I am currently planning to conduct ploidy analysis on plants using flow cytometry.
The original protocol suggests using fresh young leaves for the analysis, but I'm facing difficulties in maintaining the samples in a fresh state.
I'm wondering if you could provide some advice on suitable sample storage methods.
For bacterial cell counting, it seems effective to store samples in ethanol at -20C or in a deep freezer.
However, I'm curious if these storage methods would also be useful when measuring the genome size of plants.
Thank you for your assistance.
The key is minimizing enzymatic and metabolic activities that could degrade nuclei. Quick freezing or fixation right after collection provides the best preservation. Proper sample storage is crucial for getting accurate ploidy results.
try to:
- Freezing fresh leaves in liquid nitrogen or a -80C freezer immediately after collection can help preserve nuclei integrity. The frozen leaves can be stored this way for months before analysis.
- Fixing leaves in 3:1 ethanol:acetic acid solution is another common method. The fixed samples can be stored at 4C for a few days or at -20C for longer periods. Fixation helps stabilize nuclei.
- For short term storage (up to 24 hours), freshly collected leaves can be kept refrigerated in moist paper towels inside sealed plastic bags. This helps reduce metabolic activity.
- Drying samples rapidly using silica gel, then storing at room temperature can also work for short term storage. Rehydration is required before analysis.
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