Prepare and stain a root tip squash to observe the stages of mitosis.
Equipment•Root tip•Sharp knife•Acetic alcohol•HCl acid•Ice cold distilled water•Water bath at 60̊C•Test tube•Microscope, slides•Mounted needle•Toluidine blue stain
Method1)Cut small piece of root tip2) The root tip needs to be macerated•Place test tube of HCl in water bath, leave to heat•Place root tip in alcohol for 10 mins•Remove tip, place in ice cold water for a few mins.•Place tip in heated HCl for a few mins.•Repeat this
3) Break the tip with the mounted needle4) Prepare slide, add dye (blot out excess dye) view at 100x - 400x magnification.5) Count number of cells in one frame. Count the number of cells with visible chromosomes, in the same frame.
CalculationCalculate the mitotic index:Mitotic index=Number of cells with chromosomes/Total number of cells