We are performing FISH in plasma cells. When it comes to back up, we are using a small amount of EDTA in order to store those cells. In case of a repeat, the EDTA creates a kind of membrane in the microscope slide that can not be removed very easily. Usually, for this purpose, I use some drops of Fixation buffer (Acetic acid/Methanol) but as a result I am loosing a lot of previous existing cells. Does anyone know of a proper way? Am I eligible to use 70% ethanol, without disrupting the FISH protocol at the same time?
EDTA is commonly used as an anticoagulant to preserve cells in blood samples, including plasma cells, for downstream analysis such as FISH. However, EDTA can form a complex with calcium ions, which can cause cells to adhere to the surface of a microscope slide and create a "membrane" that is difficult to remove. This can be a problem when repeating the FISH assay, as the cells may not be fully released from the slide and may be lost during the process.
One option for removing the EDTA-induced membrane from a microscope slide is to use a solution of acetic acid and methanol, which can dissolve the calcium-EDTA complex and allow the cells to be released. However, this solution can also cause the cells to be lost or damaged, as you have observed.
An alternative option is to use a solution of 70% ethanol to remove the EDTA membrane. Ethanol is less aggressive than acetic acid/methanol and may be less likely to damage the cells. To use 70% ethanol, you can place the slide in the solution for a few minutes and then gently rinse the slide with water to remove any residual ethanol. This should help to release the cells from the slide without damaging them.
It is worth noting that the optimal method for removing the EDTA membrane will depend on the specific requirements of the experiment and the characteristics of the samples being analyzed. It may be helpful to consult with experts or refer to published protocols for guidance.
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