PATCHMASTER TUTORIAL

Contents1 Fluorescence Ratio Measurements 11.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.2 Configuring Patchmaster for Fluorescence Measurements 21.2.1 Turning on the Photometry Extension . . . . . . . 31.3 Editing the PulseGenerator Sequences . . . . . . . . . . . . 31.3.1 The Sequence ”TestFura” . . . . . . . . . . . . . . . 31.3.2 The ”RatioFura” . . . . . . . . . . . . . . 61.4 Editing the Online Analysis . . . . . . . . . . . . . . . . . . 61.4.1 The ”Background” Analysis Method . . . . . . . . . 71.4.2 The ”Ratio” Analysis Method . . . . . . . . . . . . . 81.5 The Experiment with Photometry Measurements . . . . . . 92 High-Speed Fluorescence Measurements 112.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . 112.2 Editing the pgf-File. . . . . . . . . . . . . . . . . . . . . . . 112.2.1 The Photometry Stimulation . . . . . . . . . . . . . 132.2.2 Extraction of the F340 and F380 Signals . . . . . . . 152.2.3 Calculation of the Ratio Trace . . . . . . . . . . . . 192.3 Display of the Data in the Oscilloscope window . . . . . . . 192.4 Calculation of Compression Settings . . . . . . . . . . . . . 201. Fluorescence RatioMeasurements1.1 IntroductionInthefollowingtutorialwewilldescribeawhole-cellexperimentwithload-ing a ratiometric fluorescence dye (e.g. fura-2) through the patch pipetteinto the cell.Beforewestartthedetaileddescriptionhowtoset-upandconfigurePatch-master, we will first outline the ...