EA 50 reagent is a light green, alcohol solution of two acid dyes: Eosin Y and Light Green SF mixed in Phosphotungstic acid (PTA). It is used in Papanicolaou staining procedure for routine cytoplasmic staining to provide good color contrast and reproducible results that aid in the classification and identification of exfoliative cells.
EA 50 reagent is a light green, alcohol solution of two acid dyes: Eosin Y and Light Green SF mixed in Phosphotungstic acid (PTA). It is used in Papanicolaou staining procedure for routine cytoplasmic staining to provide good color contrast and reproducible results that aid in the classification and identification of exfoliative cells. The polychromatic EA solution stains the unstained cellular components, such as nucleoli, squamous cells, cilia, and erythrocytes. The test samples for Papanicolaou staining could be gynecological or non-gynecological including sputum, urine, and cytological samples. EA 50 reacts with the cytoplasm of the less mature squamous cell (exocervical), glandular cell (endocervical), and the basal, paranasal, and intermediate cells.
The Papanicolaou test is a staining procedure used to detect malignant cells in a sample of superficial cells that line the inner wall of the uterine cervix for early cancer diagnosis. The Pap smear is used for cheap and rapid examination of the gynecological and non-gynecological samples.
Clear, may have fine particles.
Green-colored solution with a reddish-purple tinge.
The formula of the staining solution can be adjusted and standardized as per the performance parameters.
The differentiation of cells is vital for the proper identification of the cell types found in the cytological smears.
Since there are many nuclear abnormalities in cancerous cells, the staining of the nucleus is important for an accurate diagnosis.
Transparency of the cytoplasm is of particular importance because of the frequent overlapping of cells and their varying thickness.
The Papanicolaou stain is a mixture of acidic and basic dyes. The acidic part stains the basic components of the cell and the basic stains the acidic components of the cell. The hematoxylin solution contains the nuclear stain which stains the cell nuclei blue. It has an affinity for chromatin particularly the sulfate groups on the DNA molecule. The Orange green 6 is the acidic counterstain which stains the matured and keratinized cells. The Eosin Azure (EA 36, EA 50, and EA 65) is a polychrome mixture of eosin Y, light green SF, and Bismarck brown. Eosin Y gives pinkish shade to the cytoplasm of mature squamous cells, nucleoli, cilia, and red blood cells. Bismarck brown Y stains nothing and is often omitted in cytological staining.
Fix the sample smears in 95% alcohol for 5-15 minutes.
Note: The quality of the staining depends on the fixative, age of the staining solution, thickness of the sample, temperature and pH of the water used, rinsing, and air drying.
Following results are obtained with EA 50 staining;
The EA 50 stain is used in the Papanicolaou staining technique that displays a characteristic range of coloration to exfoliative cells of cervical, vaginal, prostatic and other body secretions facilitating the examination of nuclei and other cytoplasmic components.
Evaluation of bladder neoplasms (Murphy, 1990)
The pathological examination of urinary specimens is vital for the detection and monitoring of the bladder neoplasms. Urinary cytology is one of the diagnostic techniques used for the early diagnosis of urinary neoplasms. The Papanicolaou stain mainly EA 50 is a powerful dye to decipher the cytological details of the urinary sample. The differential diagnosis of urothelial neoplasia versus the reactive/reparative changes secondary to urinary stones could be accomplished. The Papanicolaou staining has rendered new avenues in urinary neoplasia detection, staging, and treatment monitoring.
Eosin Azure 50 is a stable and convenient dye that reacts exclusively with the target cells aiding in their identification and accurate diagnosis. It enables the visualization of nuclear details, keratinized cells, and cytoplasmic machinery. For optimal staining results, the EA 50 stain should have properties that are in complete compliance with other reagents used in the Papanicolaou staining process.