PMID- 12599178
OWN - NLM
STAT- MEDLINE
DCOM- 20040220
LR  - 20091211
IS  - 1552-4949 (Print)
IS  - 1552-4949 (Linking)
VI  - 52
IP  - 1
DP  - 2003 Mar
TI  - Vortex disaggregation for flow cytometry allows direct histologic correlation: a 
      novel approach for small biopsies and inaspirable bone marrows.
PG  - 20-31
AB  - BACKGROUND: Many approaches to obtaining single cells from tissue for flow 
      cytometric immunophenotyping are used; however, these methods result in tissue 
      that is too disrupted for subsequent histologic examination. We introduce a new 
      technique for cell dissociation of hematopoietic malignancies that preserves 
      tissue for histology. This is especially important with small specimens for which 
      this type of correlation is critical. METHODS: Fresh tissue from lymph node, 
      gastrointestinal (GI) tract, skin, and other soft tissue biopsies, in addition to 
      cores of inaspirable bone marrows, were briefly vortexed until the RPMI cell 
      culture medium became cloudy. Larger specimens such as lymph nodes were sectioned 
      before disaggregating, whereas smaller ones were vortexed in toto. Resultant flow 
      cytometric analyses were compared with the histology and, in some cases, the 
      immunohistochemistry (IHC) to determine whether the data were concordant. Cell 
      suspensions of 104 specimens-composed of 48 lymph nodes, 19 bone marrow cores 
      (BMCs), 11 GI biopsies, 11 skin/soft tissue biopsies, and 15 miscellaneous 
      specimens-were prepared via vortex disaggregation. RESULTS: Flow cytometric 
      analysis of 96 specimens (92.3%) showed adequacy of material and diagnostic 
      correlation with the histology and IHC. Of the eight cases (7.7%) that were 
      discordant, seven were attributable to significant specimen fibrosis or necrosis. 
      With respect to tissue type, this method produced diagnostic cell suspensions for 
      most lymph nodes (95.8%), GI biopsies (90.9%), and BMCs (89.5%); however, it was 
      less useful for skin/soft tissue samples (81.8%). CONCLUSIONS: Disaggregation of 
      tissue for flow cytometric analysis by vortexing appears to provide adequate and 
      representative cellular material. This technique is ideal for inaspirable bone 
      marrows and small biopsies where tissue preservation for histology is paramount.
CI  - Published 2003 Wiley-Liss, Inc.
FAU - Vos, Jeffrey A
AU  - Vos JA
AD  - Department of Pathology, Madigan Army Medical Center, Tacoma, Washington, USA. 
      Jeffrey.vos@cen.amedd.army.mil
FAU - Simurdak, Jerry H
AU  - Simurdak JH
FAU - Davis, Brad J
AU  - Davis BJ
FAU - Myers, Jerome B
AU  - Myers JB
FAU - Brissette, Mark D
AU  - Brissette MD
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Cytometry B Clin Cytom
JT  - Cytometry. Part B, Clinical cytometry
JID - 101235690
SB  - IM
MH  - Biopsy
MH  - Bone Marrow Cells/pathology
MH  - Cell Separation/*methods
MH  - *Flow Cytometry
MH  - Humans
MH  - Immunophenotyping
MH  - Leukemia/*pathology
MH  - Lymph Nodes/pathology
MH  - Lymphoma/*pathology
MH  - Palatine Tonsil/pathology
MH  - Retrospective Studies
MH  - Skin/pathology
MH  - Stomach/pathology
EDAT- 2003/02/25 04:00
MHDA- 2004/02/21 05:00
CRDT- 2003/02/25 04:00
PHST- 2003/02/25 04:00 [pubmed]
PHST- 2004/02/21 05:00 [medline]
PHST- 2003/02/25 04:00 [entrez]
AID - 10.1002/cyto.b.10002 [doi]
PST - ppublish
SO  - Cytometry B Clin Cytom. 2003 Mar;52(1):20-31. doi: 10.1002/cyto.b.10002.