Hi everyone ! Any tips and tricks to section 3D culture/ stem cell spheroid of 1K cells density using Histo gel or Agar gel ?
You can follow these questions, may get some info.
https://www.researchgate.net/post/How_can_I_make_sections_from_spheroids
https://www.researchgate.net/post/Could_you_suggest_any_spheroids_cryosectioning_protocol
https://www.researchgate.net/post/What_is_the_most_efficient_method_for_IHC_of_spheroid_organoids
Immunostaining of spheresAfter 14–21 days of in vitro culture, matrigel discs con-taining the spheres were embedded in Histogel and then inparaffin for sectioning. Sections were processed and immu-nofluorescence was performed as previously described [1].Primary antibodies used were Rabbit Keratin 5 (1:200; Cov-ance), Goat polymeric immunoglobulin receptor (pIgR)(1:100; R&D Systems), Rabbit ALDH1/2 (1:100; Santa Cruz),Rabbit ALDH1A1 and Rabbit ALDH2 (1:100; Abcam), RabbitALDH3A1 (1:50; Abgent), Rabbit Keratin 14 (1:200; Epi-tomics), and Rat Ki67 (1:100; Dako). To detect mucus secre-tions in spheres, Alcian blue and Periodic Acid Schiffstaining was performed, as previously described [1] (PDF) Aldehyde Dehydrogenase Activity Enriches for Proximal Airway Basal Stem Cells and Promotes Their Proliferation. Available from: https://www.researchgate.net/publication/258202137_Aldehyde_Dehydrogenase_Activity_Enriches_for_Proximal_Airway_Basal_Stem_Cells_and_Promotes_Their_Proliferation [accessed Mar 23 2020].
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