Protein Electrophoresis

LEARNING OBJECTIVES

After completing this training program, the user will be able to:

• Describe the methods of agarose gel electrophoresis, immunofixation, and densitometry.
• Name the clinical conditions most commonly associated with monoclonal gammopathy, and some key clinical features of each condition.
• View an agarose gel, an immunofixation, and a densitometry and determine if a monoclonal gammopathy is present. If it is present, the user will be able to identify the immunoglobulin type.
• View an agarose gel of serum, urine, or cerebrospinal fluid and identify the electrophoresis pattern (e.g., inflammation, liver disease, hemolysis, alpha-1 antitrypsin deficiency, tubular proteinuria, CSF oligoclonal banding, etc).

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  AUTHORS Michael L. Astion, MD, PhD Joseph P. Rank, MD Mark H. Wener, MD Lawrence M. Killingsworth, PhD

  CONTINUING EDUCATION Course # : 466-003-03 Contact Hours : 3

Urinalysis

LEARNING OBJECTIVES

• Describe the anatomic structures involved in urine formation.
• Describe the three parts of a complete urinalysis.
• Describe three methods of enhancing the visualization of urinary sediment structures.
• Identify and differentiate the common cell types found in urinary sediment.
• Enumerate red blood cells and white blood cells in unstained urinary sediment.
• Identify the types of casts seen in urinary sediment and state the clinical significance associated with each finding.
• Differentiate between crystals found in normal urine and crystals associated with clinical disease.
• Describe typical urinary sediment findings and key biochemical findings associated with selected renal disorders.

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  AUTHORS Carla M. Phillips, MLM, MT(ASCP) Paul J. Henderson, BS, MT(ASCP) Claudia Bien, MT(ASCP) James S. Fine, MD Adam R. Orkand, BA Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-004-03 Contact Hours : 2

Peripheral Blood

LEARNING OBJECTIVES

• Describe the steps necessary for the preparation of a Wright-stained peripheral blood smear and its proper microscopic evaluation.
• Identify the sequence of cell types encountered during normal maturation of myeloid, erythroid, and megakaryocytic cells in the marrow.
• Recognize and name the normal and abnormal forms of neutrophils, erythrocytes, lymphocytes, macrophages, and platelets using proper medical terminology.
• Correlate single morphologic abnormalities seen in any of the above cell lineages to one or more specific disease states.
• Integrate combinations of morphologic abnormalities involving one or more cell lineages to suggest a diagnosis of a disease state.

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  AUTHORS Brent L. Wood, MD, PhD Janet D. Curtis, BS, MT(ASCP) Joyce A. Behrens, MS, MT(ASCP) Adam R. Orkand, BA Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-002-03 Contact Hours : 2

Introduction to Transfusion Service

LEARNING OBJECTIVES

• Name the 4 components that can be made from a unit of whole blood and describe the function of each.
• Define the following terms: antigen, clinically significant antibody, hemolysis, and the complement system.
• List compatible components for the following patient blood groups: A, B, O, and AB.
• Define the antibody screen and explain why it is important in pre-transfusion testing.
• Name 3 crossmatch tests and define when an antiglobulin crossmatch must be done.
• Describe the laboratory's role in the evaluation of a suspected transfusion reaction.
• Define the elements of accurate and thorough testing prior to infusion of blood components to a patient.

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  AUTHORS Theresa Nester, MD Adam Orkand, BA EDITORS Michael Astion Dawn Rumsey Darolyn Crandall Jon Anderson Anne Zimmerman Joyce Behrens

  CONTINUING EDUCATION Course # : 466-023-03 Contact Hours : 2

Gram Stain

LEARNING OBJECTIVES

• State the clinical importance of the Gram stain of body fluids, and describe and state the purpose of each step in the procedure.
• List the major organisms isolated from the following specimens: blood, CSF, urine, respiratory, genital, wound, joint, eye, and stool.
• State whether a specimen is likely to be normal, contaminated, or infected.
• Use the proper generic terminology to systematically describe the morphology of bacteria or yeast.
• Use the proper terminology to identify and describe cell types and common artifacts.

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  AUTHORS Brad T. Cookson, MD, PhD Ajit Limaye, MD Janet Curtis, BS, MT(ASCP) Thomas R. Fritsche, MD, PhD Lee Anne McGonagle, MPH, SM(AAM) Adam R. Orkand, BA Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-008-03 Contact Hours : 2

Mycology

LEARNING OBJECTIVES

• Recognize the four classes of fungi and the sexual cycles used to divide each group.
• Demonstrate the ability to use color, shape, texture, and differential media and tests to accurately and correctly describe a specimen.
• Describe in detail the most common procedures to identify yeasts and molds.
• Recognize further tests that are available if necessary.
• List the conidiogenesis methods commonly used in the laboratory to recognize and define different molds.
• List the common superficial, subcutaneous, and systemic mycoses and recognize the clinical presentation, obverse and reverse colonies, and microscopic features of each.
• Identify macroscopically and microscopically common opportunistic fungi as well as some of the less common opportunists.

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  AUTHORS Marie B. Coyle, PhD Dave Nowowiejski, MS, MT(ASCP) Karen LaFe, MT(ASCP) Deborah Ritter, MS, CLS(NCA) Janet D. Curtis, BS, MT(ASCP) Adam R. Orkand, BA Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-005-03 Contact Hours : 2

Parasitology

LEARNING OBJECTIVES

• Explain common parasite life cycles and relate how that knowledge aids in selection of the appropriate test methodology.
• Discuss the different lab tests available for detection of parasites and compare each in detail.
• Discuss how a particular laboratory approach is chosen to detect a certain parasite and why it is the best choice.
• Demonstrate the ability to interpret the results of the various methods of identifying the presence of parasites, both intracellular and extracellular in blood and in stool samples.
• Discuss the general geographic distribution of many common parasites seen in the laboratory.
• Differentiate between true parasites and common artifacts that are seen and recognize when to pay special attention to the possibility of confusing the two.
• Identify parasites found in sites other than blood and stool samples and the appropriate way to test for each of them.

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  AUTHORS Thomas R. Fritsche, MD, PhD Sam C. Eng, MS, SM(AAM) Janet Curtis, BS, MT(ASCP) Adam R. Orkand, BA Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-006-03 Contact Hours : 3

Microscopy

LEARNING OBJECTIVES

• Explain the function of each part of the light microscope and show where it is located.
• Define "resolution", "contrast", and "definition" as they relate to the light microscope. Explain which parts of the microscope affect each. Manipulate these parts.
• The user will be able to summarize the optical path of light throughout the microscope from beginning to end.
• The user can determine the magnification, numerical aperture, and optimum coverslip thickness of an objective lens.
• The user can perform each step in the process of Köhler illumination in order. They can also use the fine focus control to visualize the third dimension of a specimen.
• The user can recognize when an objective lens is dirty and can perform this simple maintenance procedure.
• The user can understand common lens aberrations as evidenced by choosing the best optics for a given application. The user will also be capable of critically evaluating manufacturer's specifications.
• The user will have a spatially accurate, 3-D understanding of the three properties of light and how they relate to the light microscope.

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  AUTHORS Len Pagliaro, MD Adam R. Orkand, BA Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-11-03 Contact Hours : 2

Amniotic Fluid

LEARNING OBJECTIVES

• Describe the amniocentesis procedure and the most common indications for amniocentesis.
• List the most common genetic abnormalities detected by cytogenetic analysis and their clinical features.
• Describe the L/S ratio and other assays used to assess fetal lung maturity.
• Describe the tests used to screen for open neural tube defects.
• Describe hemolytic disease of the newborn (HDN) and how the severity of HDN is assessed in the lab.

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  AUTHORS Jessica D. Eisner, MD James S. Fine, MD Adam R. Orkand, BA Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-019-02 Contact Hours : 1

Cerebrospinal Fluid

LEARNING OBJECTIVES

• Describe the production and collection of cerebrospinal fluid (CSF).
• Describe the gross appearance of CSF specimens.
• Describe the principles of manual chamber counts, cytocentrifuge preparation, and Wright staining.
• Identify the major types of normal and abnormal cells in CSF and describe their signficance.

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  AUTHORS Brent Wood, MD, PhD Jana Roney, BS, MT(ASCP) Adam R. Orkand, BA Jessica Eisner, MD Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-018-02 Contact Hours : 1

Seminal Fluid

LEARNING OBJECTIVES

• List the glandular contributions and cellular components of seminal fluid.
• Describe the methods used to determine sperm concentration, motility and morphology.
• List the abnormalities detected and reference ranges for semen analysis.
• Discuss the goals and limitations of semen analysis in the clinical setting.

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  AUTHORS Brent Wood, MD, PhD Jana Roney, BS, MT(ASCP) Charles H. Muller, PhD Adam R. Orkand, BA Jessica Eisner, MD Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-013-03 Contact Hours : 2

Serous Fluid

LEARNING OBJECTIVES

• Describe the normal production of serous fluids and the mechanisms of formation of transudates and exudates.
• Distinguish transudates and exudates using lab testing, and understand their significance.
• Describe the gross appearance of serous fluid specimens.
• Describe the principles of manual chamber counts, cytocentrifuge preparation, and Wright staining.
• Identify the major types of normal and abnormal cells in serous fluids and their significance.

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  AUTHORS Brent Wood, MD, PhD Jana Roney, BS, MT(ASCP) Adam R. Orkand, BA Jessica Eisner, MD Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-020-02 Contact Hours : 1

Synovial Fluid

LEARNING OBJECTIVES

• Describe the basic classification of synovial effusions.
• Describe the gross appearance of synovial fluid specimens.
• Discuss the principles and applications of polarizing microscopy and compensated polarizing microscopy, and how to perform them.
• Distinguish urate crystals from calcium pyrophosphate dihydrate (CPPD) crystals.
• Describe other crystals, artifacts, inclusions, and cell types that may be present in synovial fluids.

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  AUTHORS Mark H. Wener, MD Adam R. Orkand, BA Jessica Eisner, MD Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-010-03 Contact Hours : 1

Immunology

Anti-nuclear Antibody (ANA)

LEARNING OBJECTIVES

• Identify the steps of an indirect immunofluorescence antibody (IFA) procedure and state the purpose of each step.
• Identify the steps of an enzyme immunoassay (EIA) procedure and state the purpose of each step.
• Describe the function of various components of an epifluorescence microscope and how they influence IFA staining.
• State the role of ANA testing in the diagnosis of autoimmune rheumatic diseases.
• View HEp-2 IFA results and determine whether they should be called ANA positive or negative in comparison to an endpoint control.
• Identify the correct ANA titer when presented with a series of images.
• Identify interphase (resting) and mitotic (dividing) HEp-2 cells.
• Identify ANA staining patterns on HEp-2 cells:
  • Beginner: identify the common ANA patterns (homogeneous, speckled, nucleolar, centromere) by their specific name, and be able to differentiate them from uncommon patterns.
  • Advanced: identify common and uncommon ANA patterns by their specific name.
• Identify mixed ANA patterns:
  • Beginner: those that have one or more common ANA patterns.
  • Advanced: also identify those that have uncommon patterns.
  • All: state two strategies that can help identify mixed patterns.
• Identify cytoplasmic staining patterns on HEp-2 substrate:
  • Beginner: identify the staining as cytoplasmic.
  • Advanced: identify patterns that are likely to be mitochondrial, ribosomal, or Golgi.
• Identify artifacts on HEp-2.
• Identify positive results of IFA testing on Crithidia luciliae; state the antibody being tested, and the clinical utility of a positive result.
• List the specific autoantibodies that are commonly associated with a positive ANA.
  • Beginner: identify the antibodies by name and the ANA patterns they are associated with.
  • Advanced: also identify the disease associations.
• State the meaning of the term ENA and identify the antigens that are considered to be "ENAs".

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  AUTHORS Kathleen W. Hutchinson, MS, MT(ASCP) Mark H. Wener, MD Bruce G. Gilliland, MD Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-035-04 Contact Hours : 2

Mouse Stomach-Kidney (MSK)

LEARNING OBJECTIVES

• Identify the steps of an indirect immunofluorescence antibody (IFA) procedure and be able to state the purpose of each step.
• State the role of cytoplasmic antibody testing in the diagnosis of autoimmune liver disease and pernicious anemia.
• Identify the stomach mucosal tissue, the smooth muscle layer, the kidney cortex and medulla tissue.
• Identify parietal cells, chief cells, smooth muscle layer, renal tubules, and blood vessels on mouse stomach-kidney (MSK) tissue.
• View MSK IFA results and identify staining patterns.
  • Beginner: identify common staining patterns (mitochondrial, parietal cell, and smooth muscle) by their specific name, and differentiate them from the uncommon patterns.
  • Advanced: also identify uncommon patterns by their specific names.
• Identify clinical correlations, if any, that are associated with cytoplasmic antibodies. -Beginner: common staining patterns. -Advanced: both common and uncommon staining patterns.

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  AUTHORS Kathleen W. Hutchinson, MS, MT(ASCP) Mark H. Wener, MD Bruce G. Gilliland, MD Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-036-04 Contact Hours : 1

Anti-neutrophil Cytoplasmic Antibodies (ANCA)

LEARNING OBJECTIVES

• Identify the steps of an indirect immunofluorescence antibody (IFA) procedure and be able to state the purpose of each step.
• Identify the steps of an enzyme immunoassay (EIA) procedure and be able to state the purpose of each step.
• State the role of ANCA testing in the diagnosis of vasculitis.
• Identify neutrophils and lymphocytes on IFA substrates.
• View ANCA IFA results and identify C-ANCA, P-ANCA, ANA, mixed ANCA and ANA, and atypical staining patterns on ethanol fixed substrate.
• View ANCA IFA results and identify ANCA staining on formalin fixed substrate.
• State two strategies that can be used to differentiate P-ANCA, ANA staining, and mixed patterns.
• Identify the antibodies that are commonly associated with the C-ANCA and P-ANCA patterns.
• Identify the clinical correlations, if any, that are associated with ANCA staining patterns.

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  AUTHORS Kathleen W. Hutchinson, MS, MT(ASCP) Mark H. Wener, MD Bruce G. Gilliland, MD Michael L. Astion, MD, PhD

  CONTINUING EDUCATION Course # : 466-037-04 Contact Hours : 2