RICS Simulations - Cell Migration GatewayRICS模拟细胞迁移的网关.pptVIP

RICS Simulationsfor two photon excitation Opening page of GLOBALS for Images program: Click on the SimFCS buttom First page of SimFCS: Under ACTIONS in the first page of SIMFCS Go to “simulate FCS data” Set up Point Spread Function (PSF) Click on Particles: Select Number of molecules, brightness and shape (1-4) Exclude: Misc., Stick/Disk, and Reaction Click on Time/Scan Now you are ready to begin the simulations. Click on START SIMULATION Save your Simulated Data RICS: Fitting the ACF For additional simulations: Close fit and plot of fitted spatial correlation Close RICS Back to SimFCS first page Click on Simulate data Simulation: D=5?m2/s Simulation: 20?m2/s  Simulation: D=100?m2/s(not confined) Special conditions: * * Click on Parameter Pixel size In the PSF page you can select the shape of the point spread function. The 3-D Gaussian has independent radial and axial waist, while for the Gaussian-Lorentzian, the z-extension of the function depends on the radial waist. Also, the 3-D Gaussian is not squared to provide intensity while the Gaussian-Lorentzian is squared. Radial and axial waist have micron units. The radial waist for x and y should be the same (~0.25-0.55) and the axial waist is five times the radial waist. The box size parameters determine the half-size of the box used for the simulation. The conversion factor between physical size and granularity of the box is that each unit of the box corresponds to 50 nm. Generally, this is enough for most optical instruments. A box size of 128 is a good starting point. When you have set up PSF click on particles button Is fixed Clock frequency determines the maximum Diffusion allowed Use the particle editor page to select properties of the particles for simulation. The are a maximum of 50 columns, which represents 50 different types of molecules. The total number of molecules (the sum of all species) cannot exceed 4000. The first column determines how many molecules of a given property are in