Practical Flow Cytometry in Haematology

100 Worked Examples

Mike Leach, Mark Drummond, Allyson Doig, Pam McKay, Bob Jackson, Barbara J. Bain

• Publisher: Wiley-Blackwell
• Year: 2015
• ISBN:
  9781118746899
• Edition: 1
• Topic: Medicina
• Subtopic: Hematología

The analysis of blood, bone marrow and tissue fluid specimens requires a multi-faceted approach with the integration of scientific data from a number of disciplines. No single discipline can operate in isolation or errors will occur. Flow cytometry is in a privileged position in that it can provide rapid analysis of specimens and it is often the first definitive investigation to produce results and help formulate a working diagnosis.

This companion text to Practical Flow Cytometry in Haematology Diagnosis contains 100 worked examples drawn from real clinical cases presenting to the authors' institution. Cases are illustrated with peripheral blood and bone marrow cytology, tissue pathology and cytogenetic and molecular data, which are integrated to generate, where appropriate, a diagnosis based on the WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. The spectrum of clinical cases includes adult and paediatric patients, and both neoplastic and reactive disorders. The cases appear in no particular order to challenge the reader to make their own diagnosis.

The reader will review May?Grünwald?Giemsa (MGG)-stained films of peripheral blood and bone marrow aspirates presented alongside flow cytometric data and haematoxylin and eosin (H&E)-stained bone marrow and other tissue biopsy sections. Immunohistochemistry is used to further clarify the tissue lineage and cell differentiation. Cytogenetic studies using metaphase preparations are used to identify translocations and chromosome gains and losses whilst interphase fluorescence in situ hybridisation (FISH) studies and polymerase chain reaction (PCR) are used to identify gene fusions, gene rearrangements and deletions. Each case concludes with a discussion of the features that are important to making a diagnosis. The cases are also listed according to disease classification in the appendix so that the text can also be used as a reference.

Practical Flow Cytometry in Haematology: 100 Worked Examples:

• Provides a practical, example-based resource for flow cytometry
• Demonstrates how flow cytometry results should be interpreted and applied to optimize patient care
• Includes both malignant and benign conditions
• Can be used in conjunction with Practical Flow Cytometry in Haematology Diagnosis, by the same author team (ISBN 9780470671207)

Practical Flow Cytometry in Haematology: 100 Worked Examples is ideal for practicing haematologists and histopathologists with an interest in haematopathology, but particularly directed at trainee haematologists and scientists preparing for FRCPath and related examinations.

Case 1

An 11-year-old boy was admitted with a short history of fever, sweats, dyspnoea and left chest discomfort. There was no past history of note. Examination identified features of a left pleural effusion. There was also a tender swelling of the left anterior chest in the upper pectoral region and palpable cervical lymphadenopathy. The liver and spleen were not palpable.

Laboratory investigations

FBC and blood film: normal

U&Es, LFTs: normal. LDH was 1460 U/L.

Imaging

The CXR showed opacification and loss of aeration of the left hemithorax in keeping with a pleural effusion (Figure 1.1). CT imaging confirmed this but in addition identified a left pleural-based mass, abnormal soft tissue in the left pectoral muscles (arrows, Figure 1.2) and cervical lymphadenopathy. In addition, there was collapse/consolidation of the lower left lung, creating the appearance of an air bronchogram. A core biopsy of a cervical node was taken and the pleural effusion was aspirated for analysis.

Figure 1.1 CXR.

Figure 1.2 CT.

Flow cytometry

The pleural fluid cell count was 0.98 × 10⁹/L. A cytospin preparation showed three distinct cell types: a small mature lymphoid population in keeping with reactive lymphocytes, an intermediate sized/large sized lymphoid population and a large cell population with pleomorphic morphology and blue cytoplasm (Figures 1.3–1.6). The cells with the abundant cytoplasm (Figures 1.3 and 1.4) and the single binucleate cell (Figure 1.6) are reactive mesothelial cells. The cells with the cytoplasmic blebs (Figures 1.4–1.6) are the disease cells, which were the subsequent focus for immunophenotyping studies.

Figure 1.3 MGG, ×500.

Figure 1.4 MGG, ×500.

Figure 1.5 MGG, ×500.

Figure 1.6 MGG, ×500.

By applying a blast gate to the suspected malignant cells in the FSC/SSC analysis (Figure 1.7), they were shown to express CD45^bright (Figure 1.8), CD2 (Figure 1.9), cCD3 [whilst surface CD3 was negative apart from a few reactive T cells (Figure 1.8)], partial CD7 (Figure 1.10) and CD13. Othe...