Practice Guidelines

IHC and DIF are immunologic-based technologies using specific antibodies with different detection systems. The reaction product generates a colored reaction product which is evaluated under the standard light microscope (IHC) or fluorescence microscope (DIF). At Vitro, the use of DIF is largely restricted to fresh, skin biopsies for the evaluation of cutaneous-based immunologic conditions. The applications of IHC are innumerable, which makes this assay routinely used for the diagnostic and predictive markers workup in anatomic pathology. As an international anatomic pathology reference laboratory, Vitro has validated hundreds of (over 250) IHC antibodies that are currently being used in diagnostic workups. Antibodies List

While FISH was first introduced to the clinical testing arena in the early 90s, it now has become an essential component of the tools employed in the pathology laboratory. FISH applications now include a wide range, either as a diagnostic test helping confirm the diagnosis of a hematolymphoid tumor or a soft-tissue tumor, or as a prognostic/predictive companion diagnostic assay. Most clinical FISH probes include ‘direct labeling’ of specific nucleic acid sequences (gene specific, or chromosome specific) attached to fluorescent molecules. Dr. Yaziji and our pathologists are thought leaders in FISH testing, and have been extensively involved in clinical research on FISH assays since the nineties, and have personally reviewed and reported over ten thousands FISH assays to the pathology and oncology community throughout the United States, Asia and South America. Dr. Yaziji is also the author of a book (Springer, in press) entitled “FISH Testing in The Pathology Laboratory). Probes List

Chromogenic in situ hybridization (CISH) CISH utilizes similar detection systems as immunohistochemistry, but by hybridizing the nucleotide probe to the complementary sequence of interest, CISH can identify the presence of specific genes or chromosome segments in various neoplastic and non- neoplastic tissues. Vitro offers CISH-based testing of EBV mRNA, kappa light chain gene and lambda light chain gene. TESTS EBER1 - ISH Kappa (ISH) Lambda (ISH)

Flow cytometry is a technique for detecting and quantitating antigen expression in single cell suspensions. Fluorescein-conjugated antibody ‘panels’ are added to the cell suspension. The mixture of cells and bound antibodies will pass through the flow cytometer, and the cell-associated fluorescence in response to illumination with one or more lasers is recorded. Computer analysis of several patterns of protein expression can identify the immunophenotype of the cells of interest, thus helping to confirm a diagnosis of a specific hematolymphoid disorder, confirm light-chain restriction (which is a hallmark of neoplastic lymphoproliferative disorder), or detect a small residual population of malignant cells, following treatment. While flow cytometry has historically been used to characterize hematolymphoid cell populations, this technique can be applied to any monodispersed cell suspension. At Vitro Laboratories, cell-associated fluorescence is measured on a state-of-the-art, 3-laser/9-color Becton Dickinson LSRII flow cytometer, utilizing a collection of nearly 100 fluorescently-labeled antibodies. Flow Cytometry Panels

Polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA or RNA, which generates sufficient DNA for detection. Most, but not all molecular assays utilize PCR, which is often more sensitive than other molecular methods. Vitro uses PCR-based testing for B- and T-cell clonality testing in lymphoid lesions, and for allele-specific PCR testing of prognostic / predictive markers in oncology, such as KRAS, BRAF, and EGFR. Example of hematology-based assays offered at Vitro include MPL and JAK2. Molecular list

While these technology has been employed over a century ago, some technical advances have been made with regard to cytology monolayer processing and tissue processing. Routine cytopathology and histopathology remains the only gold standard technology used to process patients samples (such as pap smears, cervical biopsies, endometrial biopsies, prostate biopsies, skin biopsies, breast biopsies and GI biopsies) and render pathologic diagnoses on these samples. Special Stains