Eosin, also called erythrosin, is a xanthene dye among synthetic dyes. It is a peach-red or pink powder, and is most suitable for use in combination with hematoxylin to display the morphological structure of normal or pathological tissues.The Eosin Staining Solution (water-soluble, 0.5%) is prepared with imported eosin dye. It is simple to operate and does not use toxic reagents such as mercury or methanol. It can be used for staining tissue sections or cultured cells, and can be used in combination with immunofluorescence staining or immunohistochemical staining. On the one hand, immunofluorescence staining or staining with other dyes can be performed after eosin staining; on the other hand, eosin counterstaining can be performed after immunohistochemical staining. This staining solution can be reused until the effect is considered unsatisfactory, at which point a fresh staining solution should be used instead. After staining, the cytoplasm appears pink or red.
Instructions:
I. Sample Preparation
(I) Paraffin Section Processing
1. Bake the sections for 30-60 min, then deparaffinize with xylene (I) for 5-10 min.
2. Deparaffinize with xylene (II) for 5-10 min.
3. Wash off the xylene with anhydrous ethanol for 1-5 min.
4. Treat with 95% ethanol for 1-5 min.
5. Treat with 75% ethanol for 1-5 min.
6. Rinse with tap water or distilled water.
(II) Frozen Section Processing
1. Deparaffinization is not required. After fixation with the fixative, rinse directly with distilled water for 2-3 min.
(III) Cell Sample Processing
1. Fix with 4% paraformaldehyde for 10-20 min.
2. Rinse with tap water twice, 2 min each time.
3. Rinse with distilled water twice, 2 min each time.
II. Staining
(I) Stain with the Eosin Staining Solution for 2-5 min (adjust the time according to the staining results and requirements).
If direct observation is required, wash twice with 70% ethanol. If dehydration, clearing and mounting are required, proceed with the subsequent steps; after washing with 70% ethanol, the subsequent steps of dehydration, clearing and mounting can still be followed.
Note: If it is used for counterstaining after staining such as immunohistochemistry, refer to the above steps and perform eosin staining after the other staining has been completed.
III. Dehydration, Clearing and Mounting (Optional)
(I) Wash with 95% ethanol (I) for 0.5-2 min.
(II) Dehydrate with 95% ethanol (II) for 2-5 min.
(III) Dehydrate with anhydrous ethanol (I) for 2-5 min.
(IV) Dehydrate with anhydrous ethanol (II) for 2-5 min.
(V) Clear with xylene (I) for 1 min.
(VI) Clear with xylene (II) for 1 min, then mount with neutral resin or other mounting media.
(VII) Observe under a microscope; the cytoplasm appears pink or red.
IV. Performing Other Staining
(I) If immunofluorescence staining or staining with fluorescent dyes such as Hoechst is to be performed, wash twice with 70% ethanol after staining with the Eosin Staining Solution.
(II) Soak in a solution used for immunostaining or fluorescent dye staining, such as PBS, normal saline, TBS or TBST, for 5 min.
(III) Perform immunofluorescence staining or staining with other fluorescent dyes.
Precautions:
1. If dehydration, clearing and mounting are required, prepare xylene, neutral gum or other mounting media yourself. If the sample is a paraffin section, prepare 90% ethanol, anhydrous ethanol and xylene yourself.
2. For your safety and health, please wear a laboratory coat and disposable gloves when operating.
Staining results
ZJ25F0420608