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Diagnosis of B-cell non-hodgkin lymphomas with small-/intermediate-sized cells in cytopathology.

Schwock J, Geddie WR - Patholog Res Int (2012)

Bottom Line: Fine needle sampling is a fast, safe, and potentially cost-effective method of obtaining tissue for cytomorphologic assessment aimed at both initial triage and, in some cases, complete diagnosis of patients that present clinically with lymphadenopathy.Importantly, the recognition of specific cytologic features is crucial in guiding the appropriate selection of ancillary tests which will either confirm or refute a tentative diagnosis.We summarize the most pertinent cytomorphologic features for each entity as well as for reactive lymphoid hyperplasia, contrast them with each other to facilitate their recognition, and highlight common diagnostic pitfalls.

View Article: PubMed Central - PubMed

Affiliation: Division of Anatomical Pathology, Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto General Hospital, Room E11-219, Toronto, ON, Canada M5G 2C4.

ABSTRACT
Fine needle sampling is a fast, safe, and potentially cost-effective method of obtaining tissue for cytomorphologic assessment aimed at both initial triage and, in some cases, complete diagnosis of patients that present clinically with lymphadenopathy. The cytologic diagnosis of B-cell non-Hodgkin lymphomas composed of small-/intermediate-sized cells, however, has been seen as an area of great difficulty even for experienced observers due to the morphologic overlap between lymphoma and reactive lymphadenopathies as well as between the lymphoma entities themselves. Although ancillary testing has improved diagnostic accuracy, the results from these tests must be interpreted within the morphological and clinical context to avoid misinterpretation. Importantly, the recognition of specific cytologic features is crucial in guiding the appropriate selection of ancillary tests which will either confirm or refute a tentative diagnosis. For these reasons, we here review the cytologic characteristics particular to five common B-cell non-Hodgkin lymphomas which typically cause the most diagnostic confusion based on cytological assessment alone: marginal zone lymphoma, follicular lymphoma, mantle cell lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, and lymphoplasmacytic lymphoma. We summarize the most pertinent cytomorphologic features for each entity as well as for reactive lymphoid hyperplasia, contrast them with each other to facilitate their recognition, and highlight common diagnostic pitfalls.

No MeSH data available.


Related in: MedlinePlus

Hand position during the one-step smear with touch-off which allows multiple smears to be produced from a single fine-needle sample. The sample is expelled from the needle and placed near the frosted end of the “receiver” slide (1). The “spreader” slide (2) is then lowered in a hinge-like fashion onto the receiver slide from above, allowing the sample to disperse by capillary action, and then drawn down the length of the receiver slide producing the characteristic “bullet shape with feather edge.” If using the touch-off technique, the spreader slide is lowered to pick up a small amount of the specimen and then turned over so that a clean glass surface can be used to spread the remaining sample on the receiver slide. The picked up sample on the reverse of the spreader slide (arrow) is then used to generate a second smear.
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Related In: Results  -  Collection


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fig2: Hand position during the one-step smear with touch-off which allows multiple smears to be produced from a single fine-needle sample. The sample is expelled from the needle and placed near the frosted end of the “receiver” slide (1). The “spreader” slide (2) is then lowered in a hinge-like fashion onto the receiver slide from above, allowing the sample to disperse by capillary action, and then drawn down the length of the receiver slide producing the characteristic “bullet shape with feather edge.” If using the touch-off technique, the spreader slide is lowered to pick up a small amount of the specimen and then turned over so that a clean glass surface can be used to spread the remaining sample on the receiver slide. The picked up sample on the reverse of the spreader slide (arrow) is then used to generate a second smear.

Mentions: The obtained sample is then placed on a glass slide and a “splitting” or “touch-off technique” can be used to generate several matching smears from the same sample (Figure 2). After the sample is expelled onto the slide, the smear is produced by even and thin spreading of the cells with a second “spreader” slide using the so-called “one-step” technique [18]. During this step, it is important not to apply too much pressure which will destroy fragile cells. Ideally direct smears are subsequently processed using both Papanicolaou and Romanowsky-type stains on paired slides after alcohol fixation and air-drying, respectively. The two staining methods highlight distinct cellular features thereby producing complimentary diagnostic information. In general, alcohol fixation with Papanicolaou stain emphasizes nuclear detail whereas air-drying with Romanowsky-type stains demonstrates the cytoplasm more effectively. Also, Romanowsky-type stains differentially color DNA a metachromatic purple and RNA blue. Diff-Quik, a proprietary “quick” stain, or other nonproprietary modified staining methods are frequently used and are particularly suitable for immediate specimen assessment in situations where confirmation of specimen adequacy is crucial [19]. It should be noted, though, that the higher concentration of methylene blue present in Diff-Quik and similar rapid stains can sometimes cause fine blastic chromatin to appear darker and more mature than it otherwise would, and produce the appearance of “pseudonucleoli” in cells if staining is performed on an incompletely dried smear. May-Grünwald-Giemsa (MGG) stain provides more subtle coloration of nuclei and is superior for staining of cytoplasmic granulations. MGG is highly pH-dependent and may be difficult to standardize. Finally, cell suspensions are produced by aspiration of sterile physiological saline, phosphate-buffered saline solution, or cell culture medium from a tube through the sampling needle into the syringe. The cell suspension can then be used for ancillary testing purposes and either directly submitted for flow or slide-based cytometry or, after further processing as cytospin preparation or cell block, for immunocytochemistry or fluorescence in-situ hybridization (FISH). Cell suspensions are ideal material for more specialized molecular methods that may be required for diagnostic and research purposes including polymerase-chain reaction (PCR) and gene expression profiling. Very small amounts of residual cell suspension can be expelled onto an FTA card or paper for indefinite storage of DNA.


Diagnosis of B-cell non-hodgkin lymphomas with small-/intermediate-sized cells in cytopathology.

Schwock J, Geddie WR - Patholog Res Int (2012)

Hand position during the one-step smear with touch-off which allows multiple smears to be produced from a single fine-needle sample. The sample is expelled from the needle and placed near the frosted end of the “receiver” slide (1). The “spreader” slide (2) is then lowered in a hinge-like fashion onto the receiver slide from above, allowing the sample to disperse by capillary action, and then drawn down the length of the receiver slide producing the characteristic “bullet shape with feather edge.” If using the touch-off technique, the spreader slide is lowered to pick up a small amount of the specimen and then turned over so that a clean glass surface can be used to spread the remaining sample on the receiver slide. The picked up sample on the reverse of the spreader slide (arrow) is then used to generate a second smear.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3368210&req=5

fig2: Hand position during the one-step smear with touch-off which allows multiple smears to be produced from a single fine-needle sample. The sample is expelled from the needle and placed near the frosted end of the “receiver” slide (1). The “spreader” slide (2) is then lowered in a hinge-like fashion onto the receiver slide from above, allowing the sample to disperse by capillary action, and then drawn down the length of the receiver slide producing the characteristic “bullet shape with feather edge.” If using the touch-off technique, the spreader slide is lowered to pick up a small amount of the specimen and then turned over so that a clean glass surface can be used to spread the remaining sample on the receiver slide. The picked up sample on the reverse of the spreader slide (arrow) is then used to generate a second smear.
Mentions: The obtained sample is then placed on a glass slide and a “splitting” or “touch-off technique” can be used to generate several matching smears from the same sample (Figure 2). After the sample is expelled onto the slide, the smear is produced by even and thin spreading of the cells with a second “spreader” slide using the so-called “one-step” technique [18]. During this step, it is important not to apply too much pressure which will destroy fragile cells. Ideally direct smears are subsequently processed using both Papanicolaou and Romanowsky-type stains on paired slides after alcohol fixation and air-drying, respectively. The two staining methods highlight distinct cellular features thereby producing complimentary diagnostic information. In general, alcohol fixation with Papanicolaou stain emphasizes nuclear detail whereas air-drying with Romanowsky-type stains demonstrates the cytoplasm more effectively. Also, Romanowsky-type stains differentially color DNA a metachromatic purple and RNA blue. Diff-Quik, a proprietary “quick” stain, or other nonproprietary modified staining methods are frequently used and are particularly suitable for immediate specimen assessment in situations where confirmation of specimen adequacy is crucial [19]. It should be noted, though, that the higher concentration of methylene blue present in Diff-Quik and similar rapid stains can sometimes cause fine blastic chromatin to appear darker and more mature than it otherwise would, and produce the appearance of “pseudonucleoli” in cells if staining is performed on an incompletely dried smear. May-Grünwald-Giemsa (MGG) stain provides more subtle coloration of nuclei and is superior for staining of cytoplasmic granulations. MGG is highly pH-dependent and may be difficult to standardize. Finally, cell suspensions are produced by aspiration of sterile physiological saline, phosphate-buffered saline solution, or cell culture medium from a tube through the sampling needle into the syringe. The cell suspension can then be used for ancillary testing purposes and either directly submitted for flow or slide-based cytometry or, after further processing as cytospin preparation or cell block, for immunocytochemistry or fluorescence in-situ hybridization (FISH). Cell suspensions are ideal material for more specialized molecular methods that may be required for diagnostic and research purposes including polymerase-chain reaction (PCR) and gene expression profiling. Very small amounts of residual cell suspension can be expelled onto an FTA card or paper for indefinite storage of DNA.

Bottom Line: Fine needle sampling is a fast, safe, and potentially cost-effective method of obtaining tissue for cytomorphologic assessment aimed at both initial triage and, in some cases, complete diagnosis of patients that present clinically with lymphadenopathy.Importantly, the recognition of specific cytologic features is crucial in guiding the appropriate selection of ancillary tests which will either confirm or refute a tentative diagnosis.We summarize the most pertinent cytomorphologic features for each entity as well as for reactive lymphoid hyperplasia, contrast them with each other to facilitate their recognition, and highlight common diagnostic pitfalls.

View Article: PubMed Central - PubMed

Affiliation: Division of Anatomical Pathology, Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto General Hospital, Room E11-219, Toronto, ON, Canada M5G 2C4.

ABSTRACT
Fine needle sampling is a fast, safe, and potentially cost-effective method of obtaining tissue for cytomorphologic assessment aimed at both initial triage and, in some cases, complete diagnosis of patients that present clinically with lymphadenopathy. The cytologic diagnosis of B-cell non-Hodgkin lymphomas composed of small-/intermediate-sized cells, however, has been seen as an area of great difficulty even for experienced observers due to the morphologic overlap between lymphoma and reactive lymphadenopathies as well as between the lymphoma entities themselves. Although ancillary testing has improved diagnostic accuracy, the results from these tests must be interpreted within the morphological and clinical context to avoid misinterpretation. Importantly, the recognition of specific cytologic features is crucial in guiding the appropriate selection of ancillary tests which will either confirm or refute a tentative diagnosis. For these reasons, we here review the cytologic characteristics particular to five common B-cell non-Hodgkin lymphomas which typically cause the most diagnostic confusion based on cytological assessment alone: marginal zone lymphoma, follicular lymphoma, mantle cell lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma, and lymphoplasmacytic lymphoma. We summarize the most pertinent cytomorphologic features for each entity as well as for reactive lymphoid hyperplasia, contrast them with each other to facilitate their recognition, and highlight common diagnostic pitfalls.

No MeSH data available.


Related in: MedlinePlus