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localfsc [2015/09/04 12:12]
gcardone created
localfsc [2019/06/19 16:22] (current)
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 LocalFSC is a plugin for Chimera to estimate the resolution in selected subregions of a EM map. LocalFSC is a plugin for Chimera to estimate the resolution in selected subregions of a EM map.
  
-The resolution is estimated by means of the Fourier Shell Correlation (FSC), calculated from two independent maps. Generally these are the maps each generated from half the particles in a data set. Selection is performed by placing markers in the regions of interest, and the size of the regions to analyze ​can be set either manually or automatically. Comments can be added to each measure, and all the estimates can be saved in a text file for further analysis.+The resolution is estimated by means of the Fourier Shell Correlation (FSC), calculated from two independent maps. Generally these are the maps each generated from half the particles in a data set. Selection is performed by placing markers in the regions of interest, and the size of the regions to analyse ​can be set either manually or automatically. Comments can be added to each measure, and all the estimates can be saved in a text file for further analysis. 
 + 
 +<WRAP center round info 75%> 
 +Local resolution analysis of the entire map can be done, using the same methodological approach, with programs incorporated into [[http://​lsbr.niams.nih.gov/​bsoft/​|Bsoft]]. Specifically,​ local analysis and local adaptive filtering can be performed with //blocres// and //​blocfilt//,​ respectively. For further information please read the article cited below. 
 +</​WRAP>​ 
 + 
  
 ===== Installation ===== ===== Installation =====
  
-  *     Create ​new directory that will contain the plugin+  *     If you don't already have a directory ​on your computer ​that is specific for external plugins for chimera, create one
-  *     ​Download ​and unzip the file [[http://cryoem.ucsd.edu/​programs/​code/​LocalFSC_chimera.tar.gz|LocalFSC_chimera.tar.gz]] in this new directory. +  *     ​Download the file [[http://bakerlab.ucsd.edu/​programs/​code/​LocalFSC_chimera.tar.gz|LocalFSC_chimera.tar.gz]] in this directory ​and uncompress it
-  *     Open Chimera ​and go to Favorites/Preferences+  *     In Chimerago to Favorites->Preferences, from Category select ​Tools and add the directory containing the LocalFSC folder to the Locations.
-  *     ​Select "Tools" Category ​and add the directory containing the LocalFSC folder to the "Locations".+
  
-The LocalFSC module ​now should appear ​under Tools->​Volume Data menu. +The LocalFSC module ​will be under the Tools->​Volume Data menu. 
  
  
  
 ===== Usage ===== ===== Usage =====
-A test data set is provided together with tutorial which describes ​the procedure ​to get from the raw micrographs to a helical reconstruction. +Once opened, the LocalFSC module, ​help button provides all the basic information on how to use it and on all the different options available in the interface.
- +
-[[http://​cryoem.ucsd.edu/​programs/​IHRSR++tutorial_v1.5.pdf|Tutorial]] (pdf)+
  
-[[http://cryoem.ucsd.edu/​programs/​code/​IHRSR_test_prob.tar.gz|Test Dataset]] (**~5 GB file**) +In order to get familiar with the module, here is provided a [[http://bakerlab.ucsd.edu/​programs/​code/​TestCaseLFSC.tar.gz|Test Dataset]] ​to download, which contains a simulated reconstruction of a ribosome ​(see article below for details). Specifically,​ the archive file contains three maps (two half maps and one map from all the particles) and a Chimera marker file with two markers pointing to two domains with different occupancy level. 
--------------------------+  ​Uncompress the data. 
 +  ​Open the tree maps and the marker ​file in Chimera. 
 +  ​Verify that all three maps are overlapping and their pixel size is properly set in the Features->​Coordinates menu of the Volume Viewer dialog window. 
 +  ​Hide the two half maps. 
 +  * Open the module (Tools->Volume Data->​LocalFSC in the main window or Tools->​LocalFSC in Volume Viewer). 
 +  * In Settings, point the first and second map to the two half maps. 
 +  * Still in Settings, set the maximum resolution available (i.e. unfiltered out) in the half maps (in this case 10). 
 +  * In the main window, select one of the markers with Ctrl-left click and the press the Estimate button in Resolution. The estimate will be listed in the GUI, and the FSC plot will be shown in a separate window. 
 +  * You can select multiple markers where to estimate the resolution. In this case the program will not show their corresponding FSC plots. ​
  
 ===== Credits ===== ===== Credits =====