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Objectives
- Attain competency in interpreting flow cytometric histograms, dot plots, and contour plots.
- Understand the utility as well as the limits of the method and how the data generated integrates into the diagnostic interpretation.
- Acquire familiarity with criteria for diagnosis of malignant lymphoma using flow cytometry.
- Acquire familiarity with criteria for diagnosis of aneuploidy using DNA content analysis.
- Understand concept of determination of S phase fractions and variables important in this determination.
- Understand the need for quality control and know what currently is acceptable in this area.
Residents spend 1 month rotating on the Clinical Flow Cytometry Service as an elective rotation. In the first week the residents spend the majority of their time reading literature provided and discussing antigens and antibodies. During this time they are introduced to interpretation of patient immunophenotypic data and are guided through analyses of classical flow cytometric cases. As the residents acquire greater knowledge, they are given cases to review on their own for presentation at sign out. Reports are initially dictated to the residents but as they become more knowledgeable, they are required to prepare reports prior to review and sign out. These reports are then reviewed and corrected.
The Flow Cytometry Laboratory tests for the presence of hematopoietic neoplasms in specimens submitted (including blood, bone marrow, cell suspensions, fluids, fine needle aspirates, CSF). The Flow Cytometry Laboratory does not perform the following analyses: transplant patients- immune monitoring, immunodeficiency work-ups (immunophenotyping and/or CD4/CD8 counts), serologic tests for infectious disease, serologic autoimmune work-ups, other immunologic or histocompatibility testing procedures. Residents review the immunophenotypic data generated and compare to morphological findings before signing out cases with a staff member and then generating a report. Approximately 88 cases are reviewed in a one-month rotation. Residents attend conferences in which diagnostic molecular findings are compared to immunophenotypic data. Outside consult cases are reviewed with residents. Residents receive lectures on flow cytometric immunophenotyping.
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B. Hematopathology Rotation
Mentor
Dr. Elaine Jaffe
Duration
All residents complete a 1-month rotation in hematopathology. Additional elective rotations may be completed at the residents descretion.
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C. Immunopathology of the Lymphoid System
Mentors
- Dr. Elaine Jaffe
- Dr. Maryalice Stetler-Stevenson
During this immunopathology elective the resident will learn the role of immunologic techniques, including flow cytometry and immunohistochemistry, in the diagnosis of benign and malignant lymphoid lesions. The resident assumes responsibility for ordering and interpreting immunologic assays. The routine histopathologic findings are correlated with the results of frozen section immunohistochemistry and flow cytometry using a large battery of monoclonal antibodies. The resident will gain familiarity with the CD nomenclature for monoclonal antibodies applicable to the hematopoietic system. After reviewing the data with the staff pathologists, the resident will prepare an immunopathology report, which is issued as a supplement to the routine surgical pathology report.
Immunopathology training is also an integral part of the required hematopathology rotation supervised by Dr. Elaine Jaffe and the staff of the Hematopathology Section. During this elective the resident learns the role of immunologic techniques, including flow cytometry and immunohistochemistry, in the diagnosis of benign and malignant lymphoid lesions. The resident assumes the responsibility for ordering and interpreting immunologic assays. The routine histopathology findings are correlated with the results of immunohistochemistry and flow cytometry using a large battery of monoclonal antibodies. The resident gains familiarity with the CD nomenclature for lymphoid surface antigens. After reviewing data with the staff pathologists, the residents prepare an immunopathology report, which may be issued as a supplement to the routine surgical pathology report.
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D. Molecular Pathology Rotation
Mentors
Dr. Mark Raffeld
Overall goals and training
- The overall goal of the Molecular Pathology Section is to teach residents and fellows how to safely handle tissue samples for molecular analysis, to learn basic techniques at the laboratory bench, and to learn how to analyze results critically, including an appreciation of the limitations of each technique.
- Training is provided by didactic lecture, by hands-on bench experience, and in data review conferences.
Specific Training Objectives
- Learn basic techniques of nucleic acid and protein handling, including proper storage of tissue samples and the extraction of intact materials from tissue samples.
- Learn electrophoresis methods for analysis of DNA, RNA, and protein, including agarose gel electrophoresis, acrylamide gel electrophoresis, and denaturing agarose gel electrophoresis.
- Learn standard enzymatic treatments of DNA, including restriction enzyme analysis.
- Learn basic hybridization techniques such as Southern and Northern blot hybridization. Learn how to analyze restriction patterns of DNA to detect restriction fragment length polymorphisms (RFLPs), gene deletions, and gene amplifications. Learn how these molecular techniques complement classic cytogenetic methods.
- Learn basic polymerase chain reaction (PCR) techniques to amplify targeted DNA from tissue samples. Learn how to select primers and proper controls.
- Learn how to grow bacteria carrying recombinant plasmids, to isolate recombinant plasmids, and to purify cDNA inserts as molecular probes for hybridization analysis.
- Learn how to label probes with nonisotopic and isotopic methods. (All laboratory staff and visitors must pass a 1-day Radiation Safety course before using radioisotopes.)
Optional objectives
- Learn basic methods of in situ hybridization of cytospin and tissue samples.
- Learn basic molecular cloning techniques and screening of libraries.
- Learn basic DNA sequencing techniques.
For more information on clinical departments (e.g., procedures, staff information, specimen collection, forms, etc), please see:
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E. Molecular Diagnostics in Hematology Rotation
Mentor
Dr. Mark Raffeld
The Hematopathology Section of the Laboratory of Pathology, under the direction of Dr. Mark Raffeld, conducts molecular genetic analyses of patient lymphomas and leukemias. Samples are received directly from patients treated under National Cancer Institute protocols and processed for DNA extraction and gene rearrangement analysis.
Residents have the opportunity to observe and participate in the procedures undertaken, which include extraction of high molecular weight DNA, restriction endonuclease digestion, gel electrophoresis, Southern filter transfer, DNA-DNA hybridization, DNA probe labeling, autoradiography, and interpretation of data. Probes applied in these studies include those that are capable of detecting chromosomal abnormalities such as the translocations commonly seen in lymphoma and leukemia. By diagnosing a chromosomal translocation at the molecular (DNA) level, the resident under the guidance of the laboratory supervisor interprets the molecular evidence in light of conventional cytogenetic results.
Molecular cytogenetics performed in this way reveals chromosomal translocations in the majority of follicular lymphomas, the most common lymphoma in the United States.
This approach represents the most readily applied technique for detection of this common chromosomal abnormality, which involves the t(14;18) translocation, joining bcl-2 with the immunoglobulin heavy chain gene. In addition, the same methodology is used to detect the Philadelphia chromosome of chronic myelogenous leukemia, the chromosomal translocation associated with the c-myc proto-oncogene of Burkitt's lymphoma, and other examples of hematopoietic neoplasms. These studies provide pathology residents with insights into cytogenetics, molecular genetics, and pathogenesis of human neoplasia
For more information on clinical departments (e.g., procedures, staff information, specimen collection, forms, etc), please see:
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F. Ultrastructural Pathology Section Rotation
Mentor
Dr. Maria G. Tsokos
Duration
Integrated with Surgical Pathology and available as a separate 1-month elective
Objective
The objective of the Ultrastructural Pathology (UP) Section is to provide all residents with knowledge of the ultrastructure of pathologic conditions and with the understanding that the final diagnosis is the result of a combination of available methods. Therefore, the ultrastructural data are always evaluated in conjunction with the light microscopic appearance, the immunohistochemical data (when available), and the clinical information. This approach requires close interaction of the pathologist who is responsible for the electron microscopic (EM) diagnosis and the residents. The goal of the UP is to familiarize residents with the ultrastructural features of pathological conditions and to teach them how to use these features in combination with other available diagnostic tests and clinical information to reach a final diagnosis.
Training
- Pathology residents participate in the interpretive review process of the diagnostic EM cases during the time of their surgical rotations. They review and discuss the electron micrographs with the EM attending pathologist or fellow. The fellow or the attending issues EM reports. Research specimens are not evaluated by the residents, but by the EM fellow or the attending.
- All residents receive 2 EM lectures per year on the use of electron microscopy in the diagnosis of tumor and other diseases. Material from both diagnostic and research specimens is used as teaching material in these lectures.
- Residents rotating through the UP laboratory receive hands on training on the practical and theoretical aspects of diagnostic EM. Specifically, they become familiar with the steps involved in the preparation of specimens for EM examination from embedding to cutting to staining thick and thin sections. They are also taught about proper fixatives used for electron microscopic evaluation of a specimen. At the beginning of the rotation, a medical technologist shows the resident how to use the electron microscope to scope and photograph pertinent ultrastructural features. The resident is given a set of 15 teaching cases that have been carefully selected by the director of the section to cover appropriate topics in the field of diagnostic EM. Short clinical summaries, H+E slides, and thick and thin sections accompany each case of the set. The resident is asked to use the available information and to photograph ultrastructural fields that will lead him/her to a diagnosis. The resident meets with the director of the section to discuss the EM findings and obtain pertinent literature and advice. At the end of the rotation, the resident is able to handle the basics of a diagnostic EM case and to proceed with the formulation of an EM report. Specifically, in the last week of the rotation, the resident is instructed how to present the ultrastructural findings and how to correlate these findings with other pertinent clinical information.
- Teaching on Individual Basis: All diagnostic EM cases are reviewed by the pathologist in the UP section and the resident who signs out the corresponding surgical pathology report. This interaction provides most of the individual EM teaching to our residents. The EM findings are included either in the surgical pathology report, signed by the resident and the staff pathologist, or in a supplemental EM report.
Theoretical Teaching/Lectures
All residents receive additional theoretical EM teaching in lectures offered by Dr. Maria Tsokos as part of the scheduled teaching conferences for the residents by the Laboratory of Pathology. In these lectures, either a differential approach by specific ultrastructural structures, or a global analysis of specific entities is offered. The residents are presented in advance with electron micrographs and/or light microscopic slides and are quizzed on certain structures or entities.
For more information on clinical departments (e.g., procedures, staff information, specimen collection, forms, etc), please see:
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Last Updated 1/12/2009 3:43:11 PM
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