Flow Cytometry Laboratory

The Clinical Flow Cytometry Laboratory at Massachusetts General Hospital provides pathologists and clinicians with results that directly impact the care of patients with hematologic disorders as well as patients with other  disorders that require understanding of an individual’s immune system. 

The lab utilizes BD Biosciences Flow Cytometers, capable of analyzing up to 10-color immunofluorescent data and cellular light scatter. Specimen preparation instruments include two BD Lyse/Wash Assistants and a BD Sample Prep Assistant. This CLIA certified laboratory is currently the workplace of 3 full time medical technologists as well as a laboratory supervisor and occupies a workspace located in the 5th floor of the Warren Building, Room 506.

The Director of the lab, Dr. Frederic Preffer PhD, may be located in his office on the 4th floor of the Simches building, Room 4226 when he is not in one of his research or clinical laboratories or reviewing cases with pathologists and residents.

Faculty

Frederic Preffer, MS, PhD, Director, Flow Cytometry Lab

The laboratory serves the entire Mass General clinical community, as well as outside cases from MEEI, The North Shore Cancer Center and other institutions. The majority of specimens include those obtained from peripheral blood, lymph nodes and bone marrow. However, fluids from various body cavities, [e.g., spinal, thoracic fluids] account for a significant number of additional cases. The laboratory works closely with the department's cytopathology service in immunophenotyping FNA's of lymph nodes and other hematopoietic tissue as well as lymph node and other tissue biopsy specimens obtained in the frozen section laboratory.

From the laboratory’s inception in 1996 through current years, the number of cases has increased significantly. Much of the laboratory growth has been appreciated in the form of additional new tests offered. In 2020 the laboratory processed the following types of cases and associated case counts: Leukemia/Lymphoma (3845), T-cell subsets (2301), Immunodeficiency (2670), Peripheral blood stem cell/apheresis collections (732), Organ Transplant T-cell monitoring (97) and “other” miscellaneous tests (650). In order to accommodate the prepping/staining/acquisition/analysis of data from such a heavy sample workload with only three technologists, the laboratory continues to implement strategies to automate processes, batch runs and utilize techniques to reduce time and mistakes from slowing down sample processing. The laboratory prides itself on quick turnaround time and the ability to provide STAT turn-around times to clinicians who are treating patients who are in acute clinical situations.

Academic and Research Accomplishments

Although the clinical Flow Cytometry Laboratory is primarily a service laboratory, it has made significant contributions to the body of published clinical literature, utilizing flow cytometry in numerous useful ways.

  1. Graeme-Cook F, Bell DA, Flotte TJ, Preffer FI, Pastel-Levy C, Nardi G, Compton C. Aneuploidy in pancreatic insulinomas does not predict malignancy. Cancer 1990; 66:2365-2368. doi: 10.1002/1097-0142(19901201)66:11<2365::aid-cncr2820661119>3.0.co;2-1.PMID: 2245392
  2. Pastel-Levy C, Flotte TJ, Preffer F, Ware A, Graeme-Cook F, Bell DA, Colvin RB. Application of DNA flow cytometry from paraffin-embedded tissue to the diagnosis of mycosis fungoides. J Cutan Pathol 1991; 18:279-283. doi: 10.1111/j.1600-0560.1991.tb01236.x.PMID: 1939787
  3. Colvin RB, Preffer FI. Laboratory monitoring of therapy with OKT3 and other murine monoclonal antibodies. Clin Lab Med 1991; 11:693-714.PMID: 1934971
  4. Eichhorn JH, Bell DA, Young RH, Swymer CM, Flotte TJ, Preffer FI, Scully RE. DNA content and proliferative activity in ovarian small cell carcinomas of the hypercalcemic type: Implications for diagnosis, prognosis and histogenesis. Am J Clin Pathol 1992; 98:579-586. doi: 10.1093/ajcp/98.6.579.PMID: 1334364
  5. Flotte TJ, Pastel-Levy C, Graeme-Cook F, Preffer FI, Bell DA. DNA flow cytometry of primary tumors and their skin metastases. Correlation of DNA histograms. Am J Dermatopathol 1992; 14:19-23. doi: 10.1097/00000372-199202000-00003.PMID: 1510211
  6. Hug EB, Donnelly SM, Shipley WU, Heney NM, Kaufman DS, Preffer FI, Schwartz SM, Colvin RB, Althausen AF. Deoxyribonucleic acid flow cytometry in invasive bladder carcinoma: a possible predictor for successful bladder preservation following transurethral surgery and chemotherapy-radiotherapy. J Urol 1992; 148:47-51. doi: 10.1016/s0022-5347(17)36505-9.PMID: 1613879
  7. Jacoby AF, Young RH, Colvin RB, Flotte TJ, Preffer FI, Scully RE, Swymer CM, Bell DA. DNA content in juvenile granulosa cell tumors of the ovary: a study of early- and advanced-stage disease. Gynecol Oncol 1992; 46:97-103. doi: 10.1016/0090-8258(92)90203-u.PMID: 1634146
  8. Kurban RS, Preffer FI, Sober AJ, Mihm MC, Barnhill RL. Occurrence of melanoma in “dysplastic” nevus spilus: report of case and analysis by flow cytometry. J Cutan Pathol 1992; 19:423-428. doi: 10.1111/j.1600-0560.1992.tb00615.x.PMID: 1474193
  9. Pastel-Levy C, Bell DA, Rosenberg AE, Preffer FI, Colvin RB, Flotte TJ. DNA flow cytometry of epithelioid sarcoma. Cancer 1992; 70:2823-2826. doi: 10.1002/1097-0142(19921215)70:12<2823::aid-cncr2820701216>3.0.co;2-9.PMID: 1451061
  10. Centeno BA, Louis DN, Kupsky WJ, Preffer FI, Sobel RA. The AgNOR technique, PCNA immunohistochemistry, and DNA ploidy in the evaluation of choroid plexus biopsy specimens. Am J Clin Pathol 1993; 100:690-696. doi: 10.1093/ajcp/100.6.690.PMID: 7902669
  11. Oyama T, Vickery AL, Preffer FI, Colvin RB. A comparative study of flow cytometry and histopathologic findings in thyroid follicular carcinomas and adenomas. Hum Pathol 1994; 25:271-275. doi: 10.1016/0046-8177(94)90199-6.PMID: 8150457
  12. Centeno BA, Zietman AL, Shipley WU, Sobczak ML, Shipley JW, Preffer FI, Boyle BJ, Colvin RB. Flow cytometric analysis of DNA ploidy, percent S-phase fraction, and total proliferative fraction as prognostic indicators of local control and survival following radiation therapy for prostate carcinoma. Int J Radiat Oncol Biol Phys 1994; 30:309-315. doi: 10.1016/0360-3016(94)90009-4.PMID: 7928459
  13. Quintanilla-Martinez L, Preffer F, Rubin D, Ferry JA, Harris NL. CD20+ T-cell lymphoma. Neoplastic transformation of a normal T-cell subset. Am J Clin Pathol 1994; 102:483-489. doi: 10.1093/ajcp/102.4.483.PMID: 7524302
  14. Chae YS, Flotte T, Hsu DW, Preffer F, Hedley-Whyte ET. Flow cytometric DNA ploidy and cells phase fractions in recurrent human pituitary adenomas. A correlative study of flow cytometric analysis and the expression of proliferating cell nuclear antigen. Gen Diagn Pathol 1996; 142:89-95.PMID: 8950573
  15. Cheek RF, Olszak I, Madoff S, Preffer FI. In vitro detection of Mycoplasma fermentans binding to B-lymphocytes in fresh peripheral blood using flow cytometry. Cytometry 1997; 28:90-95. doi: 10.1002/(sici)1097-0320(19970501)28:1<90::aid-cyto11>3.0.co;2-m.PMID: 9136760
  16. Nguyen PL, Olszak I, Harris NL, Preffer FI. Myeloperoxidase detection by three-color flow cytometry and by enzyme cytochemistry in the classification of acute leukemia. Am J Clin Pathol 1998; 110:163-169. doi: 10.1093/ajcp/110.2.163.PMID: 9704614
  17. Catlin EA, Roberts JD, Erana R, Preffer FI, Ferry JA, Kelliher AS, Atkins L, Weinstein HJ. Transplacental transmission of natural-killer-cell lymphoma. N Eng J Med 1999; 341:85-91. doi: 10.1056/NEJM199907083410204.PMID: 10395632
  18. Treon SP, Shima Y, Preffer FI, Doss DS, Ellman L, Schlossman RL, Grossbard ML, Belch AR, Pilarski LM, Anderson KC. Treatment of plasma cell dyscrasias by antibody-mediated immunotherapy. Semin Oncol 1999; 26:97-106.PMID: 10561024
  19. Treon SP, Shima Y, Grossbard ML, Preffer FI, Belch AR, Pilarski LM, Anderson KC. Treatment of multiple myeloma by antibody mediated immunotherapy and induction of myeloma selective antigens. Ann Oncol 2000; 11:107-111.PMID: 10707790
  20. Dong HY, Harris NL, Preffer FI, Pitman MB. Fine-needle aspiration biopsy in the diagnosis and classification of primary and recurrent lymphoma: a retrospective analysis of the utility of cytomorphology and flow cytometry. Mod Pathol 2001; 14:472-481. doi: 10.1038/modpathol.3880336.PMID: 11353059
  21. Treon SP, Pilarski LM, Belch AR, Kelliher A, Preffer FI, Shima Y, Mitsiades CS, Mitsiades NS, Szczepek AJ, Ellman L, Harmon D, Grossbard ML, Anderson KC. CD20-directed serotherapy in patients with multiple myeloma: biologic considerations and therapeutic applications. J Immunother 2002; 25:72-81. doi: 10.1097/00002371-200201000-00008.PMID: 11924912
  22. Treon SP, Kelliher A, Keele B, Frankel S, Emmanouilides C, Kimby E, Schlossman R, Mitsiades N, Mitsiades C, Preffer F, Anderson KC. Expression of serotherapy target antigens in Waldenstrom’s macroglobulinemia: therapeutic applications and considerations. Semin Oncol 2003; 30:248-252. doi: 10.1053/sonc.2003.50047.PMID: 12720146
  23. Olson DP, Dombkowski DM, Kelliher AS, Pontillo C, Anderson DC, Preffer FI. Differential expression of cell surface antigens on subsets of CD4+ and CD8+ T cells. Med Sci Monit 2004; 10:BR339-BR345. Epub 2004 Aug 20.PMID: 15328479
  24. Treon SP, Emmanouilides C, Kimby E, Kelliher A, Preffer F, Branagan AR, Anderson KC, Frankel SR; Waldenstrom’s Macroglobulinemia Clinical Trials Group. Extended rituximab therapy in Waldenstrom’s macroglobulinemia. Ann Oncol 2005; 16:132-138. doi: 10.1093/annonc/mdi022.PMID: 15598950 Clinical Trial
  25. Kelliher AS, Parent DW, Anderson DC, Dorn ME, Hahn JL, Eapen S, Preffer FI. Novel use of the BD FACS SPA to automate custom monoclonal antibody panel preparations for immunophenotyping. Cytometry B Clin Cytom 2005; 66:40-45. doi: 10.1002/cyto.b.20055.PMID: 15918157
  26. Rahemtullah A, Reichard KK, Preffer FI, Harris NL, Hasserjian RP. A double-positive CD4+CD8+ T-cell population is commonly found in nodular lymphocyte predominant Hodgkin lymphoma. Am J Clin Pathol 2006; 126:805-814. doi: 10.1309/Y8KD-32QG-RYFN-1XQX.PMID: 17050078
  27. Ferry JA, Fung CY, Zukerberg L, Lucarelli MJ, Hasserjian RP, Preffer FI, Harris NL. Lymphoma of the ocular adnexa: A study of 353 cases. Am J Surg Pathol 2007; 31:170-184. doi: 10.1097/01.pas.0000213350.49767.46.PMID: 17255761
  1. Rahemtullah A, Harris N, Dorn ME, Preffer F, Hasserjian RP. Beyond the L&H cell: CD4+CD8+ T cells in nodular lymphocyte predominant Hodgkins lymphoma. Leuk Lymphoma 2008; 49:1870-1878. doi: 10.1080/10428190802308728.PMID: 18949610
  2. Rahemtullah A, Longtine JA, Harris NL, Dorn M, Zembowicz A, Quintanilla-Fend L, Preffer FI, Ferry JA. CD20+ T-cell lymphoma: clinicopathologic analysis of 9 cases and a review of the literature. Am J Surg Pathol 2008; 32:1593-1607. doi: 10.1097/PAS.0b013e31817d7452.PMID: 18753947
  3. Cserti-Gazdewich C, Dzik W, Dorn M, Quagliaroli R, Xu S, Ssewanyana I, Nayyar R, Preffer F. Quantitation of CD36 (platelet glycoprotein IV) expression on platelets and monocytes by flow cytometry: application to the study of Plasmodium falciparum malaria. Cytometry B Clin Cytom 2009; 76B:127-134. doi: 10.1002/cyto.b.20443.PMID: 18671254
  4. Cannizzo E, Sohani AR, Ferry JA, Hochberg EP, Kluk MJ, Dorn ME, Sadowski C, Bucci JJ, Ackerman AM, Longtine JA, Carulli G, Preffer FI. Carcinoma and multiple lymphomas in one patient: establishing the diagnoses and analyzing risk factors. J Hematop 2009; 23:163-170. doi: 10.1007/s12308-009-0041-0.PMID: 20309424
  5. Dzik WH, Cserti-Gazdewich C, Ssewanyana I, DeLelys M, Preffer FI. When monocytes and platelets compete: the effect of platelet count on the flow cytometric measurement of monocyte CD36. Cytometry B Clin Cytom 2010; 78:81-87. doi: 10.1002/cyto.b.20504.PMID: 19902557
  6. Cannizzo E, Bellio E, Sohani AR, Hasserjian RP, Ferry JA, Dorn ME, Sadowski C, Bucci JJ, Carulli G, Preffer FI. Multiparameter immunophenotyping by flow cytometry in multiple myeloma: the diagnostic utility of defining ranges of normal antigenic expression in comparison to histology. Cytometry B Clin Cytom 2010; 78B:231-238. doi: 10.1002/cyto.b.20517.PMID: 20198608
  7. Burdo TH, Lo J, Abbara S, Wei J, DeLelys M, Preffer F, Rosenberg ES, Williams KC, Grinspoon S. Soluble CD163, a novel marker of activated macrophages, is elevated and associated with noncalcified coronary plaque in HIV-infected patients. J Infect Dis 2011; 204:1227-1236. doi: 10.1093/infdis/jir520.PMID: 21917896
  8. Cannizzo E, Carulli G, Del Vecchio L, Ottaviano V, Bellio E, Zenari E, Azzara A, Petrini M, Preffer F. The role of CD19 and CD27 in the diagnosis of multiple myeloma by flow cytometry: a new statistical model. Am J Clin. Pathol. 2012; 137:377-386. doi: 10.1309/AJCP63TOCFNAMDMS.PMID: 22338049
  9. DeLelys ME, Ochoa G, Cserti-Gazdewich CM, Vietz C, Preffer FI and Dzik W. Relationship between ABO genotype and A antigen expression on platelets. Transfusion 2013; 53:1763-71. doi: 10.1111/j.1537-2995.2012.03952.x. Epub 2012 Nov 12.PMID: 23145947
  10. Platt MY, DeLelys ME, Preffer FI, Sohani AR. Flow cytometry is of limited utility in the early identification of ‘Double-Hit’ B-cell lymphomas. Cytometry B Clin Cytom 2013; 84:143-8. doi: 10.1002/cyto.b.21076. Epub 2013 Jan 22.PMID: 23341189
  11. Fathi AT, Preffer FI, Sadrzadeh H, Ballen KK, Amrein PC , Attar EC , McAfee SL,
  12. Kovach AE, DeLelys ME, Kelliher AS, Dillon LJ, Hasserjian RP, Ferry JA, Preffer FI, Sohani AR. Diagnostic utility of cerebrospinal fluid flow cytometry in patients with and without prior hematologic malignancy. Am J Hematol 2014; 89:978-84. doi: 10.1002/ajh.23806. Epub 2014 Jul 31.PMID: 25042070
  13. Ferry JA, Klepeis V, Sohani AR, Harris NL, Preffer FI, Stone JH, Grove A, Deshpande V. IgG4-related orbital disease and its mimics in a Western population. Am J Surg Pathol. 2015; 39:1688-1700. doi: 10.1097/PAS.0000000000000497.PMID: 26379149
  14. Aisagbonhi O, DeLelys M, Hartford N, Preffer F, Ly A. Utility of flow cytometry in diagnosing hematologic malignancy in tonsillar tissue.  Int J Surg Pathol 2017; 25(5):406-413. doi: 10.1177/1066896917700726. Epub 2017 Mar 28.PMID: 28351195
  15. Scarfo I, Irmhoj M, Frigault MJ, Castano AP, Lorrey S, Bouffard AA, van Scoyk A, Rodig SJ, Shay AJ, Aster JC, Preffer FI, Weinstock DM, Maus MV. Anti-CD37 chimeric antigen receptor T cells are active against B and T cell lymphomas.  Blood 2018; 132(14):1495-1506. doi: 10.1182/blood-2018-04-842708. Epub 2018 Aug 8.PMID: 30089630
  1. Narayan R, Blonquist TM, Emadi A, Hasserjian RP, Burke M, Lescinskas C, Neuberg DS, Brunner AM, Hobbs G, Hock H, McAfee SL, Chen YB, Attar E, Graubert TA, Bertoli C, Moran JA, Bergeron MK, Foster JE, Ramos AY, Som TT, Vartanian MK, Story JL, McGregor K, Macrae M, Behnan T, Wey MC, Rae J, Preffer FI, Lesho P, Duong VH, Mann ML, Ballen KK, Connolly C, Amrein PC, Fathi AT. A phase 1 study of the antibody-drug conjugate brentuximab vedotin with re-induction chemotherapy in patients with Cd30-expressing relapsed/refractory acute myeloid leukemia. Cancer 2020; 1126(6):1264-1273. org/10.1002/cncr.32657.  PMID 31860140