I am studying the cell and chromosome of Trichogramma(Hym.: Trichogrammatidae). I used of colchicinfor the cessation of cell division and acetic- orcein for staining of them. I got the images of this process. is the images shows the cells?
I think the images show cells, many of them in the metafase stage of mitosis.
That is what usually happens when you use colchicine to stop mitosis.
The figures are not clear enough to give a more detailed answer. But if you can send figures with a little stronger enlargement, may be we can go further in this preliminary opinion.
The quality of the photographs is poor so that I am uncertain what the photographs are showing. I suppose that you squashed the cells by pressing them. Thus, you do not expect to see the membrane/limits between cells, but some cell elements and also the exploded chromosomic plates. It is recommended the use a phase contrast microscope to observe the chromosomes under the stain that you have used. Please, make sure that you are using the appropriate microscope, the right contrast and the correct section in the squash. It takes a long time, sometimes, to survey the whole preparation, just searching for the correct spots with isolated metaphase chromosomic plates.
I agree you would have to have better quality images to make much of a determination but have you tried alternative protocols? or has this one worked well in the past? for instance if I was doing this on human cells, I would collect the cells, pellet them, expose them to a mild hypotonic solution (we use 0.075M KCl + 5%FBS) for 10-15 minutes, then fix the pellet in 3:1 methanol:acetic acid. Cells are affixed to slides with this fixative as well and then stained with Wrights (or Giemsa) stain for Brightfield microscopy.
Dear Gloria
I used other microscope, contrast microscope,, and send the new images but I did not able to see any chromosome. what is your solution?
Dear Nichole
when I used fixative, my photos did not clearly insufficient so again I could not see Chromosome. I fix them by absolute methanol: acetic acid(3:1).
Dear Fatemeh,
I have checked the new photographs that you uploaded and I do not see chromosomes. Considering that you squashed the cells, and the results that you are showing, I have the impression that the pressure that you put on the squash was not enough to display the metaphasic plates. I would suggest that you repeat again the whole process, being very careful in the times used for each steps of the protocol. Then, when you are at the point to squash your preparation, put as much pressure as you can, in an even way. It is not only to follow the steps of the preparation procedure, you have to learn also how to press the preparation. Sometimes, it takes time to learn just how much pressure to apply. We learn this just trying again and again.
Hoe that this help.
pleases end to a parasitology lab I am not specialist of parasites
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