Which is the best staining method or technique for counting chromosomes?
When I was trying to observe the chromosomes in the microscope (10*100X) eyepiece and objective lens, I could see the metaphase and late anaphase or telophase chromosomes but unable to count them... Could anyone please suggest the best method to count the individual chromosomes..?
Material: Onion root tip
The chromosones are very dwarf. So you need reducing concentration of proliferation suppressed factor.
You have not mentioned the material and the protocol you have used here. You need to manipulate the protocol. Also, the number of divisions is far too less. You might also want to check the time of fixation. You can message me if you need any further help.
yes you might also want to check the time of fixation and used colchicine solution for pretreatment for root tips
Counting of mitotic chromosomes is easier and faster. Root tips are the most convenient source of mitotic cells. When roots are not available, young buds, leaves or callus can be used. Cytological procedures of chromosome preparation and staining are modified depending upon plant species.
A year of bitter, expensive experience has taught me that prior to selecting microscope protocol you need to find out the approximate physical length of the chromosome in the species you are looking for. That measurement should be in um, not bp.
Monocot species like garlic are most often cited in online beginner tutorials. However their chromosome size is 10-15x larger than a lot of other plant species
If your chromosome size is smaller than about 10um you're unlikely to be able to visualise chromosomes easily with a standard biological microscope at 100x and a 10x objective unless the microscope is really good quality, you have good quality starting material and your protocol is refined to suit
For physically smaller chromosomes like Brassica you'll need to be very lucky, have a lot of time to spend or to have access to a fluorescence microscope
( Hoping someone can tell me I'm wrong )
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