First Trial of Cytogenetic Test on Bone Marrow Sample

After coming back from Singapore, I have been given a precious bone marrow sample to do the cytogenetic test from Prof. Hany to be tested. This was the first time for me to deal with the bone marrow sample. With excitement, I started off with the culturing process.

Obstacles of Cytogenetic Test

However, there were many obstacles for me to do the test. The materials and reagents that I need for doing this test are not available at that time, especially for the essential culture medium. Consequently, I need to think of the alternative. I tried to use the transfer medium provided by National University Hospital Singapore to do the culturing process with the hope to grow at least some leukemic cells from the sample.

Furthermore, I do not have the pre-made ethidium bromide solution yet. The ethidium bromide solution is important to elongate the chromosomes. So, I have to make it myself by using the powder-formed ethidium bromide.

Outcome of Cytogenetic Test

After culturing and harvesting the bone marrow cells, I used the cell suspension obtained to prepare the slide. Then, I viewed the slide under the microscope. There were just few metaphases and the chromosomes not spread well. I am going to stain the chromosome on the next day. I hope that I can still get some good metaphases to be analyzed via imaging system.

Troubleshooting the Problems

In my opinion, this is not a good result that I want to get from the cytogenetic test. Why?

1. There were a lot of dead cells when I viewed the slides under microscope. This might probably because of the cells not grow well during the culturing process.

2. The length of the chromosomes was short. This indicated that the elongation step was not performed well.

3. The mitotic index was low. This might because of the metaphase arrest step was not performed well. Yet, the mitotic index is closely related to the condition of the sample and the patients too.

4. The chromosomes not spread well. This might because of the slide preparation step was not performed well. This step closely related to the technique used by the technologist. I think I am not doing well when performing this step.

Solutions for the Problems

To solve those problems, there are many optimization steps that I need to carry out.

1. I have to use the appropriate culture medium in order to make sure the cells can grow well especially for the leukemic cells. I am ordering the Chang Medium for bone marrow culture from Irvine Company.

2. The home-made ethidium bromide solution seems like not work so well. Therefore, I decided to order the pre-made ethidium bromide solution too to make sure the chromosomes can be elongated.

3. The amount of the reagents used should be accurate.

4. I will try my best to improve my technique when performing the slide preparation step to make sure the chromosomes can spread well on the slide.

Summing up, after the first trial, there are still plenty of things for me to do and learn. Tomorrow I will continue with the staining step and start to analyze the metaphases. Stay tuned in Cytogenetics Cancer Research blog to see what I will get from this first trial.