Can 70% ethanol-stored stigmas be used for pollen tube visualization after long-term storage?

23 June 2025 0 3K Report

I'm conducting field research on pollen tube arrest dynamics in self- vs. cross-pollinated flowers. My protocol involves:

Hand-pollinating flowers in the field

Collecting stigmas at multiple time points (2h, 6h, 12h, 24h post-pollination)

Immediately storing samples in 70% ethanol in microcentrifuge tubes (according to Techniques for Pollination Biology, Kearns and Inouye)

Processing after ~1 month of field storage

I plan to use aniline blue staining to visualize pollen tube growth and arrest sites.

Will pollen tubes remain detectable after extended 70% ethanol storage?
Are there any proven protocols for aniline blue staining after ethanol fixation?

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