Lab 3 mitosis and meiosis Experiment 1: observation of mitosis in a plant cell I

Question

Lab 3 mitosis and meiosis Experiment 1: observation of mitosis in a plant cell In this experiment, we will look at the different stage of mitosis in an onion cell. Remember that mitosis only occupies one to two hours while interphase can take anywhere from 18-24 hours. Using this information and the data from your experiment, you csn estimate the percentage of cells in each stage of the cell cycle Lab 3 mitosis and meiosis Experiment 1: observation of mitosis in a plant cell In this experiment, we will look at the different stage of mitosis in an onion cell. Remember that mitosis only occupies one to two hours while interphase can take anywhere from 18-24 hours. Using this information and the data from your experiment, you csn estimate the percentage of cells in each stage of the cell cycle Experiment 1: observation of mitosis in a plant cell In this experiment, we will look at the different stage of mitosis in an onion cell. Remember that mitosis only occupies one to two hours while interphase can take anywhere from 18-24 hours. Using this information and the data from your experiment, you csn estimate the percentage of cells in each stage of the cell cycle

Explanation / Answer

There are two ways you can go about it:

(1) Calculating the Mitotic Index

When you make chromosomes over slide, you count the no.of plates and count the no.of intact cells that did not burst to form chromosome plate.
The Ratio of no.of plates to total no of cells gives you the MI i.e. mitotic index. More the delay in cell cycle lesser is the MI.

(2) BrdU Incorporation study

Induct BrDu during active cell division. It is easier to do it in blood or bone marrow culture.
After you chromosome plates, Stain the slides with Hoechst 33258 and then do the SSC (sodium saline citrate treatment) under the sun.
Finally stain with Giemsa to see the the following:

    (i) dark stained arms [first mitotic cycle]
    (ii) one dark & one light stained chromatid [second mitotic cycle]
    (iii) both chromatids lightly stained [third mitotic cycle]

from this data we calculate the AGT (Average generation time) of the cell.

(3) Advanced way

MArk with florescent reagents which are tagged with markers of different cell cycle viz M, G1, S & G2 markers which are antibodies to particualr proteins like cyclin -cdks
then do a FISH or Flowcytometry to distinguish populations in different cell cycle phases

Regards

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