Fig 1.
Representative cytology presentations of metastatic carcinoma.
(A) stomach, (B) lung, (C) pancreatobiliary tract, (D) breast, (E) ovary and (F) colon cancer.
Table 1.
A summary of the key morphological features identified in metastatic carcinoma present within ascites cases.
Fig 2.
Optimizing the order of dual immunocytochemistry for CK7 and PAX8.
When the antibody that is used first affects the overall manifestation because the first applied color becomes weak or even completely lost following staining with the second color. (A and B) Nuclear staining, using PAX8-specific antibody with DAB, was performed first followed by cytoplasmic or membranous staining using CK7-specific antibody with Fast Red. (C and D) CK7 staining was performed first followed by PAX8 staining. The initially applied CK7 red color becomes completely lost following DAB staining in the same case as in A and B.
Fig 3.
Typical immunocytochemistry patterns of cluster in peritoneal suspensions using LBC.
(A) Two-dimensional sheet with a dual-negative (CK7-/PAX8-) pattern represents mesothelial cells. (B) Three-dimensional large clusters with dual staining for nuclear PAX8 and cytoplasmic CK7. (C) Three-dimensional large clusters with staining for cytoplasmic CK7, but no nuclear PAX8 staining. (D) Two-dimensional sheet with staining for CK7 indicates that the cells are not mesothelial cells.
Fig 4.
Typical immunocytochemistry patterns based on primary origins.
Three parameters, three-dimensional (3D) clusters and dual staining for CK7 and PAX8, can identify the primary origin. Each panel represents different patterns of dual staining. (A) Stomach cancer: most cells are scattered and show CK7+/PAX8- immunoprofile. (B) Gastrointestinal tract, including stomach and pancreatobiliary tract cancer: mostly small 3D cluster with scattered cells are CK7+/PAX8-. (C and D) Female genital tract, including ovarian cancer: large 3D cluster with scattered cells are CK7+/PAX8+. There is positive staining for nuclear PAX8-DAB in the cell with the large cytoplasm.
Table 2.
The summary of morphological and immunocytochemistry patterns.
Fig 5.
Automatic digitalized interpretation of dual immunocytochemistry.
In peritoneal aspirates from patients with ovarian cancer, the suspended cells were very heterogeneous: a mixed population of mesothelial cells and malignant tumor cells. Except for 265 mesothelial cells with dual negative staining (CK7-/PAX8-), all cells were compatible with malignant cells. Approximately 30% of the cells had the typical dual staining (CK7+/PAX8+) pattern.
Fig 6.
Automatic digitalized interpretation of dual immunocytochemistry.
In most cases of ovarian cancer, cells with the typical dual staining (CK7+/PAX8+) pattern represented over 90% of the cell population.
Fig 7.
Estimated ROC curve analysis using the three parameters to predict primary origin.
(A) In stomach cancer, AUC was 0.8699. (B) In ovarian cancer, the AUC was 0.9812. (C) In lung cancer, the AUC was 0.882. (D) In pancreatobiliary tract cancer, the AUC was 0.8772.
Fig 8.
Diagnostic decision tree for peritoneal fluid cytology utilizing cellular cluster and immunocytochemistry patterns to identify primary cancers which have metastatized to peritoneal fluids.