Hello, I am a student attempting Ploidy analysis using a flow cytometry on plant samples.

I conducted experiments using FACS Aria II, but obtained results different from existing literature.

When gating the debris and examining fluorescence peaks, most values were clustered near 0 on a linear scale, making it difficult to confirm.

On a log scale, I obtained the following peaks; however, the peaks were formed at the same position for both 2n and 4n samples.

I couldn't find any papers performing similar analysis on FACS Aria II.

Could it be that this instrument is not suitable for ploidy analysis?

Experimental conditions are as follows:

Reagent: Used CyStain UV Precise P (performed staining with DAPI) following the manual(1 min staining time. Is it too short?).

Samples: Used 4n potato (C=1.82 pg) as an internal standard. 2n samples (in the case of closely related species, C=ca. 8 pg), 4n samples (in the case of closely related species, C=ca. 15 pg). Samples were stored at -20 in ethanol.

Thank you in advance for your assistance.