Hello All,
What is the expected spread amount on a single glass slide for a typical karyotyping procedure? Also, I know that a lot of the time it is hard to trace if all the chromosomes came from a single cell. So, how many of these spreads are usually useful.
I understand karyotyping is like an "art", so sometimes the process can be very subjective and hard to give an answer. However, I am just looking for an approximate number from someone that has experience with the process and has already done it many times.
Thank you so much!
Hi Reha,
In cell culture, if the cells have proliferated well and you have prepared a good slide, you can capture between 50-100 karyotyping areas. On the slide, the chromosomes for a single cell are usually close together within a certain area.
You are right about your observations regarding the realities of karyotyping. Very often the nuclear membrane of your 'fruitful' cell is torn! It so happens that the chromosomal elements are still clearly identifiable by their sizes and level of staining for cells in this category. You need to establish the chromosome number before capturing an image.
You are right that karyotyping is an art, particularly when you are studying chromosomes in the solid state. It is less so when they are banded. The morphological classes A to G must be mastered and as you harvest each image, you attempt to arrange them by the groups. I have found the knife tool on Corel Draw very useful but before this, karyograms were constructed using the blade on each chromosome image.
Mercifully, softwares now do all the sorting out and matching. For those of us who came from the analogue tradition, these images are etched somewhere in our consciousness!
Thank you so much for your insight and useful tips! I appreciate you take your time to respond to this.
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