Recommendations for avoiding agglomerates with WGA/Alexa Fluor 555

Dear all,

we are working with the Cell Painting workflow described in Nature Protocols 2016 [1] and are having an issue with the Wheat-germ agglutinin/Alexa Fluor 555 conjugate dye. We prepare stock solutions in water, but despite centrifugation as recommended in the paper we see small particles of agglomerates in the images for that respective channel.
We tried filtration, but too much of the dye seemed to be adsorbed to the filter membrane and the staining was weak.
Does anyone have similar experience and maybe some recommendation on how to better handle this dye?

Many thanks in advance.

Kind regards,

1.Bray, M.-A. et al. Cell Painting, a high-content image-based assay for morphological profiling using multiplexed fluorescent dyes. Nature Protocols 11, 1757–1774 (2016).

I am replying ridiculously late, but we do have a section of the public Cell Painting wiki dedicated to WGA; does this help (or were you able to solve the problem)?

Hi Anne, thanks for coming back to this. We could resolve this issue of agglomerates by using a new batch of the reagent.
KR Axel

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Wow, great to know that is a possible fix! Thanks for sharing the solution to your mystery.