ABC

After acridine orange staining, images are typically taken with the following filters:

• Green filter (FITC filter): For observing cytoplasmic staining, as acridine orange fluoresces green in the cytoplasm where it binds to nucleic acids.
• Red filter (Texas Red filter): For visualizing the acidic compartments (lysosomes), as acridine orange emits red fluorescence in these regions due to its protonation in acidic environments.

Using these filters will allow you to differentiate between intact lysosomes (red fluorescence) and areas where lysosomal integrity may be compromised (green fluorescence, indicating leakage).

1. • ombine the Giemsa stain and diluent in a glass container or staining jar.
   • Mix gently by swirling or using a vortex mixer to ensure a homogeneous solution.

2. Using the Stain:

   • Apply the diluted Giemsa solution to your samples as needed (e.g., for staining cells in an invasion assay).
   • Stain for the recommended time (usually 20-30 minutes).
   • Rinse gently with distilled water to remove excess stain.

3. Storage:

   • The working solution of Giemsa stain should ideally be prepared fresh and used immediately for best results. If necessary, it can be stored in a dark bottle at 4°C for a limited time (typically no more than a week) but should be discarded if any precipitate forms or if it changes color.

Summary

When starting with Giemsa stain in solution, you can easily dilute it to achieve the desired concentration for your specific application, ensuring proper staining and visualization of your samples