Protocol for the Examination of Specimens From Patients With Non-Hodgkin Lymphoma/Lymphoid Neoplasms

Imprint cytology: Invaluable technique to evaluate fresh specimens received in the pathology department for lymphoma workup

“…19,20 In cases suspicious for classic Hodgkin lymphoma, with an imprint showing cells highly suggestive of Hodgkin or Reed-Sternberg cells, formalin fixation and paraffin-embedding (FFPE) should be prioritized if the specimen is very limited because well-fixed sections and immunophenotyping by immunohistochemistry are necessary in the diagnosis of nearly all cases of Hodgkin lymphoma, whereas FCM, molecular, and FISH studies are less likely to contribute. 9 If sufficient tissue is available, small fragments of fresh lymphoid specimens can be snap frozen and stored at -80°C for molecular analyses of B-or T-cell clonality or for some of the translocations, such as t(14;18) and t (11;14) in follicular lymphoma and mantle cell lymphoma, respectively, although FISH remains more sensitive than PCR for these rearrangements. 8,21,22 Prioritization at triage is crucial: In all situations, including with very small specimens, the priority remains formalin fixation to obtain H&E-stained sections for diagnosis and ancillary studies, most of which can be performed from the FFPE tissue block, including immunohistochemistry, FISH, and molecular studies.…”

“…9 If sufficient tissue is available, small fragments of fresh lymphoid specimens can be snap frozen and stored at -80°C for molecular analyses of B-or T-cell clonality or for some of the translocations, such as t(14;18) and t (11;14) in follicular lymphoma and mantle cell lymphoma, respectively, although FISH remains more sensitive than PCR for these rearrangements. 8,21,22 Prioritization at triage is crucial: In all situations, including with very small specimens, the priority remains formalin fixation to obtain H&E-stained sections for diagnosis and ancillary studies, most of which can be performed from the FFPE tissue block, including immunohistochemistry, FISH, and molecular studies. Thus, once the specimen has been triaged as described above, and to maximize tissue for ancillary studies, sections from the excision specimen or the cores are divided into several blocks, ideally 1 thin section or 1 core per block, and fixed in formalin.…”

“…When there is tissue for more than 1 to 2 blocks, we put part in zinc-sulfate fixative because it yields superior cytomorphologic details and remains amenable for immunohistochemistry. 8,16…”

“…In such cases, or when there is clinical suspicion for classic Hodgkin lymphoma, FFPE tissue should be prioritized because histology and immunohistochemistry are essential for diagnosis. 9 A monomorphous population of lymphoid cells raises the possibility of a lymphoproliferative disorder; as in most non-Hodgkin lymphomas, particularly of B-cell phenotype, the range of lymphoid cellular sizes is more limited. 10,23,30 Such infiltrates on imprints may be broadly subdivided into lesions composed mostly of small-, intermediate-, or large-size lymphocytes.…”

“…Many institutions have their own established protocols; in addition, the College of American Pathologists has issued recommendations and a protocol for the examination of specimens from patients with lymphoma. 8,9 Past publications describe the use of cytologic preparations (smears and imprints) for intraoperative assessment of lymph nodes or lymphoid lesions to facilitate the identification of entities not as easily evaluable in conventional frozen sections: The clarity of the cytologic details enables the identification of Hodgkin lymphomas, non-Hodgkin lymphomas, and certain metastatic tumors. [10][11][12][13] In contrast to smears, imprints are easily prepared by gently touching the fresh-cut surface of tissues to a glass slide, without smearing, with minimal distortion and without damaging fragile core biopsies.…”