Hello,
I am looking for an acridine orange staining protocol for thick paraffin embedded plant rhizome sections. Also, any feedback on a suitable fixative would be appreciated. I have read that using acetic acid in the fixative is not advisable when using a fluorescent stain like acridine orange. Thank you.
Acridine Orange Protocol
Re: Acridine Orange Protocol
http://www.tandfonline.com/doi/abs/10.3 ... ode=ibih19
Stem, leaf, and bud tissue of sweet potato, tomato, and pepper were embedded in paraffin, sectioned, mounted, and stained with 0.01, 0.1 and 1% aqueous and 0.1% alcoholic solutions of acridine orange. Temporary and durable mounts were prepared and irradiated under short and long wave ultraviolet light. Intensity and specificity of the fluorescence imparted to tissues were chiefly affected by type of fixative. Best results were obtained with fixatives containing formalin but not acetic acid. Tests on the effect of pH obtained with McIlvaine's buffer between 4.5 and 8.3, and made only with the aqueous stain, showed 6–8 to be optimal. Aqueous staining 1 hr in 0.1% solution, pH 6–8 is recommended for temporary mounts. Durable mounts in a nonfluorescent resin can be made after differentiation in buffer and dehydration in dioxan solutions.
Stem, leaf, and bud tissue of sweet potato, tomato, and pepper were embedded in paraffin, sectioned, mounted, and stained with 0.01, 0.1 and 1% aqueous and 0.1% alcoholic solutions of acridine orange. Temporary and durable mounts were prepared and irradiated under short and long wave ultraviolet light. Intensity and specificity of the fluorescence imparted to tissues were chiefly affected by type of fixative. Best results were obtained with fixatives containing formalin but not acetic acid. Tests on the effect of pH obtained with McIlvaine's buffer between 4.5 and 8.3, and made only with the aqueous stain, showed 6–8 to be optimal. Aqueous staining 1 hr in 0.1% solution, pH 6–8 is recommended for temporary mounts. Durable mounts in a nonfluorescent resin can be made after differentiation in buffer and dehydration in dioxan solutions.
-Reichert Polyvar
-Olympus IX70
-Zeiss Photomicroscope
-Canon 600D
-Olympus IX70
-Zeiss Photomicroscope
-Canon 600D
-
microaussie
- Posts: 2
- Joined: Thu Apr 11, 2019 9:52 pm
Re: Acridine Orange Protocol
Hello Seb28,
Thanks for the reply, but I have already seen that. I would liked to have read more, but I am not willing to pay for it yet. I was hoping for a protocol laid out in more simple terms, as I am new to staining.
Thanks for the reply, but I have already seen that. I would liked to have read more, but I am not willing to pay for it yet. I was hoping for a protocol laid out in more simple terms, as I am new to staining.