R/baseGUI.R

In genArise: Microarray Analysis tool

# Windows for diagnostic plots and the analysis
require(tkrplot)
require(xtable)

back.gui <- function(envir){
  flag <- 1
  answer <- as.character(tkmessageBox(message = "All the performed operations will be deleted from this project. Are you sure?",
                                      icon = "question", type = "yesnocancel", default = "yes"))
  if( answer == "yes" ){
    proj.file <- read.table(get("history.project", envir = envir),header=FALSE,sep="\t")
    proj.tmp <- proj.file[1:(length(proj.file[,1])-get("op.counter",envir = envir)),]
    assign("op.counter",0,envir = envir)
    reset.history(get("history.project", envir = envir),proj.tmp)
    flag <- 0
  }
  flag
}

principal <-  function(envir,swap=0){
  tclRequire("BWidget")
  tt <- tktoplevel()
  tkfocus(tt)
  o.spot <- get("o.spot", envir = envir)
  datos <- attr(o.spot, "spotData")
  M <- log(datos$Cy5, 2) - log(datos$Cy3, 2)
  tkwm.geometry(tt,"+50+50")
  tkwm.title(tt,"GenArise Microarray Analyzer")
  
  lower.menu <- tkframe(tt, relief = "groove", borderwidth=2)
  
  image.menu.frame <- tkframe(lower.menu)
  
  tcl("image","create","photo","analysis",file=file.path(get("icons.dir", envir = envir),"icons/analysis.png"))
  analysis.button <- tkbutton(image.menu.frame,image="analysis", command = function(){ follow.wizard()})
  tkconfigure(analysis.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","acroread",file= file.path(get("icons.dir", envir = envir),"icons/acroread.png"))
  acroread.button <- tkbutton(image.menu.frame,image="acroread", command = function(){save.as.pdf()}) 
  tkconfigure(acroread.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","editor",file=file.path(get("icons.dir", envir = envir),"icons/editor.png"))
  editor.button <- tkbutton(image.menu.frame,image="editor", command = function() note(envir))
  tkconfigure(editor.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","back",file=file.path(get("icons.dir", envir = envir),"icons/regresar.png"))
  back.button <- tkbutton(image.menu.frame,image="back",command =  function(){
    genArise(); tkdestroy(tt)
    unlink(paste(get("path.project", envir = envir),".prj",sep=""))
    unlink(get("path.project", envir = envir), recursive = TRUE)})
  tkconfigure(back.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","help",file=file.path(get("icons.dir", envir = envir),"icons/help.png"))
  help.button <- tkbutton(image.menu.frame,image="help", command = function() help())
  tkconfigure(help.button, heigh = 16, width = 16)

  tcl("image","create","photo","exit",file=file.path(get("icons.dir", envir = envir),"icons/logout.png"))
  exit.button <- tkbutton(image.menu.frame,image="exit", command = function(){
    tkdestroy(tt)})
  tkconfigure(exit.button, heigh = 16, width = 16)
  
  tkgrid(analysis.button, acroread.button, editor.button, back.button, help.button, exit.button, sticky = "nw")
  
  etiqueta <- tklabel(lower.menu, text = "")
  tkconfigure(etiqueta, text = "", width = "90") 
  tkgrid(image.menu.frame, etiqueta, sticky = "w")
  tkgrid(lower.menu)
  
  label.function <-  function(texto.etiqueta) tkconfigure(etiqueta, text = texto.etiqueta, width = "90")
  
  
  tkbind(analysis.button, "<Enter>", function() label.function("Make analysis data"))
  tkbind(analysis.button, "<Leave>", function() label.function(""))
  
  tkbind(acroread.button, "<Enter>", function() label.function("Save graphic as PDF"))
  tkbind(acroread.button, "<Leave>", function() label.function(""))
  
  tkbind(editor.button, "<Enter>", function() label.function("Notes about this experiment"))
  tkbind(editor.button, "<Leave>", function() label.function(""))
  
  tkbind(help.button, "<Enter>", function() label.function("Display the genArise's help"))
  tkbind(help.button, "<Leave>", function() label.function(""))

  tkbind(exit.button, "<Enter>", function() label.function("Exit GenArise GUI"))
  tkbind(exit.button, "<Leave>", function() label.function(""))

  
  follow.wizard <- function(){
    respuesta <- as.character(tkmessageBox(message = "Do you want to follow the wizard?",
                                           icon = "question", type = "yesnocancel", default = "yes"))
    
    if(respuesta == "yes"){ 
      tkconfigure(tt,cursor="watch")
      tkentryconfigure(topMenu, "0", state ="disable" )
      tkentryconfigure(topMenu, "1", state ="disable" )
      tkentryconfigure(topMenu, "2", state ="disable" )
      tkentryconfigure(topMenu, "3", state ="disable" )
      tkentryconfigure(topMenu, "4", state ="disable" )
      analysis.window("", follow.wizard = TRUE, envir,swap)
      tkdestroy(tt)
      
    }else{
      if(respuesta == "no"){
        tkconfigure(tt,cursor="watch")
        analysis.window(tclvalue(tkget(txt,"0.0","end")), follow.wizard = FALSE, envir,swap)
        tkdestroy(tt)
      }
    }
  }
  
  topMenu <- tkmenu(tt)
  tkconfigure(tt,menu=topMenu)
  fileMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(fileMenu,"command",label="Quit", command=function(){
    tkdestroy(tt)})
  tkadd(topMenu,"cascade",label="File", menu=fileMenu)
  tkadd(topMenu,"command",label="Back", command = function(){
    tkdestroy(tt)
    genArise()
  })
  
  tkadd(topMenu,"command",label="Analyze", command = function(){
    follow.wizard()
  })
  
  save.as.pdf <-  function(){
    name <- tclvalue(tkgetSaveFile(initialdir = get("path.graphics", envir = envir),
                                   initialfile=get("spot.name",envir=envir),filetypes="{{PDF Files} {.pdf}} {{All files} *}"))
    name <- unlist(strsplit(name, "\\."))[1]
    if(swap == 0){
      if (!nchar(name))
        tkmessageBox(parent = tt,  message= "You must write a name of file!", icon = "error", default = "ok")
      else{
        rbVal <- as.character(tclvalue(rbValue))
        if(rbVal == "Red & Green"){
          pdf(paste(name, "R&G.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "R&G.pdf", sep = "_"))
          imageLimma(log(datos$Cy5, 2) - log(datos$Cy3, 2), row = get("nr", envir = envir), column = get("nc", envir = envir),
                   meta.row = get("nmr", envir = envir),meta.column = get("nmc", envir = envir))
          dev.off(dev.cur())
        }
        else{
          if(rbVal == "Red"){
            pdf(paste(name, "BgCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "BgCy5.pdf", sep = "_"))
            imageLimma(log(datos$BgCy5, 2), get("nr", envir = envir), get("nc", envir = envir),
                       get("nmr", envir = envir),
                       get("nmc", envir = envir), low = "white", high = "red")
            dev.off(dev.cur())
          }
          else{
            pdf(paste(name, "BgCy3.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "BgCy3.pdf", sep = "_"))
            imageLimma(log(datos$BgCy3, 2),get("nr", envir = envir), get("nc", envir = envir),
                       get("nmr", envir = envir),
                       get("nmc", envir = envir), low = "white", high = "green")
            dev.off(dev.cur())
          }
        }
      }
    }
    else if(swap == 1){
      if (!nchar(name))
        tkmessageBox(parent = tt,  message= "You must write a name of file!", icon = "error", default = "ok")
      else{
        pdf(paste(name,"Swap.pdf",sep="_"),horiz=F, height=8,width=8,title=paste(name, "Swap.pdf", sep = "_"))
        swap.plot()
        dev.off(dev.cur())
      }
    }
  }
  
  optionsMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(optionsMenu,"command",label="Save graphic as PDF",command = function(){
    save.as.pdf()})
  tkadd(optionsMenu,"command",label="Note",command = function(){
    note(envir)
  })
  
  tkadd(topMenu,"cascade",label="Options", menu =optionsMenu)
  helpMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(helpMenu,"command",label="About GenArise",command = function() help())
  tkadd(topMenu,"cascade",label="Help",menu = helpMenu)
  upper.frame <- tkframe(tt)
  swap.plot <- function(){
    cond <- get("cond.index",envir=envir)
    R1 <- get("R1",envir=envir)
    R2 <- get("R2",envir=envir)
    par( bg = "black", col.axis = "white",col.lab = "white", fg = "white")
    plot(R1,R2, xlab = "log2(Cy5/Cy3) File1", ylab = "log2(Cy3/Cy5) File2", pch = ".",col = "white")
    points(R1[!cond],R2[!cond],col="blue",pch=".")
  }
  if(swap==0){
    img <- tkrplot(upper.frame, fun = function() imageLimma(M, get("nr", envir = envir), get("nc", envir = envir),
                                  get("nmr", envir = envir),
                                  get("nmc", envir = envir)), hscale= get("Myhscale", envir = envir),vscale = get("Myvscale",
                                                                                                       envir = envir))    
    frameOverall <- tkframe(upper.frame)
    frameUpper <- tkframe(frameOverall,relief="groove",borderwidth=2)
    tkgrid(tklabel(frameUpper, text = "Spot Features", font = "Helvetica 12"), pady = "7")
    tkgrid(tklabel(frameUpper, text = paste("Name:   ", get("spot.name", envir = envir))), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("Rows:           ", get("nr", envir = envir))), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("Columns:       ", get("nc", envir = envir))), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("MetaRows  :     ", get("nmr", envir = envir))), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("MetaColumns:  ", get("nmc", envir = envir))), sticky = "w", pady = "2")
    tkgrid(frameUpper, padx  = "10", pady = "10")
    
  
    radio.frame <- tkframe(frameOverall,relief="groove",borderwidth=2)
    
# diagnostic plots
    rbValue <- tclVar("Red & Green")
    rb1 <- tkradiobutton(radio.frame, text = "Cy5/Cy3", value = "Red & Green", variable = rbValue,
                         command = function() {tkrreplot(img, fun = function() imageLimma(M,
                                                                get("nr", envir = envir),
                                                                get("nc", envir = envir),
                                                                get("nmr", envir = envir),
                                                                get("nmc", envir = envir)))})
    rb2 <- tkradiobutton(radio.frame, text = "Background Cy5", value = "Red", variable = rbValue,
                         command = function() {tkrreplot(img, fun = function()imageLimma(log(datos$BgCy5,2),
                                                                get("nr", envir = envir),
                                                                get("nc", envir = envir),
                                                                get("nmr", envir = envir),
                                                                get("nmc", envir = envir), low = "white", high = "red"))})
    rb3 <- tkradiobutton(radio.frame, text = "Background Cy3", value = "Green", variable = rbValue,
                         command = function() {tkrreplot(img, fun = function()imageLimma(log(datos$BgCy3, 2),
                                                                get("nr", envir = envir),
                                                                get("nc", envir = envir),
                                                                get("nmr", envir = envir),
                                                                get("nmc", envir = envir), low = "white", high = "green"))})
    tkgrid(rb1, sticky = "w", pady = "2",padx="10")
    tkgrid(rb2, sticky = "w", pady = "2",padx="10")
    tkgrid(rb3, sticky = "w", pady = "2",padx="10")
    tkgrid(frameUpper, padx  = "20", pady = "120")
#Label Diagnostic Plots
    tkgrid(tklabel(upper.frame, text = "Diagnostic Plots", font = "Helvetica 12"))  
    tkgrid(radio.frame, padx  = "20", pady = "10")
    tkgrid(img, frameOverall, sticky = "nw")
    tkgrid(tklabel(upper.frame, text = "This plots allow to look at the variation of R and Background values over the array.", font = "Helvetica 10"))  
    tkgrid(upper.frame, pady = "10")
  }
  else if(swap==1){
    img <- tkrplot(upper.frame, fun = function() swap.plot())
    frameOverall <- tkframe(upper.frame)
    frameUpper <- tkframe(frameOverall,relief="groove",borderwidth=2)
    tkgrid(tklabel(frameUpper, text = "Spot Features", font = "Helvetica 12"), pady = "7")
    tkgrid(tklabel(frameUpper, text = get("spot.name", envir = envir)), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("Rows:           ", get("nr.global", envir = envir))), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("Columns:       ", get("nc.global", envir = envir))), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("MetaRows  :     ", get("nmr.global", envir = envir))), sticky = "w", pady = "2")
    tkgrid(tklabel(frameUpper, text = paste("MetaColumns:  ", get("nmc.global", envir = envir))), sticky = "w", pady = "2")
    tkgrid(frameUpper, padx  = "10", pady = "10")

    radio.frame <- tkframe(frameOverall,relief="groove",borderwidth=2)
    tkgrid(tklabel(radio.frame, text = get("conf", envir = envir)), sticky = "w", pady = "2")
    tkgrid(tklabel(radio.frame, text = get("disp", envir = envir)), sticky = "w", pady = "2")
    tkgrid(radio.frame, padx  = "20", pady = "10")
    tkgrid(img, frameOverall, sticky = "nw")
    tkgrid(upper.frame, pady = "10")
  }
  area.frame <- tkframe(tt,relief="groove")
  yscr <- tkscrollbar(area.frame, repeatinterval = 5,
                      command=function(...)tkyview(txt,...))
  txt <- tktext(area.frame,bg="white",font="courier",width =get("barWidth",envir=envir),height=get("barHeight",envir=envir),yscrollcommand=function(...)tkset(yscr,...))
  tkgrid(txt, yscr, sticky = "w")
  tkgrid.configure(yscr,rowspan=4,sticky="ns")
  tkinsert(txt,"end","GenArise Microarray Analyzer\n")
  tkinsert(txt,"end","Institute of Cellular Physiology UNAM")
  tkconfigure(txt, state="disabled")
  tkgrid(area.frame, padx = "10", pady = "10", sticky = "w")
}

# Auxiliar function to call the Zscore.plot function in the GUI
Zscore.points <-  function(type="ri",text, envir,swap){
  tt <- tktoplevel()
  tkfocus(tt)
  tkwm.title(tt,"GenArise Microarray Analyzer")
  topMenu <- tkmenu(tt)
  tkconfigure(tt,menu=topMenu)
  fileMenu <- tkmenu(topMenu,tearoff=FALSE)
  upper.frame <- tkframe(tt)
  sd.frame <- tkframe(upper.frame, borderwidth=2)
  ratio.buttons.frame <- tkframe(sd.frame,relief="groove",borderwidth=2)
  dist  <- tclVar(5)
  
  radio1 <- tkradiobutton(ratio.buttons.frame, text="Zscore < 1",
                          value=1, variable=dist, command=function(){
                            tkrreplot(img, fun = function()
                                      Zscore.plot(get("Zscore.spot", envir = envir), all = FALSE, Zscore.max = 1, col = "green"))})
  radio2 <- tkradiobutton(ratio.buttons.frame, text="1 < Zscore < 1.5",
                          value=2, variable=dist,command = function(){
                            tkrreplot(img, fun = function() Zscore.plot(get("Zscore.spot", envir = envir),
                                             Zscore.max = 1.5, Zscore.min = 1, all = FALSE, col = "blue"))})
  radio3 <- tkradiobutton(ratio.buttons.frame, text="1.5 < Zscore < 2",
                          value=3, variable=dist,command = function(){
                            tkrreplot(img, fun = function() Zscore.plot(get("Zscore.spot", envir = envir),
                                             Zscore.max = 2, Zscore.min = 1.5, all = FALSE, col = "cyan"))})
  radio4 <- tkradiobutton(ratio.buttons.frame, text="Zscore > 1.5",
                          value=4, variable=dist,command = function(){
                            tkrreplot(img, fun = function() Zscore.plot(get("Zscore.spot", envir = envir),
                                             Zscore.min = 1.5, all = FALSE, col = "yellow"))})
  radio5 <- tkradiobutton(ratio.buttons.frame, text="Zscore > 2",
                          value=5, variable=dist, command=function(){
                            tkrreplot(img, fun = function() Zscore.plot(get("Zscore.spot", envir = envir),
                                             Zscore.min = 2, all = FALSE, col = "snow"))})
  radio6 <- tkradiobutton(ratio.buttons.frame, text="All",
                          value=6, variable=dist, command=function(){
                            tkrreplot(img, fun = function() Zscore.plot(get("Zscore.spot", envir = envir)))})
  
  tkgrid(radio1, sticky="w")
  tkgrid(radio2, sticky="w")
  tkgrid(radio3, sticky="w")
  tkgrid(radio4, sticky="w")
  tkgrid(radio5, sticky="w")
  tkgrid(radio6, sticky="w")

  # get the information for the annotation function
  annotations.but <- tkbutton(sd.frame,text = "Annotations", command=function(){
    choose.specie <- tktoplevel()
    tkfocus(choose.specie)
    tkwm.title(choose.specie,"Specie Selector")
    tkgrid(tklabel(choose.specie,text="Choose the specie"))
    species <- c("Mus musculus","Saccharomyces Cerevisiae","Rattus norvegicus", "NCBI")
    comboBox <- tkwidget(choose.specie, "ComboBox",editable=FALSE,values=species)
    tkgrid(comboBox)
    
    OnOK <- function()
      {
        datos <- attr(get("Zscore.spot", envir = envir), "dataSets")
        val <- as.numeric(tclvalue(dist))
        if(val != 6){
          Zscore <- datos$Zscore
          if(val == 1){
            
            indexUp <- Zscore > 1
            indexDown <- Zscore < -1
            name <- "_less_1SD"
          }
          if(val == 2){
            indexUp <- Zscore > 1 & Zscore < 1.5
            indexDown <- Zscore < -1 & Zscore > -1.5
            name <- "_1-1.51SD"
          }
          if(val == 3){
            indexUp <- Zscore > 1.5 & Zscore < 2
            indexDown <- Zscore < -1.5 & Zscore > -2
            name <- "_1_5-2SD"
          }
          if(val == 4){
            indexUp <- Zscore > 1.5
            indexDown <- Zscore < -1.5
            name <- "_greater_1_5SD"
          }
          if(val == 5){
          indexUp <- Zscore > 2
          indexDown <- Zscore < -2
          name <- "_greater_2SD"
        }
          if(!is.null(datos$Symdesc)){
            Up <- list(Cy3 = datos$Cy3[indexUp], Cy5 = datos$Cy5[indexUp],
                       Id = datos$Id[indexUp], Symdesc = datos$Symdesc[indexUp], Zscore = datos$Zscore[indexUp])
            Down <- list(Cy3 = datos$Cy3[indexDown], Cy5 = datos$Cy5[indexDown],
                         Id = datos$Id[indexDown], Symdesc = datos$Symdesc[indexDown], Zscore = datos$Zscore[indexDown])
          }
          else{
            Up <- list(Cy3 = datos$Cy3[indexUp], Cy5 = datos$Cy5[indexUp],
                       Id = datos$Id[indexUp], Zscore = datos$Zscore[indexUp])
            Down <- list(Cy3 = datos$Cy3[indexDown], Cy5 = datos$Cy5[indexDown],
                         Id = datos$Id[indexDown], Zscore = datos$Zscore[indexDown])
          }
          kk <- as.data.frame(Up)
          kk2 <- as.data.frame(Down)
          kk <- apply(kk, 2, as.vector)
          kk2 <- apply(kk2, 2, as.vector)
          la.especie <- as.numeric(tclvalue(tcl(comboBox,"getvalue")))
          if(!is.null(datos$Symdesc)){
            annotations(kk, species[(la.especie + 1)], 3, symbol=4, file.path(get("path.project", envir = envir), paste("annotationsUp",name,".htm",sep="")))
            annotations(kk2, species[(la.especie + 1)], 3, symbol=4, file.path(get("path.project", envir = envir), paste("annotationsDown",name,".htm",sep="")))
          }
          else{
            annotations(kk, species[(la.especie + 1)], 3, symbol=NULL, file.path(get("path.project", envir = envir), paste("annotationsUp",name,".htm",sep="")))
            annotations(kk2, species[(la.especie + 1)], 3, symbol=NULL,file.path(get("path.project", envir = envir), paste("annotationsDown",name,".htm",sep="")))
          }
          tkdestroy(choose.specie)
         }
        else{
          kk <- as.data.frame(datos)
          kk <- apply(kk, 2, as.vector)
          la.especie <- as.numeric(tclvalue(tcl(comboBox,"getvalue")))
          if(!is.null(datos$Symdesc))
            annotations(kk, species[(la.especie + 1)], 3, 4, file.path(get("path.project", envir = envir), "annotationsAll.htm"))
          else
            annotations(kk, species[(la.especie + 1)], 3,symbol=NULL, file.path(get("path.project", envir = envir), "annotationsAll.htm"))
          tkdestroy(choose.specie)
         }
      }
    
    OnCancel <-  function(){
      tkdestroy(choose.specie)
    }
    
    OK.but <-tkbutton(choose.specie,text="   OK   ",command=OnOK)
    Cancel.but <-tkbutton(choose.specie,text="   Cancel   ",command=OnCancel)
    tkgrid(OK.but, Cancel.but, padx = "20", pady = "10")
    tkfocus(tt)
  })
  
  tkgrid(ratio.buttons.frame, sticky="w", pady = "20")
  tkgrid(annotations.but, sticky="w", pady = "10")
  
  tkselect(radio6)
  tkadd(fileMenu,"command",label="Close",command=function(){
    tkdestroy(tt)
    genArise.init(envir)})
  tkadd(fileMenu,"command",label="Exit",command=function(){
    tkdestroy(tt)})
  tkadd(topMenu,"cascade",label="File",menu=fileMenu)
  tkadd(topMenu,"command",label="Back", command = function(){
    sure <- back.gui(envir)
    if(sure == 0){
      tkdestroy(tt)
      if(swap== 0){
        principal(envir,0)
      }
      else if(swap == 1){
        principal(envir,1)
      }
  }})
  
  optionsMenu <- tkmenu(topMenu,tearoff=FALSE)
  
  make.file.name <- function(file.type, ext){
    switch(as.integer(tclvalue(dist)),
           suggested.name <- paste(get("spot.name",envir=envir), "_lower_1.", ext, sep = ""),
           suggested.name <- paste(get("spot.name",envir=envir), "_1-1_5.", ext, sep = ""),
           suggested.name <- paste(get("spot.name",envir=envir), "_1_5-2.", ext, sep = ""),
           suggested.name <- paste(get("spot.name",envir=envir), "_greater_1_5.", ext, sep = ""),
           suggested.name <- paste(get("spot.name",envir=envir), "_greater_2.", ext, sep = ""),
           suggested.name <- paste(get("spot.name",envir=envir), "_all.", ext, sep = ""),
           )
    if(ext == "pdf"){
      salida <- "path.graphics"
    }else{
      salida <-  "path.results"
    }
    tclvalue(tkgetSaveFile(initialdir = get(salida, envir = envir),
                           initialfile = suggested.name, filetypes = file.type))


    
  }    
  tkadd(optionsMenu,"command",label="Save graphic as PDF",command = function() {
    name <- make.file.name(file.type = "{{PDF Files} {.pdf}} {{All files} *}", ext ="pdf")
    name <- unlist(strsplit(name, "\\."))[1]
    if (!nchar(name))
      tkmessageBox(parent = tt,  message= "You must write a name of file!", icon = "error", default = "ok")
    else{
      pdf(paste(name,".pdf",sep=""), horiz = FALSE, height = 8, width = 8, title = name)
      switch(as.integer(tclvalue(dist)),
             Zscore.plot(get("Zscore.spot", envir = envir), all = FALSE, Zscore.max = 1, col = "green"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.max = 1.5, Zscore.min = 1, all = FALSE, col = "blue"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.max = 2, Zscore.min = 1.5, all = FALSE, col = "cyan"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.min = 1.5, all = FALSE, col = "yellow"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.min = 2, all = FALSE, col = "snow"),
             Zscore.plot(get("Zscore.spot", envir = envir)))
      dev.off(dev.cur())
    }
  })

  # save the Zscore for the data set in a file
  tkadd(optionsMenu,"command",label="Write as outputfile",command = function() {
    name <- make.file.name(file.type = "{{TXT Files} {.txt}} {{All files} *}", ext="txt")
    if (!nchar(name))
      tkmessageBox(parent = tt,  message= "You must write a name of file!", icon = "error", default = "ok")
    else{
      val <- as.integer(tclvalue(dist))
      if(swap==0){
        if( val == 6){
          if(is.null(attr(get("Zscore.spot", envir = envir),"dataSets")$Symdesc))
            write.table("Cy3\tCy5\tId\tZscore",name, quote = FALSE,sep = "\t", col.names = FALSE, row.names = FALSE )
          else
            write.table("Cy3\tCy5\tId\tSymbol\tZscore",name, quote = FALSE,sep = "\t", col.names = FALSE, row.names = FALSE )
          write.table(attr(get("Zscore.spot", envir = envir), "dataSets"),
                      name, quote = FALSE, col.names = FALSE, row.names = FALSE,sep="\t",append=TRUE)
        }else if(val == 5){
          write.dataSet(get("Zscore.spot", envir = envir), name, Zscore.min = 2)
        }
        else if(val == 4){
          write.dataSet(get("Zscore.spot", envir = envir), name, Zscore.min = 1.5)
        }
        else if(val == 3){
          write.dataSet(get("Zscore.spot", envir = envir), name, Zscore.min = 1.5, Zscore.max = 2)
        }
        else if(val == 2){
          write.dataSet(get("Zscore.spot", envir = envir), name, Zscore.min = 1, Zscore.max = 1.5)
        }
        else if(val == 1){
          write.dataSet(get("Zscore.spot", envir = envir), name, Zscore.max = 1)
        }
      }
      else{
        warn <- tktoplevel()
        tkfocus(warn)
        message <- tklabel(warn,text="This operation may take a few minutes")
        tkwm.title(warn,"Note")
        tkgrid(message)
        tkfocus(warn)
        Sys.sleep(1)
        if(get("ind.norm", envir = envir) == 0){
          single.norm(envir)
          assign("ind.norm",1,envir=envir)
        }
        Zscore.spot <- get("Zscore.spot", envir = envir)
        if( val == 6){
            get.Zscore(Zscore.spot, name, Zscore.min = 0, all=TRUE, envir = envir)
        }else if(val == 5){
            get.Zscore(Zscore.spot, name, Zscore.min = 2, envir = envir)
        }
        else if(val == 4){
            get.Zscore(Zscore.spot, name, Zscore.min = 1.5, envir = envir)
        }
        else if(val == 3){
            get.Zscore(Zscore.spot, name, Zscore.min = 1.5, Zscore.max = 2, envir = envir)
        }
        else if(val == 2){
            get.Zscore(Zscore.spot, name, Zscore.min = 1, Zscore.max = 1.5, envir = envir)
        }
        else if(val == 1){
            write.dataSet(Zscore.spot, name, Zscore.max = 1)
        }
        tkdestroy(warn)
      }
    }
  })
  
  tkadd(optionsMenu,"command",label="Notes",command = function()note(envir))
  tkadd(topMenu,"cascade",label="Options",menu =optionsMenu)
  
  helpMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(helpMenu,"command",label="About genArise...", command=function() help())
  tkadd(topMenu,"cascade",label="Help",menu = helpMenu)
  
  
  frame.label <- tkframe(tt,relief="groove",borderwidth=2)

  #######################

  lower.menu <- tkframe(tt, relief = "groove", borderwidth=2)
  
  image.menu.frame <- tkframe(lower.menu)
  
  tcl("image","create","photo","acroread",file= file.path(get("icons.dir", envir = envir),"icons/acroread.png"))
  acroread.button <- tkbutton(image.menu.frame,image="acroread", command = function(){
    name <- make.file.name(file.type = "{{PDF Files} {.pdf}} {{All files} *}", ext ="pdf")
    name <- unlist(strsplit(name, "\\."))[1]
    if (!nchar(name))
      tkmessageBox(parent = tt,  message= "You must write a name of file!", icon = "error", default = "ok")
    else{
      pdf(paste(name,".pdf",sep=""), horiz = FALSE, height = 8, width = 8, title = name)
      switch(as.integer(tclvalue(dist)),
             Zscore.plot(get("Zscore.spot", envir = envir), all = FALSE, Zscore.max = 1, col = "green"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.max = 1.5, Zscore.min = 1, all = FALSE, col = "blue"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.max = 2, Zscore.min = 1.5, all = FALSE, col = "cyan"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.min = 1.5, all = FALSE, col = "yellow"),
             Zscore.plot(get("Zscore.spot", envir = envir), Zscore.min = 2, all = FALSE, col = "snow"),
             Zscore.plot(get("Zscore.spot", envir = envir)))
      dev.off(dev.cur())
    }
  }) 
  tkconfigure(acroread.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","editor",file=file.path(get("icons.dir", envir = envir),"icons/editor.png"))
  editor.button <- tkbutton(image.menu.frame,image="editor", command = function() note(envir))
  tkconfigure(editor.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","back",file=file.path(get("icons.dir", envir = envir),"icons/regresar.png"))
  back.button <- tkbutton(image.menu.frame,image="back",command =  function(){
     sure <- back.gui(envir)
    if(sure == 0){
      tkdestroy(tt)
      if(swap== 0){
        principal(envir,0)
      }
      else if(swap == 1){
        principal(envir,1)
      }
  }})
  tkconfigure(back.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","help",file=file.path(get("icons.dir", envir = envir),"icons/help.png"))
  help.button <- tkbutton(image.menu.frame,image="help", command = function() help())
  tkconfigure(help.button, heigh = 16, width = 16)

  tcl("image","create","photo","exit",file=file.path(get("icons.dir", envir = envir),"icons/logout.png"))
  exit.button <- tkbutton(image.menu.frame,image="exit", command = function(){
    tkdestroy(tt)})
  tkconfigure(exit.button, heigh = 16, width = 16)
  
  tkgrid(acroread.button, editor.button, back.button, help.button, exit.button, sticky = "nw")
  
  etiqueta <- tklabel(lower.menu, text = "")
  tkconfigure(etiqueta, text = "", width = "90") 
  tkgrid(image.menu.frame, etiqueta, sticky = "w")
  tkgrid(lower.menu)


  #######################
  tkfocus(tt)
  img <- tkrplot(upper.frame,fun = function()Zscore.plot(get("Zscore.spot", envir = envir),type),
                 hscale= get("Myhscale", envir = envir),vscale = get("Myvscale",envir = envir))
  tkgrid(img,sd.frame, padx = "10", pady = "10")
  tkgrid(upper.frame, pady = "10")
  area.frame <- tkframe(tt,relief="groove")
  yscr <- tkscrollbar(area.frame, repeatinterval = 5,
                      command=function(...)tkyview(txt,...))
  
 
  txt <- tktext(area.frame,bg="white",font="courier",width =get("barWidth",envir=envir),height=get("barHeight",envir=envir),yscrollcommand=function(...)tkset(yscr,...))
  tkgrid(txt, yscr)
  tkgrid.configure(yscr,rowspan=4,sticky="nsw")
  tkinsert(txt,"end",text)
  tkinsert(txt,"end","\n\nZscore Done!!")
  tkconfigure(txt, state="disabled")
  tkgrid(area.frame, padx = "10", sticky = "w")
}

# Window for the analysis
analysis.window <-  function(texto, follow.wizard = FALSE, envir,swap=0){
if(swap==1){
  assign("ind.norm",0, envir=envir)
}else if(swap==0){
  assign("corrected", 0, envir = envir)
}
  # check if the background correction will be done
  bg.question <- function(){
    answer <-tkmessageBox(message="Do you want to make background correction?",icon="question",type="yesnocancel",default="yes")
    ans <- as.character(answer)
    op.counter <<- op.counter + 1
    write.spot(get("o.spot", envir = envir), paste(get("path.results", envir = envir),
                               .Platform$file.sep, "original.txt",sep = ""))
    set.history.project(get("history.project", envir = envir), "Original Spot", "original.txt")
    
    if(ans == "yes"){
      c.spot <- bg.correct(get("a.spot", envir = envir))
      op.counter <<- op.counter + 1
      tkconfigure(txt, state="normal")
      tkinsert(txt,"end","\n\nBackground correction done.............")
      tkconfigure(txt, state="disable")
      assign("c.spot", c.spot, envir = envir)
      write.spot(get("c.spot", envir = envir), paste(get("path.results", envir = envir),
                                 .Platform$file.sep, "corrected.txt",sep = ""))
#      print(attr(get("c.spot", envir = envir),"spotData")$Symdesc)
      set.history.project(get("history.project", envir = envir), "Corrected Spot", "corrected.txt")
      assign("a.spot", c.spot, envir = envir)
      tkpack(corr.radio,anchor="w",padx="10")
      tkselect(corr.radio)
      assign("corrected",1, envir = envir)
    }
    else {
      if(ans == "no"){
        assign("corrected",1, envir = envir)
      }
      else{
        assign("corrected",2, envir = envir)      }
    }
  }

  # GUI behavior after the data normalization
  normalized.gui <- function(type = 1, history.entry, analysis.plot){ # 1 by grid, 2 global
    if(swap==0){
    if(get("corrected",envir=envir) == 0) bg.question()
    if(get("corrected",envir=envir) == 2) assign("corrected",0,envir=envir)
    else{
      if(get("corrected",envir = envir) == 1){
        tkconfigure(tt,cursor="watch")
        switch(type,
               {n.spot <- grid.norm(get("a.spot", envir = envir),
                                    get("nr", envir = envir), get("nc", envir = envir))
                op.counter <<- op.counter + 1
                history.text <- "\n\nNormalization by grid done!..."},
               {n.spot <- global.norm(get("a.spot", envir = envir))
                history.text <- "\n\nGlobal normalization done!..."})
        tkconfigure(history.entry, state="normal")
        tkinsert(history.entry,"end",history.text)
        tkconfigure(history.entry, state="disable")
        assign("n.spot", n.spot, envir = envir)
        write.spot(get("n.spot", envir = envir), paste(get("path.results", envir = envir),
                                   .Platform$file.sep, "normalized.txt",sep = ""))
        set.history.project(get("history.project", envir = envir), "Normalized Spot", "normalized.txt")
        assign("a.spot", n.spot, envir = envir)
        tkrreplot(analysis.plot,fun=function()graphic.choose(get("a.spot", envir = envir),
                                  get("graphic.type", envir = envir)))
        tkpack(normal.radio,anchor="w",padx="10")
        tkselect(normal.radio)
        tkentryconfigure(topMenu, "4", state ="disable" )
        tkconfigure(normal.button, state = "disable")
        tkconfigure(tt,cursor="arrow")
      }
    }
  }
    else if(swap==1){
 tkconfigure(tt,cursor="watch")
        switch(type,
               {n.spot <- grid.norm(get("a.spot", envir = envir),
                                    get("nr.global", envir = envir), get("nc.global", envir = envir))
                op <- get("op.counter", envir = envir) + 1
                assign("op.counter", op, envir = envir)
                history.text <- "\n\nNormalization by grid done!..."},
               {n.spot <- global.norm(get("a.spot", envir = envir))
                history.text <- "\n\nGlobal normalization done!..."})
        tkconfigure(history.entry, state="normal")
        tkinsert(history.entry,"end",history.text)
        tkconfigure(history.entry, state="disable")
        assign("n.spot", n.spot, envir = envir)
        write.spot(get("n.spot", envir = envir), paste(get("path.results", envir = envir),
                                   .Platform$file.sep, "normalized.txt",sep = ""))
        set.history.project(get("history.project", envir = envir), "Normalized Spot", "normalized.txt")
        assign("a.spot", n.spot, envir = envir)
        tkrreplot(analysis.plot,fun=function()graphic.choose(get("a.spot", envir = envir),
                                  get("graphic.type", envir = envir)))
        tkpack(normal.radio,anchor="w",padx="10")
        tkselect(normal.radio)
        tkentryconfigure(topMenu, "4", state ="disable" )
        tkconfigure(normal.button, state = "disable")
        tkconfigure(tt,cursor="arrow")
}
  }

  # GUI behavior after the data filtering
  filter.gui <-  function(history.text, analysis.plot){
    if(swap==0){
    if(get("corrected",envir=envir) == 0) bg.question()
    if(get("corrected",envir=envir) == 2) assign("corrected",0,envir=envir)
       else{
         tkconfigure(tt,cursor="watch")
         f.spot <- filter.spot(get("a.spot", envir = envir))
         op.counter <<- op.counter + 1
         tkconfigure(history.text, state="normal")
                                        #    tkinsert(history.text,"end","\n\nIn some cases, the correlation of a signal between log background and log signal is observed to be larger than that of the other signal in the experiment. If this is the case, you could see a strange effect after filter by intensity.\n\nIntensity-based filtering done.............")
         tkinsert(history.text,"end","\n\nIn some cases, the standard deviation of the background of a signal is observed to be much larger than that of the other signal in the experiment. If this is the case, you could see a strange effect in the plot after filter by intensity.\n\nIntensity-based filtering done.............")
         tkconfigure(history.text, state="disable")
         assign("f.spot", f.spot, envir = envir)
         write.spot(get("f.spot", envir = envir), paste(get("path.results", envir = envir),
                                    .Platform$file.sep, "filter.txt",sep = ""))
         set.history.project(get("history.project", envir = envir), "Filtered Spot", "filter.txt")
         assign("a.spot", f.spot, envir = envir)
         tkrreplot(analysis.plot,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))
         tkpack(fil.radio,anchor="w",padx="10")
         tkselect(fil.radio)
         tkentryconfigure(filterMenu, "0", state ="disable" )
         tkconfigure(filter.button, state = "disable")
         count.filter <<- count.filter +1
         tkentryconfigure(normalMenu, "0", state ="disable" )
         if(count.filter == 2){
           tkentryconfigure(topMenu, "3", state ="disable" )}
         tkconfigure(tt,cursor="arrow")
       }
  }
    else if(swap==1){
 tkconfigure(tt,cursor="watch")
         f.spot <- filter.spot(get("a.spot", envir = envir))
         op <- get("op.counter", envir = envir) + 1
         assign("op.counter", op, envir = envir)
         tkconfigure(history.text, state="normal")
                                        #    tkinsert(history.text,"end","\n\nIn some cases, the correlation of a signal between log background and log signal is observed to be larger than that of the other signal in the experiment. If this is the case, you could see a strange effect after filter by intensity.\n\nIntensity-based filtering done.............")
         tkinsert(history.text,"end","\n\nIn some cases, the standard deviation of the background of a signal is observed to be much larger than that of the other signal in the experiment. If this is the case, you could see a strange effect in the plot after filter by intensity.\n\nIntensity-based filtering done.............")
         tkconfigure(history.text, state="disable")
         assign("f.spot", f.spot, envir = envir)
         write.spot(get("f.spot", envir = envir), paste(get("path.results", envir = envir),
                                    .Platform$file.sep, "filter.txt",sep = ""))
         set.history.project(get("history.project", envir = envir), "Filtered Spot", "filter.txt")
         assign("a.spot", f.spot, envir = envir)
         tkrreplot(analysis.plot,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))
         tkpack(fil.radio,anchor="w",padx="10")
         tkselect(fil.radio)
         tkentryconfigure(filterMenu, "0", state ="disable" )
         tkconfigure(filter.button, state = "disable")
         count.filter <<- count.filter +1
         tkentryconfigure(normalMenu, "0", state ="disable" )
         if(count.filter == 2){
           tkentryconfigure(topMenu, "3", state ="disable" )}
         tkconfigure(tt,cursor="arrow")
}
  }
     

  
  # GUI behavior after remove replicates
  # 1 mean replicate filtering, 2 non-extreme values replicate, 3 geometric mean
  remove.duplicates.gui <-  function(type = 1, history.entry, analysis.plot){
    if(swap==0){
    if(get("corrected",envir=envir) == 0) bg.question()
    if(get("corrected",envir=envir) == 2) assign("corrected",0,envir=envir)
    else{
      if(get("corrected",envir=envir) == 1){
        tkconfigure(tt,cursor="watch")
        switch(type,
               {u.spot <- meanUnique(get("a.spot", envir = envir))
                op.counter <<- op.counter + 1
                history.text <-  "\n\nMean replicates filtering done!..."
                history.project.text <- "Mean replicates filter"},
               {u.spot <- alter.unique(get("a.spot", envir = envir))
                op.counter <<- op.counter + 1
                history.text <- "\n\nNon-extreme values replicate"
                history.project.text <- "Non-extreme filter dup"},
               {u.spot <- spotUnique(get("a.spot", envir = envir))
                op.counter <<- op.counter + 1
                history.text <- "\n\nGeometric mean filtering done!..."
                history.project.text <- "Geometric mean filter dup"})
        tkconfigure(history.entry, state="normal")
        tkinsert(history.entry,"end", history.text)
        tkconfigure(history.entry, state="disable")
        assign("u.spot", u.spot, envir = envir)
        write.spot(get("u.spot", envir = envir), paste(get("path.results", envir = envir),
                                   .Platform$file.sep, "withoutDup.txt",sep = ""))
        set.history.project(get("history.project", envir = envir), history.project.text, "withoutDup.txt")
        assign("a.spot", u.spot, envir = envir)
        tkrreplot(analysis.plot,fun=function()graphic.choose(get("a.spot", envir = envir),
                                  get("graphic.type", envir = envir)))
        tkpack(uniq.radio,anchor="w",padx="10")
        tkselect(uniq.radio)
        tkentryconfigure(filterMenu, "1", state ="disable" )
        count.filter <- count.filter + 1
        tkentryconfigure(normalMenu, "0", state ="disable" )
        if(count.filter == 2){
          tkentryconfigure(topMenu, "3", state ="disable" )}}
      tkconfigure(tt,cursor="arrow")
    }
  }
    else if(swap==1){
        tkconfigure(tt,cursor="watch")
        switch(type,
               {u.spot <- meanUnique(get("a.spot", envir = envir))
                op <- get("op.counter", envir = envir) + 1
                assign("op.counter", op, envir = envir)
                history.text <-  "\n\nMean replicates filtering done!..."
                history.project.text <- "Mean replicates filter"},
               {u.spot <- alter.unique(get("a.spot", envir = envir))
                op <- get("op.counter", envir = envir) + 1
                assign("op.counter", op, envir = envir)
                history.text <- "\n\nNon-extreme values replicate"
                history.project.text <- "Non-extreme filter dup"},
               {u.spot <- spotUnique(get("a.spot", envir = envir))
                op <- get("op.counter", envir = envir) + 1
                assign("op.counter", op, envir = envir)
                history.text <- "\n\nGeometric mean filtering done!..."
                history.project.text <- "Geometric mean filter dup"})
        tkconfigure(history.entry, state="normal")
        tkinsert(history.entry,"end", history.text)
        tkconfigure(history.entry, state="disable")
        assign("u.spot", u.spot, envir = envir)
        write.spot(get("u.spot", envir = envir), paste(get("path.results", envir = envir),
                                   .Platform$file.sep, "withoutDup.txt",sep = ""))
        set.history.project(get("history.project", envir = envir), history.project.text, "withoutDup.txt")
        assign("a.spot", u.spot, envir = envir)
        tkrreplot(analysis.plot,fun=function()graphic.choose(get("a.spot", envir = envir),
                                  get("graphic.type", envir = envir)))
        tkpack(uniq.radio,anchor="w",padx="10")
        tkselect(uniq.radio)
        tkentryconfigure(filterMenu, "1", state ="disable" )
        count.filter <- count.filter + 1
        tkentryconfigure(normalMenu, "0", state ="disable" )
        if(count.filter == 2){
          tkentryconfigure(topMenu, "3", state ="disable" )}
      tkconfigure(tt,cursor="arrow")
      }
  }

  # GUI behavior after perform Zscore
  zscore.gui <- function(){
    tkconfigure(tt,cursor="watch")
    assign("Zscore.spot", Zscore(get("a.spot",envir=envir)), envir = envir)
    write.zscore(get("Zscore.spot", envir = envir), file.path(get("path.results", envir = envir), "zscore.txt"))
    set.history.project(get("history.project", envir = envir), "RI Zscore", "zscore.txt")
    Zscore.points(type="ri",text=tclvalue(tkget(txt,"0.0","end")), envir,swap)
    tkdestroy(tt)
  }
  
  count.filter <-  0
  tt <- tktoplevel()
tkfocus(tt)
  tkwm.title(tt,"GenArise Microarray Analyzer")
  topMenu <- tkmenu(tt)
  tkconfigure(tt,menu=topMenu)
  fileMenu <- tkmenu(topMenu,tearoff=FALSE)
  upper.frame <- tkframe(tt)
  frame1 <- tkframe(upper.frame,relief="groove",borderwidth=2)
  dist  <- tclVar(1)
  ori.radio <- tkradiobutton(frame1, text="Original Spot",
                             value=1, variable=dist, command=function(){
                               assign("a.spot", get("o.spot", envir = envir), envir = envir)
                               tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))})
  corr.radio <- tkradiobutton(frame1, text="Corrected Spot",
                              value=2, variable=dist,command = function(){
                                assign("a.spot", get("c.spot", envir = envir), envir = envir)
                                tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))})
  
  normal.radio <- tkradiobutton(frame1, text="Normal Spot",
                                value=3, variable=dist,command = function(){
                                  assign("a.spot", get("n.spot", envir = envir), envir = envir)
                                  tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))})
  
  fil.radio <- tkradiobutton(frame1, text="Filter Spot",
                             value=4, variable=dist,command = function(){
                               assign("a.spot", get("f.spot", envir = envir), envir = envir)
                               tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))})
  uniq.radio <- tkradiobutton(frame1, text="Without duplicates",
                              value=5, variable=dist,command = function(){
                                assign("a.spot", get("u.spot", envir = envir), envir = envir)
                                tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))})
  tkpack(ori.radio,anchor="w",padx="10")
  
  tkadd(fileMenu,"command",label="Close",command=function(){
    tkdestroy(tt)
    genArise.init(envir) })
  tkadd(fileMenu,"command",label="Exit",command=function(){
    tkdestroy(tt)})
  tkadd(topMenu,"cascade",label="File",menu=fileMenu)

  tkadd(topMenu,"command",label="Back", command = function(){
    sure <- back.gui(envir)
    if(sure == 0){
      tkdestroy(tt)
      if(swap== 0){
        principal(envir,0)
      }
      else if(swap == 1){
        principal(envir,1)
      }
  }})
  filterMenu <- tkmenu(topMenu,tearoff=FALSE)
  
  tkadd(filterMenu,"command",label="By Intensity", state ="active",command = function() {
    filter.gui(txt, img)})
#  tkentryconfigure(filterMenu, "0", state ="disable" ) #Inactive temporal
  duplicatesMenu <- tkmenu(filterMenu,tearoff=FALSE)
if(swap==0){
  tkadd(duplicatesMenu,"command",label="Mean Replicate Filtering", command = function(){
    remove.duplicates.gui(1,txt,img)})
  tkadd(duplicatesMenu,"command",label="Non-extreme values replicate filtering", command = function(){
    remove.duplicates.gui(2, txt, img)})
  tkadd(duplicatesMenu,"command",label="Geometric Mean Filter", command = function(){
    remove.duplicates.gui(3, txt, img)})

  tkadd(filterMenu,"cascade",label="Duplicates Analysis", state ="active",menu= duplicatesMenu)
  
  tkadd(topMenu,"cascade",label="Filter",menu =filterMenu)
  
  normalMenu <- tkmenu(topMenu,tearoff=FALSE)
  
  tkadd(normalMenu,"command",label="By grid", state ="active", command=function(){
    normalized.gui(1,txt, img)
  })
  tkadd(normalMenu,"command",label="Global", state ="active", command=function(){
    normalized.gui(2,txt, img)
  })
}
else if(swap==1){
   tkadd(duplicatesMenu,"command",label="Mean Replicate Filtering", command = function(){
    remove.duplicates.gui(1,txt,img); assign("unique.alg", 1, envir = envir)})
  tkadd(duplicatesMenu,"command",label="Non-extreme values replicate filtering", command = function(){
    remove.duplicates.gui(2, txt, img); assign("unique.alg", 2, envir = envir)})
  tkadd(duplicatesMenu,"command",label="Geometric Mean Filter", command = function(){
    remove.duplicates.gui(3, txt, img); assign("unique.alg",3, envir = envir)})

    tkadd(filterMenu,"cascade",label="Duplicates Analysis", state ="active",menu= duplicatesMenu)
  
  tkadd(topMenu,"cascade",label="Filter",menu =filterMenu)
  
  normalMenu <- tkmenu(topMenu,tearoff=FALSE)
  
  tkadd(normalMenu,"command",label="By grid", state ="active", command=function(){
    normalized.gui(1,txt, img)
    assign("norm.alg", 1, envir = envir)
  })
  tkadd(normalMenu,"command",label="Global", state ="active", command=function(){
    normalized.gui(2,txt, img)
    assign("norm.alg", 2, envir = envir)
  })
 }
  

  cys.plot <-  function(){
    assign("graphic.type", 1, envir = envir)
    tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))}

  tkadd(topMenu,"cascade",label="Normalize",menu = normalMenu)
  graphicMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(graphicMenu,"command",label="log(Cy3) vs log(Cy5)",command = function() {
    cys.plot()})
  tkadd(graphicMenu,"command",label="R vs I",command = function() {
    assign("graphic.type", 2, envir = envir)
    tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))})
  tkadd(graphicMenu,"command",label="M vs A",command = function() {
    assign("graphic.type", 3, envir = envir)
    tkrreplot(img,fun=function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)))})
  tkadd(topMenu,"cascade",label="Graphics",menu =graphicMenu)
  
  ZscoreMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(topMenu,"cascade",label="Zscore",menu = ZscoreMenu)
  tkadd(ZscoreMenu,"command",label="R vs I", state="active", command=function(){
    zscore.gui()
  })
  tkadd(ZscoreMenu,"command",label="M vs A", state="active",command=function(){
    assign("Zscore.spot", Zscore(get("a.spot",envir=envir), type = "ma"), envir = envir)
    write.zscore(get("Zscore.spot", envir = envir),file.path(get("path.results", envir = envir), "zscore.txt"))
    set.history.project(get("history.project", envir = envir), "MA Zscore", "zscore.txt")
    Zscore.points(type="ma",text=tclvalue(tkget(txt,"0.0","end")), envir,swap)
    tkdestroy(tt)
  })
  
  optionsMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(optionsMenu,"command",label="Save graphic as PDF",command = function(){
    name <- tclvalue(tkgetSaveFile(initialdir = get("path.graphics", envir = envir),
                                   initialfile=get("spot.name",envir=envir),
                                   filetypes="{{PDF Files} {.pdf}} {{All files} *}"))
    name <- unlist(strsplit(name, "\\."))[1]
    if (!nchar(name))
      tkmessageBox(parent = tt,  message= "You must write a name of file!", icon = "error", default = "ok")
    else{
      rbVal <- as.integer(tclvalue(dist))
      gt <- get("graphic.type", envir = envir)
      switch(rbVal,
             {if(gt == 1){
               pdf(paste(name, "OriginalCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "OriginalCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "OriginalRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "OriginalRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "OriginalMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "OriginalMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "CorrectedCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "CorrectedCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "CorrectedRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "CorrectedRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "CorrectedMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "CorrectedMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "NormalCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NormalCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "NormalRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NormalRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "NormalMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NormalMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "FilterCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "FilterCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "FilterRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "FilterRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "FilterMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "FilterMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "NoDuplicatesCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NoDuplicatesCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "NoDuplicatesRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NoDuplicatesRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "NoDuplicatesMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NoDuplicatesMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())})
    }
  })
  tkadd(optionsMenu,"command",label="Notes",command = function() note(envir))
  tkadd(topMenu,"cascade",label="Options",menu =optionsMenu)
    
  
  helpMenu <- tkmenu(topMenu,tearoff=FALSE)
  tkadd(helpMenu,"command",label="About genArise...", command=function() help())
  tkadd(topMenu,"cascade",label="Help",menu = helpMenu)

  sub.menu.frame <- tkframe(tt, relief = "groove", border = "2")
  
  image.menu.frame <- tkframe(sub.menu.frame, relief = "groove", border = "2")
  
  tcl("image","create","photo","filter",file=file.path(get("icons.dir", envir = envir),"icons/filter.png"))
  filter.button <- tkbutton(image.menu.frame,image="filter", command = function(){filter.gui(txt, img)})
  tkconfigure(filter.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","normal",file=file.path(get("icons.dir", envir = envir),"icons/norm.png"))
  normal.button <- tkbutton(image.menu.frame,image="normal", command = function(){normalized.gui(1, txt, img)
                                                                                if(swap==1){
                                                                                  assign("norm.alg", 1, envir = envir)}
                                                                                }) 
  tkconfigure(normal.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","zscore",file=file.path(get("icons.dir", envir = envir),"icons/zscore.png"))
  zscore.button <- tkbutton(image.menu.frame,image="zscore", command = function() zscore.gui())
  tkconfigure(zscore.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","cys",file=file.path(get("icons.dir", envir = envir),"icons/cy3vscy5.png"))
  cys.button <- tkbutton(image.menu.frame,image="cys", command = function() cys.plot())
  tkconfigure(cys.button, heigh = 16, width = 16)

  tcl("image","create","photo","acroread",file=file.path(get("icons.dir", envir = envir),"icons/acroread.png"))
  acroread.button <- tkbutton(image.menu.frame,image="acroread", command = function(){
    name <- tclvalue(tkgetSaveFile(initialdir = get("path.graphics", envir = envir),
                                   initialfile=get("spot.name",envir=envir),
                                   filetypes="{{PDF Files} {.pdf}} {{All files} *}"))
    name <- unlist(strsplit(name, "\\."))[1]
    if (!nchar(name))
      tkmessageBox(parent = tt,  message= "You must write a name of file!", icon = "error", default = "ok")
    else{
      rbVal <- as.integer(tclvalue(dist))
      gt <- get("graphic.type", envir = envir)
      switch(rbVal,
             {if(gt == 1){
               pdf(paste(name, "OriginalCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "OriginalCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "OriginalRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "OriginalRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "OriginalMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "OriginalMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "CorrectedCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "CorrectedCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "CorrectedRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "CorrectedRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "CorrectedMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "CorrectedMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "NormalCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NormalCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "NormalRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NormalRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "NormalMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NormalMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "FilterCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "FilterCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "FilterRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "FilterRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "FilterMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "FilterMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())},
             {if(gt == 1){
               pdf(paste(name, "NoDuplicatesCy3vsCy5.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NoDuplicatesCy3vsCy5.pdf", sep = "_"))}
              else if(gt == 2){
                pdf(paste(name, "NoDuplicatesRvsI.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NoDuplicatesRvsI.pdf", sep = "_"))}
              else{
                pdf(paste(name, "NoDuplicatesMvsA.pdf", sep = "_"),horiz=F, height=8,width=8,title=paste(name, "NoDuplicatesMvsA.pdf", sep = "_"))};
              graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir))
              dev.off(dev.cur())})
    }
  }) 
  tkconfigure(acroread.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","editor",file=file.path(get("icons.dir", envir = envir),"icons/editor.png"))
  editor.button <- tkbutton(image.menu.frame,image="editor", command = function() note(envir))
  tkconfigure(editor.button, heigh = 16, width = 16)
  
  tcl("image","create","photo","help",file=file.path(get("icons.dir", envir = envir),"icons/help.png"))
  help.button <- tkbutton(image.menu.frame,image="help",command = function() help())
  tkconfigure(help.button, heigh = 16, width = 16)

  tcl("image","create","photo","back",file=file.path(get("icons.dir", envir = envir),"icons/regresar.png"))
  back.button <- tkbutton(image.menu.frame,image="back",command =  function(){
    sure <- back.gui(envir)
    if(sure == 0){
      tkdestroy(tt)
      principal(envir,0)
    }
  } )
  tkconfigure(back.button, heigh = 16, width = 16)
  
  etiqueta <- tklabel(sub.menu.frame, text = "")
  tkconfigure(etiqueta, text = "", width = "80") 
  
  
  tkgrid(filter.button, normal.button, zscore.button, cys.button, acroread.button, editor.button, back.button, help.button, sticky = "nw")
  tkgrid(image.menu.frame, etiqueta, sticky = "w")
  tkgrid(sub.menu.frame)
  
  label.function <-  function(texto.etiqueta) tkconfigure(etiqueta, text = texto.etiqueta, width = "80")
  
  
  tkbind(filter.button, "<Enter>", function() label.function("Filter spots by intensity"))
  tkbind(filter.button, "<Leave>", function() label.function(""))
  
  tkbind(normal.button, "<Enter>", function()  label.function("Grid normalized spot"))
  tkbind(normal.button, "<Leave>", function() label.function(""))
  
  tkbind(zscore.button, "<Enter>", function() label.function("Obtain the z-score value"))
  tkbind(zscore.button, "<Leave>", function() label.function(""))

  tkbind(cys.button, "<Enter>", function() label.function("Plot log(Cy5) vs log(Cy3) values"))
  tkbind(cys.button, "<Leave>", function() label.function(""))
  
  tkbind(acroread.button, "<Enter>", function() label.function("Save graphic as PDF"))
  tkbind(acroread.button, "<Leave>", function() label.function(""))
  
  tkbind(editor.button, "<Enter>", function() label.function("Notes about this experiment"))
  tkbind(editor.button, "<Leave>", function() label.function(""))
 
  tkbind(back.button, "<Enter>", function() label.function("Back to the last window"))
  tkbind(back.button, "<Leave>", function() label.function(""))
  
  tkbind(help.button, "<Enter>", function() label.function("Help about genArise"))
  tkbind(help.button, "<Leave>", function() label.function(""))

  
  
  frame.label <- tkframe(tt,relief="groove",borderwidth=2)
  tkfocus(tt)
  assign("a.spot", get("o.spot", envir = envir), envir = envir)
  img <- tkrplot(upper.frame,fun = function()graphic.choose(get("a.spot", envir = envir), get("graphic.type", envir = envir)), hscale= get("Myhscale", envir = envir),vscale = get("Myvscale",
                                                                                                                                                                        envir = envir))
  tkgrid(img,frame1,padx="10",pady="10")
  tkgrid(upper.frame, pady = "10")
  
  area.frame <- tkframe(tt,relief="groove")
  yscr <- tkscrollbar(area.frame, repeatinterval = 5, command=function(...)tkyview(txt,...))
  txt <- tktext(area.frame,bg="white",font="courier",width =get("barWidth",envir=envir),height=get("barHeight",envir=envir),yscrollcommand=function(...)tkset(yscr,...))
  tkgrid(txt, yscr)
  tkgrid.configure(yscr,rowspan=4,sticky="nsw")
  tkinsert(txt,"end",texto)
  tkconfigure(txt, state="disabled")
  tkgrid(area.frame, padx = "10", pady = "10", sticky = "ew")
  if(follow.wizard){
    if(swap==0){
      op.counter <<- op.counter + 1
      write.spot(get("o.spot", envir = envir), paste(get("path.results", envir = envir),
                                 .Platform$file.sep, "original.txt",sep = ""))
      set.history.project(get("history.project", envir = envir), "Original Spot", "original.txt")
      c.spot <- bg.correct(get("o.spot", envir = envir))
      op.counter <<- op.counter + 1
      tkconfigure(txt, state="normal")
      tkinsert(txt,"end","\n\nBackground correction done.............")
      tkconfigure(txt, state="disable")
      assign("c.spot", c.spot, envir = envir)
      write.spot(get("c.spot", envir = envir), paste(get("path.results", envir = envir),
                                 .Platform$file.sep, "corrected.txt",sep = ""))
      set.history.project(get("history.project", envir = envir), "Corrected Spot", "corrected.txt")
      assign("a.spot", c.spot, envir = envir)
      tkpack(corr.radio,anchor="w",padx="10")
      tkselect(corr.radio)
      assign("corrected",1, envir = envir)
      normalized.gui(1, txt, img)
      filter.gui(txt, img)   #disabled temp
      remove.duplicates.gui(3, txt, img)
    }
    else if(swap==1){
      op <- get("op.counter", envir = envir) + 1
      assign("op.counter", op, envir = envir)
      write.spot(get("o.spot", envir = envir), paste(get("path.results", envir = envir),
                                 .Platform$file.sep, "original.txt",sep = ""))
          set.history.project(get("history.project", envir = envir), "Original Spot", "original.txt")
      normalized.gui(1, txt, img)
      assign("norm.alg", 1, envir = envir)
      filter.gui(txt, img)   #disabled temp
      remove.duplicates.gui(3, txt, img)
      assign("unique.alg", 3, envir = envir)
    }
  }
}