Abstract
Many fundamental questions of biology reduce to a common technical problem, how to assess the phenotype of cells in relation to other cells or in relation to morphology. Traditionally, this problem has been approached by classical biochemical analysis, in which heterogeneous tissue is disrupted and proteins or nucleic acid are determined on the extracts. The hope is that if one particular cell type manifests the interesting biology, the overall change in the presence of other cells will be large enough to detect. Microdissection and application of techniques such as polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) have extended the biochemical approach for DNA and RNA to a few cells (1), although quantltation is not precise and the techniques are technically demanding. This approach is not useful for determining phenotypic properties relating to the amount of particular proteins in cells, and ultimately, the behavior of cells is established by their phenotype Flow cytometry has been used to quantify protein biomarkers in individual cells, but the method is limited by requiring large numbers of single cells. Further, it lacks the ability to correlate biochemical analysis with morphology (2).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Vogelstem B., Fearon E., Hamtlton S., Kern S., Prersinger A. C., and Leppert M (1988) Genetic alterations during colorectal tumor development N Engl J Ned 319, 525–532
KOSS L G, Bogdan C., Herz F, and Wersto R. (1989) Flow cytometric measurements of DNA and other cell components in human tumors a critmal appraisal. Hum. Patho1 20, 528–548.
National Research Council (U S. Subcommittee on Biological Markers in Urinary Toxicology) (1995) in Blologlcal Markers in Urmary Toxology. National Academy Press, Washington, DC, pp 1–309.
West S. S. (1970) Biophysical cytochemistry, in Introduction to Quantitative Cytochemistry (anonymous, ed ), Academrc, New York, p 451
Nibbering P. H., LeiJh P C J, and van Furth R (1985) Quantitation of monoclonal antibody binding to individual cells by cytophotometry, in Technzques inImmunocytochemistry, Academic, New York, pp 97–114.
Rao J. Y, Hurst R E, Bales W D, Jones P. L, Bass R. A., Archer L T, and Hemstreet G. P (1990) Cellular f-actin levels as a marker for cellular transformation: relationship to cell division and differentiation Cancer Res 50, 2215–2220.
Rao J. Y, Hemstreet G. P, Hurst R E, Bonner R. B, Jones P L., Mm K. W., and Fradet Y. (1993) Alterations in phenotypic biochemical markers in bladder epithelium during tumongenesis. Proc Natl Acad. Sci. USA 90, 8287–8291.
Bonner R. B, Hemstreet G. P, Fradet Y, Rao J. Y, Min K W, and Hurst R. E (1993) Bladder cancer rusk assessment with quantitative fluorescence image analysts of tumor markers in exfoliated bladder cells Cancer 72, 246l–2469.
Fradet Y, Islam N, Boucher L, Parent-Vaugeols C, and Tardtf M. (1987) Polymorphic expression of a human superficial bladder tumor anttgen defined by mouse monoclonal antibodies Proc Natl. Acad Sci USA 84, 7227–7231
Nakamura N, Hurst R. E, West S. S, Menter J M, Golden J F, Corltss D A., and Jones D. D. (1980) Biophystcal cytochemtcal investigations of mtra-cellular heparin in neoplasttc mast cells J. Hzstochem Cytochem 28, 223–230
Hurst R E, ParmLey R T, Nakamura N., West S S, and Denys F R (1981) Heparan sulfate of AH-130 ascites hepatoma cells a cell-surface glycosaminogly-can not displaced by heparin. J Hzstochem Cytochem 29, 73l–737
Hemstreet G P., West S S, Weems W, Echols C K, McFarland S, Lewin J, and Lmdseth G (1983) Quantitative fluorescence measurements of AO-stained normal and malignant bladder cells lnt. J Cancer 31, 577–585
ParmLey R T, Hurst R. E, Takagt M, Spicer S S, and Austin R L (1983) Glycosammoglycans in human neutrophils and leukemic myeloblasts. ultrastruc-tural, cytochemical, mnnunologtc, and btochemtcal charactertzatton Blood 61, 257–266
Bass R. A., Hemstreet G P, Honker N. A, Hurst R E, and Doggett R S (1987) DNA cytometry and cytology by quantitative fluorescence image analysts in symptomatic bladder cancer patients int J Cancer 40, 698–705.
West S. S., Hemstreet G P, Hurst R. E., Bass R. A., Doggett R S., and Schulte P A (1987) Detection of DNA aneuplotdy by quantitative fluorescence image analysis potential in screening for occupational bladder cancer, in Blologxal Monitorzng of Exposure to Chemrcals (Dtllon K and Ho M, eds ), Wtley, NewYork, pp 327–341
McGowan P., Hurst R E, Bass R E., Hemstreet G. P., and Pastier R (1988) Equihbrium binding of Hoechst 33258 and Hoechst 33342 fluorochromes with rat colorectal cells. J Hzstochem Cytochem 36, 757–762.
Rao J. Y., Hemstreet G. P, Hurst R E., Bonner R B, Min K W., and Jones P. L. (1991) Cellular F-actin levels as a marker for cellular transformatton correlation with bladder cancer rusk Cancer Res 51, 2762–2767.
Hemstreet G P., 3d, Rao J. Y, Hurst R E, Bonner R B, Jones P L, Valdya A. M., Fradet Y, Moon R C, and Kelloff G J (1992) Intermediate endpoint biomarkers for chemoprevention J Cell Biochem 161(Suppl.), 93–110
Bonner R. B., Ltebert M., Hurst R E., Grossman H. B., Bane B L, and Hemstreet G P (1996) Marker network for bladder cancer characterlzatton of the DD23 tumor-associated antigen for bladder cancer detection and recurrence monitoring Cancer Epldemrol Blomarkers Prev. 5, 97l–978.
Hemstreet G. P., Bonner R. B., Hurst R. E., and O’Dowd G. A (1996) Cytology of Bladder Cancer, in Comprehenswe Textbook of Genztourznary Oncology (Vogelzang N. J., Scardmo P T, Shipley W U, and Coffey D S, eds ), Williams and Wilkins, Baltimore, MD, pp 338–350
Hemstreet G, Hurst R, and Bonner R. (1998) Selection and development of biomarkers for bladder cancer, in Methods in Molecular Medicine, Humana, Totowa, NJ, pp 37–60
Rao J. Y., Bonner R B., Hurst R. E, Qlu W. R., Rezmkoff C A., and Hemstreet G P. (1997) Quantrtattve changes in cytoskeletal and nuclear actin levels during cellular transformatton lnt J Cancer 70, 423–429
Kremk K. D, Kephart G M., Offord K P., Dunnette S L., and Gletch G. J (1989) Compartson of anttfading agents used in mnnunofluorescence. J Zmmunol Meth 117, 91–97.
Brigah D J., Budgeon L. R, Unger E R., Koebler D., Cuomo C, Kennedy T., and Perdoma J. M (1988) Immunocytochemtstry is automated development of a robottc workstation based upon the caprllary actton princrple J Histotechnol 11, 165–183
Jones P L, O’Hare C, Bass R A, Rao J. Y, Hemstreet G P, and Hurst R E (1990) Quantitative immunofluorescence, anti-ras p21 antibody spechlty and cellular oncoprotem levels Blochem Bzophys Res Commun. 167, 464–470.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Humana Press Inc, Totowa, NJ
About this protocol
Cite this protocol
Bonner, R.B., Hurst, R.E., Rao, J.Y., Hemstreet, G.P. (1998). Instrumentation, Accuracy, and Quality Control Issues in Development of Quantitative Fluorescence-Image Analysis (QFIA). In: Hanausek, M., Walaszek, Z. (eds) Tumor Marker Protocols. Methods in Molecular Medicine™, vol 14. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-380-5:181
Download citation
DOI: https://doi.org/10.1385/0-89603-380-5:181
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-380-1
Online ISBN: 978-1-59259-598-3
eBook Packages: Springer Protocols