich | C4036 | |
| DMSO | Sigma-Aldrich | D8418 | |
| DNAse I | Sigma-Aldrich | D4527 | |
| EDTA | Sigma-Aldrich | E9884 | |
| EGTA | Sigma-Aldrich | E4378 | |
| Glucose | Sigma-Aldrich | G5767 | |
| HEPES | Sigma-Aldrich | H3375 | |
| KCl | Sigma-Aldrich | P3911 | |
| KOH | Sigma-Aldrich | P1767 | |
| Leupeptin | Sigma-Aldrich | L0649 | |
| MgCl2 | Fluka | 63020 | |
| Papain | Sigma-Aldrich | P3125 | |
| Poly-L-lysine | Sigma-Aldrich | P1274 | |
| NaCl | Sigma-Aldrich | S9888 | |
| NaOH | Sigma-Aldrich | S5881 | |
| NaPyruvate | Sigma-Aldrich | P2256 |
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