How to cite this article: Brincat P, Degaetano J, Donaldson C. Evaluation of a Method Allowing Preservation of Fresh Lymph Nodes for Flow Cytometric Immunophenotyping. Cytometry Part B 2012; 82B: 245–251.
Evaluation of a method allowing preservation of fresh lymph nodes for flow cytometric immunophenotyping†
Article first published online: 26 APR 2012
Copyright © 2012 International Clinical Cytometry Society
Cytometry Part B: Clinical Cytometry
Volume 82B, Issue 4, pages 245–251, July 2012
How to Cite
Brincat, P., Degaetano, J. and Donaldson, C. (2012), Evaluation of a method allowing preservation of fresh lymph nodes for flow cytometric immunophenotyping. Cytometry, 82B: 245–251. doi: 10.1002/cyto.b.21021
- Issue published online: 21 JUN 2012
- Article first published online: 26 APR 2012
- Accepted manuscript online: 12 APR 2012 11:08AM EST
- Manuscript Accepted: 20 MAR 2012
- Manuscript Revised: 1 MAR 2012
- Manuscript Received: 19 OCT 2011
- flow cytometric immunophenotyping;
- haematolymphoid neoplasia;
- lymph nodes;
Flow cytometric immunophenotyping (FCI) of lymph nodes (LN) requires fresh unfixed tissue, with analysis being carried out within few hours post surgery. This study evaluated a novel method for fresh LN preservation, in order to allow histomorphology-based FCI.
This study was carried out prospectively on 30 LN with suspected involvement by haematolymphoid neoplasms (HLN). FCI was performed on each fresh and post cryopreserved LN cell suspension. Percentage positivities (PP) and mean fluorescent intensities (MFI) were calculated on both preparations for a combination of T and B-lymphoid antigens together with viability.
The cryopreservation method applied in this study did not affect significantly PP and had minor impact on MFI of the tested antigens. Overall, there was minimal decrease in PP and MFI on the cryopreserved cells when compared with fresh cells, for most antigens with only a mild increase in apoptotic cells. However, these changes were not diagnostically significant, since both reactive processes and HLN present within analyzed LN could be identified and differentiated. Viability was more than 75% for all cryopreserved LN composed of haematolymphoid cells.
The method presented in this study confers the possibility of storing fresh LN biopsies for later FCI, thus allowing a morphology-based immunophenotypic approach. This would allow a more sensitive, specific, and cost-effective management of LN specimens, whilst maintaining the important benefits provided by FCI. © 2012 International Clinical Cytometry Society