The application of specific and proven guidelines is required for flow cytometric nuclear DNA analysis, which can provide precise information on genomic quantitative modifications and the proliferative activity of tumour tissues, clinically useful for prognosis and monitoring of therapy and recurrencies. The authors review the various protocol phases to be followed for a correct analysis and discuss the main problems that can be encountered in the procedural steps and in interpreting the histograms.
KrissJ.P.ReveszL.: The distribution and fate of bromodeoxyuridine and bromodeoxycytidine in the mouse and rat. Cancer Res., 22: 254–265, 1961.
2.
HiddemannW.: Convention on nomenclature for DNA cytometry. Cancer Genet. Cytogenet., 13: 181–183, 1984.
3.
MerkelD.E.DresslerL.G.McGuireW.L.: Flow cytometry, cellular DNA content and prognosis in human malignancy. J. Clin. Oncol., 8: 1690–1703, 1987.
4.
CoonJ.S.LandayA.L.WeinsteinR.S.: Advances in flowcytometry for diagnostic pathology. Lab. Invest., 57: 453–479, 1987.
5.
SalomonA.BorrowJ.GoddardA.D.: Chromosome aberrations and cancer. Science, 254: 1153–1160, 1991.
6.
GrayJ.W.PinkelD.: Molecular cytogenetics in human cancer diagnosis. Cancer, 69: 1536–1542, 1992.
7.
MauroF.: Flow cytometry as a tool for the prognostic assessment of human neoplasia. Int. J. Radiat. Oncol. Biol. Phys., 12: 625–636, 1986.
8.
WalterM.A.PetersG.E.PeiperS.C.: Predicting radio-resistance in early glottic squamous cell carcinoma by DNA content. Ann. Otol. Rhinol. Laringol., 100: 523–526, 1991.
9.
FriedlanderM.L.HedleyD.W.TaylorI.W.: Clinical and biological significance of aneuploidy in human tumours. J. Clin. Pathol., 37: 961–974, 1984.
10.
VindelovL.: Flow cytometric analysis of nuclear DNA in cells from solid tumours and cell suspensions. Virchows Archiv. B, Cell Pathol., 24: 227–242, 1977.
11.
CrissmanH.A.SteinkampJ.A.: Cytochemical techniques for multivariate analysis of DNA and other cellular constituents. In Flow Cytometry and Sorting, Edited by MelamedM.R.LindmoT.MendelsohnM.L., Wiley-Liss Inc., pp. 227–248, 1990.
12.
VindelovL.L.ChristensenI.J.: A review of techniques and results obtained in one laboratory by an integrated system of methods designed for routine clinical flow cytometric DNA analysis. Cytometry, 11: 753–770, 1990.
VindelovL.L.ChristensenI.J.: Some methods and applications of flow cytometric DNA analysis in clinical and experimental oncology. Eur. J. Haematol. Suppl. 48, 42: 69–76, 1989.
15.
AzzolinaL.S.StevanoniG.TridenteG.: DNA analysis of stimulated lymphocytes by automatic sampling for flow cytometry. Citometry, 9: 508–511, 1988.
16.
FranzolinN.BianchiG.MalassiniG.NovellaG.AzzolinaL.S.ComunaleL.MobilioG.: Valutazione comparativa tra citologia e analisi citofluorimetrica del DNA nucleare del liquido di lavaggio nelle neoplasie vescicali. Acta Urol. Ital., 4 (suppl. 1): 497–499, 1989.
17.
VindelovL.L.ChristensenI.J.KeidingN.Spang-ThomsenM.NissenN.I.: Long term storage of samples for flow cytometric DNA analysis. Cytometry, 3: 317–322, 1983.
18.
LowryJ.RogersK.PercivalR.C.DayC.PotterC.W.UnderwoodJ.C.E.: Flow cytometric DNA analysis of breast carcinomas: a comparative study of fresh, frozen and fixed- embedded breast. Surg. Res. Comm., 2: 27–37, 1987.
19.
LowryJ.DayC.GodsonO.PotterC.: Optimal fixation of cell suspensions for flow citometry. Flow Cytom. Standard. Forum, 3: 1–4, 1991.
20.
HedleyD.W.FriedlanderM.L.TaylorJ.W.RuggCA.MusgroveE.A.: Method for analysis of cellular DNA content of paraffin-embedded pathological material using flow cytometry. J. Histochem. Cytochem., 31: 1333–1355, 1983.
21.
van Driel-KulkerA.M.J.MeskerW.E.van VelzelI.: Preparation of monolayer smears from paraffin-embedded tissue for image cytometry. Citometry, 6: 268–272, 1985.
22.
PetersenS.E.: Accuracy and reliability of flow cytometric DNA analysis using a simple one-step Ethidium Bromide staining protocol. Citometry, 7: 301–306, 1986.
23.
MobilioG.TribukaitB.FranzolinN.AzzolinaL.S.TridenteG.BeltramiP.CurtiP.P.: Prognosis of superficial bladder tumours as studied by DNA flow cytometry. A report on a double blind study of 120 patients. J. Exp. Clin. Cancer Res., 7: 243–249, 1988.
24.
Arndt-JovinD.JovinT.M.: Analysis and sorting of living cells according to deoxyribonucleic acid content. J. Histochem. Cytochem., 25: 585–589, 1977.
25.
DolbeareF.A.GratznerH.G.PallaviciniM.G.GrayJ.W.: Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc. Natl. Ac. Sci. USA, 80: 5573–5577, 1983.
26.
RiccardiA.DanovaM.WilsonG.UcciG.DoermerP.MazziniG.BrugnatelliS.GirinoM.McNallyN.J.AscariE.: Cell kinetics in human malignancies studied with in vivo administration of bromodeoxyuridine and flow cytometry. Cancer Res., 48: 6238–6245, 1988.
27.
VindelovL.L.ChristensenI.J.NissenN.I.: A detergent-trypsin method for the preparation of nuclei for flow cytometric DNA analysis. Cytometry, 3: 323–327, 1983.
28.
VindelovL.L.ChristensenI.J.JensenG.NissenN.I.: Limits of detection of nuclear DNA abnormalities by flow cytometric DNA analysis. Results obtained by a set of methods for sample storage, staining and internal stardardization. Cytometry, 3: 332–339, 1983.
29.
NowellP.C.: Molecular events in tumour development. New Engl. J. Med., 319: 575–577, 1988.
30.
GiarettiW.: A model on the origin and evolution of DNA aneuploidy. Int. J. Oncol., 2: 165–171, 1993.
31.
CowanD.F.WuB.YoungG.P.KhannaO.P.: Correlation of histological, cytological and flow cytometric findings in neoplastic and nonneoplastic lesions of the bladder. J. Urol., 138: 753–757, 1987.
32.
ZarboR.J.VisscherD.W.CrissmanJ.D.: Two color multiparametric method for flow cytometric DNA analysis of carcinomas using staining for cytokeratin and leukocyte-common antigen. Anal. Quant. Cytol. Histol., 11: 391–402, 1989.
33.
SussiP.L.: Analisi citofluorimetrica del DNA nucleare nello struma nodulare. Chir. It., 43: 131–137, 1992.
34.
FranzolinN.: Il ruolo dell'analisi del DNA mediante citometria a flusso nell'identificare i falsi positivi citologici da flogosi. Acta Urol. It., 6 (Suppl. 1): 505–507, 1991.
35.
TraskB.PinkelD.: Fluorescence in situ hybridization with DNA probes. In Flow Cytometry, Edited by DarzynkiewiczZ.CrissmanH.A., Academic Press, Methods in Cell Biology vol. 33, chapt. 37, 383–400, 1990.