Luna staining, also known as Luna-Bielschowsky staining or Luna staining for eosinophils, is a technique used to highlight eosinophils in histological specimens. This technique is useful to identify and quantify eosinophils in tissues, especially in studies of inflammation and allergic diseases. Here is a summary of the Luna staining technique we use:

Sample preparation: Prepare tissue samples following the appropriate fixation protocol. Usually, 10% neutral buffered formalin is used as the common fixative. Tissues are then dehydrated, embedded in paraffin, and cut into thin sections (approximately 4-6 μm) on slides.

Deparaffinization and rehydration: Immerse slides in xylene or a xylene substitute to remove paraffin, then rehydrate sections in a series of descending alcohol solutions (for example, 100%, 95%, and 70% ethyl alcohol) to remove paraffin. alcohol and rehydrate tissues.

Moon stain:

to. Eosinophils: Place sections in 0.5% eosin Y solution for approximately 10 minutes to stain eosinophils pink.

b. Methylene Blue: Next, dip the sections into a 1% methylene blue solution for a few seconds to contrast and highlight the eosinophils.

c. Dehydration: After methylene blue staining, dehydrate sections in a series of ascending alcohol solutions (eg, 70%, 95%, and 100% ethyl alcohol) to remove water and dehydrate tissues.

d. Rinsing and mounting: Then, rinse the sections in xylene or xylene substitute for rinsing, and finally mount the slides with a suitable mounting medium, such as a synthetic resin.

Observation: Sections stained with the Luna staining technique can be observed under a light microscope. Eosinophils will appear pink or red in color due to eosin, and will stand out in contrast to other tissue components.

Remember that Luna's staining technique can vary in details and staining times depending on the specific protocol used in your lab.

At the time that it is necessary and if it is required of you that it must necessarily be the Luna stain, I recommend using the Wright or Giemsa stain for tissues as a positive control, which will show the granules of a very evident red-orange color.

If you still have problems, do not hesitate to let me know to request more support from our technicians.

Greetings.

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