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R-Scripts and Fiji-macro used for the construction of kymographs based on fluorescence profiles

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GiacomoGiacomelli/Cell-Profiles-and-Demographs

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Cell profiles and demographs

Available Files:

Kymographs_from_Cell_Profiles_V3.pptx -> This .pptx file contains extensive explanation on how to use the various scripts/macros contained within this page

ProfilingCells_EPI.ijm

  • Input: Multi Channel Microscopy Image (Any Image)
  • Input: RoiSet.zip (Segmented Lines with defined Width -> User Defined)
  • Output1: "Gray*.txt" (Position X, Channel 1 Intensity from ROI #*)
  • Output2: "Red*.txt" (Position X, Channel 2 Intensity from ROI #*)
  • Output3: "Blue*.txt" (Position X, Channel 3 Intensity from ROI #*)
  • Extra requirements 1: Create a new folder named "Profiles" in the same folder containing the image
  • Extra requirements 2: Rename the folder after completing an image "Profiles" -> "ProfilesN" (Repeat "Extra requirements 1" if analysing a second image)

CPAD1_Extract_Cell_length_from_membrane_stain_V1.R

  • Input: "Profiles*" folders containg fluorescence profiles of membrane stained cells (Usually "Red*.txt")
  • Ouput: Curated Profiles ("Gray*.txt","Red*.txt","Blue*.txt")
  • Extra requirements: Create a "Membranes*" with the same numbering of the "Profiles*" folders

CPAD2_Cell_Profiles_And_Demographs_V3.R NOTE: This Script uses the "findpeaks" function from -> https://rdrr.io/bioc/alsace/man/fitpeaks.html

  • Input1: "Membranes*" or "Profiles*" folders containing fluorescence profiles ("Gray*.txt", "Red*.txt, "Blue*.txt").
  • User Defined1: PlotType ("CellNorm/PopNorm") -> determines the type of normalization
  • User Defined2: m100/mayim OR 1+(m_c100/mayim_c) -> determines the type of normalization
  • Output1: "../Profiles_ordered.txt" -> file containing the fluorescence profiles ordered by cell length
  • Output2: "../Profiles_ordered_matrix.txt" -> matrix used to represent the fluorescence profiles as demographs via hist2d()
  • Output3: "../maxim.txt" -> maximum length among the profiles
  • Output4: "../Demograph.png" -> demograph
  • Output5: "../Cell*.png" -> Fluorescence profiles and peak position
  • Output6: "../CellLength_and_Peaks.txt" -> file containing cell length and the number of peaks contained in each fluorescence channel

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R-Scripts and Fiji-macro used for the construction of kymographs based on fluorescence profiles

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