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In this video we’ll be preparing route tips of eliam species to observe cells in various stages of mitotic division mitosis is one of the stages in the cell cycle where the chromosome of a cell replicate and separate to eventually form two genetically identical cells and like in meiosis chromosome number is maintained in both dollars cells for the experiment you’ll
Need onion or garlic bulb beakers toothpicks car noise fluid seventy percent ethanol one normal hydrochloric acid acidic army or a zero or c in stained glass lights and coverslips blade or scalpel watch glasses or petri dishes tiny vials or containers spirit lamp or bunsen burner or any flame source blotting paper droppers thumb forceps and a pair of scissor compound
Light microscope and immersion oil to begin the experiment take an onion or garlic bulb and fix it on a beaker containing tap water using toothpicks like you see here make sure the base of the bulb touches the water level keep this in a safe corner for a couple of days once the roots have grown about two to three centimeters cut out one centimeter of root tips and
Transfer them into a tiny vial containing car noise fixative fluid car noise fluid fixes the dna of the root tip cells leave the roots in the fixative for about 48 hours if you’re planning on storing the root tips for a longer time for use in future you may transfer the fixed route tips in a second vial containing 70% ethanol ethanol dehydrates the root tissue and
Thereby preserved dna root tips preserved in ethanol and preferably refrigerated will keep well for a couple of years for genetic studies either ways take a few root tips from car noise fluid or ethanol and transfer them onto a watch glass containing one normal hydrochloric acid as it will soften the cell walls and weaken cellular connections so it becomes easy
To squash the root material later gently warm the watch glass on a flame for about five seconds expose the root tips in the acid for about two minutes give the root tips a couple of washings in distilled water now transfer the roots onto a watch glass containing a pseudo carmen or acid or seen stain these are excellent chromosomal stains and will impart a deep
Red coloration to the nuclear material of the root cells warm the stain on the flame for about five seconds and leave the root tips in the stain for about five to ten minutes now transfer the root tips onto a clean glass light containing a drop of water using a sharp blade or scalpel remove about a millimeter of the root tip and discard the rest the very tips of
The roots are the regions with active cell division and they are all that we require for mitotic experiments gently lower a coverslip on the root tips making sure to avoid any air bubbles using the blunt end of a forcep or a pencil gently tap the cover slip a few times until the root tips are uniformly squashed in between the slide and the cover slip a properly
Squashed slide will appear faint cloudy pink to almost colorless the root tips light is now ready for microscopic observation this is how a finely squashed root tip appears under 10x of the microscope as you can see the cells of the root tips are nicely spread out with almost no overlapping x’ if root tips are not squashed enough you may repeat the squashing
Process until you get well spread root cells scan the slide for cells showing mitotic division stages under low power and then proceed to observation under high power such as 40 x or 100x oil immersion lens for observations under 100x objective you’ll need to add a drop of immersion oil on top of the coverslip and gently lower the objective lens until it touches
The oil surface from there use the fine adjustment knob for focusing you may take micro photographs of the various stages by employing a simple technique using a mobile phone and tripod like so and here’s a quick slide show the various mitotic stages of cell division in route tip cells of eliam species you
Transcribed from video
Mitosis in Onion Root tip Experiment By ThomasTKtungnung