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Analysis of Intracellular Antigens by Flow Cytometry: Methods for Cell Permeabilization to Antibodies

  • Conference paper
Flow Cytometry

Part of the book series: NATO ASI Series ((ASIH,volume 67))

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Abstract

Intracellular molecules have been detected for many years using antibodies and other specific ligands conjugated to fluorescent markers. Cells were prepared by established methods of histochemistry and cytochemistry and were analyzed by fluorescence microscopy. By microscopy not only the presence of an intracellular antigen can be identified but also its localization within the cell. Quantitative analysis, however, suffers from the limitations of microscopy in general, namely (1) the relatively small number of cells of a large population that are analyzed, (2) the slowness and subjectivity of data collection and (3) the restricted sensitivity of the human eye to low-intensity fluorescence.

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References

  • Ahnert-Hilger G, Mach W, Foehr KJ, Gratzl M (1989a) Poration by alpha-toxin and streptolysin O: an approach to analyze intracellular processes. In: Turner JR/ Tartakoff AM (ed) Methods in cell biology, vol 31. Academic Press, San Diego New York Berkeley Boston London Sydney Tokyo Toronto, p 63

    Google Scholar 

  • Ahnert-Hilger G, Bader M-F, Bhakdi S, Gratzl M (1989b) Introduction of macromolecules into bovine adrenal medullary chromatin cell and rat pheochromocytoma cells (PC12) by permeabilization with streptolysin O: inhibitory effect of tetanus toxin on cathecholamine secretion. J Neurochem 52: 1751–1758

    Article  PubMed  CAS  Google Scholar 

  • Anderson P, Blue M-L, O’Brien C, Schlossman SF (1989) Monoclonal antibodies reactive with the T cell receptor zeta chain: production and characterization using a new method. J Immunol 143: 1899–1904

    PubMed  CAS  Google Scholar 

  • Anderson P/ Nagler-Anderson C, O’Brien C, Levine H, Watkins S, Slayter HS, Blue M-L, Schlossman SF (1990) A monoclonal antibody reactive with a 15-kDa cytoplasmic granule-associated protein defines a subpopulation of CD8+ T lymphocytes. J Immunol 144: 574–582

    PubMed  CAS  Google Scholar 

  • Andersson U, Hallden G, Persson U, Hed J, Moeller G, De Ley M (1988) Enumeration of IFN-gamma-producing cells by flow cytometry. J Immunol Methods 112: 139–142

    Article  PubMed  CAS  Google Scholar 

  • Baisch H, Gerdes J (1990) Identification of proliferating cells by Ki-67 antibody. In: Darzynkiewicz Z, Crissman HA (eds) Methods in Cell Biology, vol 33. Academic Press, San Diego New York Boston London Sydney Tokyo Toronto, p 217

    Google Scholar 

  • Bancroft JD (1967) An introduction to histochemical technique, Butterworth, London

    Google Scholar 

  • Bardales RH, Al-Katib AM, Carrato A, Koziner B (1989a) Detection of intracytoplasmic immunoglobulin by flow cytometry in B-cell malignancies. J Histochem Cytochem 37: 83–89

    Article  PubMed  CAS  Google Scholar 

  • Bardales RH, Carrato A, Fleischer M, Schwartz MK, Koziner B (1989b) Detection of terminal deoxynucleotidyl transferase ( TdT) by flow cytometry in leukemic disorders. J Histochem Cytochem 37: 509–513

    Article  PubMed  CAS  Google Scholar 

  • Battifora H, Kopinski M (1986) The influence of protease digestion and duration of fixation on the immunostaining of keratins. A comparison of formalin and ethanol fixation. J Histochem Cytochem 34: 1095–1100

    Article  PubMed  CAS  Google Scholar 

  • Bauer KD (1990) Analysis of proliferation-associated antigens. In: Darzynkiewicz Z, Crissman HA (eds) Methods in Cell Biology, vol 33. Academic Press, San Diego New York Boston London Sydney Tokyo Toronto, p 235

    Google Scholar 

  • Bauer KD, Clevenger CV, Endow RK, Murad T, Epstein AL, Scarpelli DG (1986a) Simultaneous nuclear antigen and DNA content quantitation using paraffin-embedded colonic tissue and multiparameter flow cytometry. Cancer Res 46: 2428–2434

    PubMed  CAS  Google Scholar 

  • Bauer KD, Clevenger CV, Williams TJ, Epstein AL (1986b) Assessment of cell cycle-associated antigen expression using multiparameter flow cytometry and antibody-acridine orange sequential staining. J Histochem Cytochem 34: 245–250

    Article  PubMed  CAS  Google Scholar 

  • Berglund DL, Starkey JR (1989) Isolation of viable tumor cells following introduction of labeled antibody to an intracellular oncogene product using electroporation. J Immunol Methods 125: 79–87

    Article  PubMed  CAS  Google Scholar 

  • Berglund DL, Starkey JR (1991) Introduction of antibody into viable cells using electroporation. Cytometry 12: 64–67

    Article  PubMed  CAS  Google Scholar 

  • Bernstein RL, Browne LH, Yu SC, Williams KL (1988) Detergent treatment of Dictyostelium discoideum cells allows examination of internal cell type-specific antigens by flow cytometry. Cytometry 9: 68–74

    Article  PubMed  CAS  Google Scholar 

  • Bolton WE, Mikulka WR, Healy CG, Schmittling RG, Kenyon NS (1992) Expression of proliferation associated antigens in the cell cycle of synchronized mammalian cells. Cytometry 13: 117–126

    Article  PubMed  CAS  Google Scholar 

  • Bruno S, Crissman HA, Bauer KD, Darzynkiewicz Z (1991) Changes in cell nuclei during S-phase: progressive chromatin condensation and altered expression of the proliferation-associated nuclear proteins Ki-67, cyclin (PCNA), pl05, and p34. Exp Cell Res 196: 99–106

    Article  PubMed  CAS  Google Scholar 

  • Buckingham L, Duncan JL (1983) Approximate dimensions of membrane lesions produced by Streptolysin S and Streptolysin O. Biochim Biophys Acta 729: 115–122

    Article  PubMed  CAS  Google Scholar 

  • Butler TW, Heck LW, Huster WJ, Grossi CE, Barton JC (1988) Assessment of total immunoreactive lactoferrin in hematopoietic cells using flow cytometry. J Immunol Methods 108: 159–170

    Article  PubMed  CAS  Google Scholar 

  • Campana D, Coustan-Smith E, Janossy G (1988) Double and triple staining for studying the proliferative activity of human B and T lymphoid cells. J Immunol Methods 107: 79–88

    Article  PubMed  CAS  Google Scholar 

  • Chakrabarti R, Pfeiffer NE, Wylie DE, Schuster SM (1989a) Incorporation of monoclonal antibodies into cells by osmotic permeabilization. J Biol Chem 264: 8214–8221

    PubMed  CAS  Google Scholar 

  • Chakrabarti R, Wylie DE, Schuster SM (1989b) Transfer of monoclonal antibodies into mammalian cells by electroporation. J Biol Chem 264: 15494–15500

    PubMed  CAS  Google Scholar 

  • Clevenger CV, Bauer KD, Epstein AL (1985) A method for simultaneous nuclear immunofluorescence and DNA content quantitation using monoclonal antibodies and flow cytometry. Cytometry 6: 208–214

    Article  PubMed  CAS  Google Scholar 

  • Czerniak B, Herz F, Wersto RP, Koss LG (1987) Expression of Ha-ras oncogen p21 protein in relation to the cell cycle of cultured human tumor cells. Am J Pathol 126:411–416

    PubMed  CAS  Google Scholar 

  • Darzynkiewicz Z, Staiano-Goico L, Kunicka JE, De Leo AB, Old LJ (1986) p53 content in relation to cell growth and proliferation in murine L1210 leukemia and normal lymphocytes. Leuk Res 10: 1383–1389

    Article  PubMed  CAS  Google Scholar 

  • Dent GA, Leglise MC, Pryzwansky KB, Ross DW (1989) Simultaneous paired analysis by flow cytometry of surface markers, cytoplasmic antigens, or oncogene expression with DNA content. Cytometry 10: 192–198

    Article  PubMed  CAS  Google Scholar 

  • Dolbeare F, Kuo W-L, Beisker W, Vanderlaan, Gray JW (1990) Using monoclonal antibodies in bromodeoxyuridine-DNA analysis. In: Darzynkiewicz Z, Crissman HA (eds) Methods in Cell Biology, vol 33. Academic Press, San Diego New York Boston London Sydney Tokyo Toronto, p 207

    Google Scholar 

  • Downey GP, Chan CK, Grinstein S (1989) Actin assembly in electropermeabilized neutrophils: role of G-proteins. Biochem Biophys Res Commun 164: 700–705

    Article  PubMed  CAS  Google Scholar 

  • Drach J, Gattringer C, Glassl H, Schwarting R, Stein H, Huber H (1989) Simultaneous flow cytometric analysis of surface markers and nuclear Ki-67 antigen in leukemia and lymphoma. Cytometry 10: 743–749

    Article  PubMed  CAS  Google Scholar 

  • Einck L, Bustin M (1984) Functional histone antibody fragments traverse the nuclear envelope. J Cell Biol 98: 205–213

    Article  PubMed  CAS  Google Scholar 

  • Elmendorf S, McSharry J, Laffin J, Fogelman D, Lehman JM (1988) Detection of an early Cytomegalovirus antigen with two-color quantitative flow cytometry. Cytometry 9: 254–260

    Article  PubMed  CAS  Google Scholar 

  • Engelhard HH, Butler AB, Bauer KD (1989) Quantification of the c-myc oncoprotein in human glioblastoma cells and tumor tissue. J Neurosurg 71: 224–232

    Article  PubMed  Google Scholar 

  • Engelhard HH, Krupka JL, Bauer KD (1991) Simultaneous quantification of c-myc oncoprotein, total cellular protein, and DNA content using multiparameter flow cytometry. Cytometry 12: 68–76

    Article  PubMed  CAS  Google Scholar 

  • Epstein RJ, Smith PJ, Watson JV, Waters C, Bleehen NM (1989) Oestrogen potentiates topoisomerase-2-mediated cytotoxicity in an activated subpopulation of human breast cancer cells: implications for cytotoxic drug resistance in solid tumors. Int J Cancer 44: 501–505

    Article  PubMed  CAS  Google Scholar 

  • Fiskum G, Craig SW, Decker GL, Lehninger AL (1980) The cytoskeleton of digitonin-treated rat hepatocytes. Proc Natl Acad Sci USA 77:3430–3434

    Article  PubMed  CAS  Google Scholar 

  • Fronko GE, Long W, Wu B, Papadopoulos T, Henderson EE (1989) Relationship between methylation status and expression of an Epstein-Barr virus ( EBV) capsid antigen gene. Biochem Biophys Res Commun 159: 263–270

    Article  PubMed  CAS  Google Scholar 

  • Garner JG, Colvin RB, Schooley RT (1984) A flow cytometric technique for quantitation of B-cell activation. J Immunol Methods 67: 37–51

    Article  PubMed  CAS  Google Scholar 

  • Giaretti W, Di Vinci A, Geido E, Marsano B, Minks M, Bruno S (1990) Measurement of c-myc protein content and cell cycle kinetics of normal and spontaneously transformed murine mastocytes by bivariate flow cytometry. Cell Tissue Kinet 23: 473–485

    PubMed  CAS  Google Scholar 

  • Goldenthal KL, Hedman K, Chen JW, August JT, Willingham, MC (1985) Postfixation detergent treatment for immunofluorescence suppresses localization of some integral membrane proteins. J Histochem Cytochem 33: 813–820

    Article  PubMed  CAS  Google Scholar 

  • Goolsby C, Gay H, Docherty JJ, Todd P (1988) Flow cytometric detection of Herpes simplex virus type 2 infected and transformed cells by immunoenzymatic and indirect immunofluorescence staining. Cytometry 9: 126–130

    Article  PubMed  CAS  Google Scholar 

  • Gore SD, Kastan MB, Goodman SN, Civin CI (1990) Detection of minimal residual T cell acute lymphoblastic leukemia by flow cytometry. J Immunol Methods 132: 275–286

    Article  PubMed  CAS  Google Scholar 

  • Graham ML, Dalquist KE, Horwitz KB (1989) Simultaneous measurement of progesteron receptors and DNA indices by flow cytometry: analysis of breast cancer cell mixtures and genetic instability of the T47D line. Cancer Res 49: 3943–3949

    PubMed  CAS  Google Scholar 

  • Gratzner HG (1982) Monoclonal antibody to 5-bromo and 5-iododeoxyuridine: a new reagent for detection of DNA replication. Science 218: 474–475

    Article  PubMed  CAS  Google Scholar 

  • Hallden G, Andersson U, Hed J, Johansson SGO (1989) A new membrane permealization method for the detection of intracellular antigens by flow cytometry. J Immunol Methods 124: 103–109

    Article  PubMed  CAS  Google Scholar 

  • Hatfield JM, Hymer WC (1985) Flow cytometric immunofluorescence of rat anterior pituitary cells. Cytometry 6: 137–142

    Article  PubMed  CAS  Google Scholar 

  • Hayden GE, Walker KZ, Miller JFAP, Wotherspoon JS, Raison RL (1988) Simultaneous cytometric analysis for the expression of cytoplasmic and surface antigens in activated T cells. Cytometry 9: 44–51

    Article  PubMed  CAS  Google Scholar 

  • Hjerten S, Johansson K-E (1972) Selective solubilization with Tween-20 of membrane proteins from Acholeplasma laidlawii. Biochim Biophys Acta 288: 312–325

    Article  PubMed  CAS  Google Scholar 

  • Hofsli E, Bakke O, Nonstad U, Espevik T (1989) A flow cytometric and immunofluorescence microscopic study of tumor necrosis factor production and localization in human monocytes. Cell Immunol 122: 405–415

    Article  PubMed  CAS  Google Scholar 

  • Horan M, Horan PK, Williams CA (1975) Quantitative measurement of SV40 T-antigen production. Exp Cell Res 91: 247–252

    Article  PubMed  CAS  Google Scholar 

  • Jacob MC, Favre M, Bensa JC (1991) Membrane cell permeabilization with saponin and multiparametric analysis by flow cytometry. Cytometry 12: 550–558

    Article  PubMed  CAS  Google Scholar 

  • Jacobberger JW, Fogleman D, Lehman JM (1986) Analysis of intracellular antigens by flow cytometry. Cytometry 7: 356–364

    Article  PubMed  CAS  Google Scholar 

  • Kastan MB, Stone KD, Civin CI (1989) Nuclear oncoprotein expression as a function of lineage, differentiation stage, and proliferative status of normal human hematopoietic cells. Blood 74: 1517–1524

    PubMed  CAS  Google Scholar 

  • Kirkhus B, Clausen OPF (1990) Cell kinetics in mouse epidermis studied by bivariate DNA/bromodeoxyuridine and DNA/keratin flow cytometry. Cytometry 11: 253–260

    Article  PubMed  CAS  Google Scholar 

  • Kurki P, Lotz M, Ogata K, Tan EM (1987) Proliferating cell nuclear antigen ( PCNA)/cyclin in activated human T-lymphocytes. J Immunol 138: 4114–4120

    PubMed  CAS  Google Scholar 

  • Kurki P, Ogata K, Tan EM (1988) Monoclonal antibodies to proliferating cell nuclear antigen ( PCNA)/cyclin as probes for proliferating cells by immunofluorescence microscopy and flow cytometry. J Immunol Methods 109: 49–59

    Article  PubMed  CAS  Google Scholar 

  • Labalette-Houache M, Torpier G, Capron A, Dessaint JP (1991) Improved permeabilization procedure for flow cytometric detection of internal antigens. Analysis of interleukin-2 production. J Immunol Methods 138: 143–153

    Article  PubMed  CAS  Google Scholar 

  • Laffin J, Fogleman D, Lehman JM (1989) Correlation of DNA content, p53, T-antigen, and V-antigen in Simian virus 40-infected human diploid cells. Cytometry 10: 205–213

    Article  PubMed  CAS  Google Scholar 

  • Landberg G, Roos G (1991) Antibodies to proliferating cell nuclear antigen as S-phase probes in flow cytometric cell cycle analysis. Cancer Res 51: 4570–4574

    PubMed  CAS  Google Scholar 

  • Larsen JK, Christensen IJ, Christiansen J, Mortensen BT (1991) Washless double staining of unfixed nuclei for flow cytometric analysis of DNA and a nuclear antigen (Ki-67 or bromodeoxyuridine). Cytometry 12: 429–437

    Article  PubMed  CAS  Google Scholar 

  • Lemons R, Forster S, Thoene J (1988) Protein microinjection by protease permeabilization of fibroblasts. Anal Biochem 172: 219–227

    Article  PubMed  CAS  Google Scholar 

  • Levy HP, Schultz RA, Ordonez JV, Cohen MM (1991) Anti-kinetochore staining for single laser, bivariate flow sorting of Indian muntjac chromosomes. Cytometry 12: 695–700

    Article  PubMed  CAS  Google Scholar 

  • Lincoln ST, Bauer KD (1989) Limitations in the measurement of c-myc oncoprotein and other nuclear antigens by flow cytometry. Cytometry 10: 456–462

    Article  PubMed  CAS  Google Scholar 

  • Maeda K, Dixon WT, Jerry LM (1990) Characterization of human melanosome-associated antigen recognized by monoclonal antibody HMSA-2. J Invest Dermatol 94: 221–226

    Article  PubMed  CAS  Google Scholar 

  • Mann GJ, Dyne M, Musgrove EA (1987) Immunofluorescent quantification of ribonucleotide reductase Ml subunit and correlation with DNA content by flow cytometry. Cytometry 8: 509–517

    Article  PubMed  CAS  Google Scholar 

  • Mann KP, Staunton D, Thorley-Lawson DA (1985) Epstein-Barr virus-encoded protein found in plasma membranes of transformed cells. J Virol 55: 710–720

    PubMed  CAS  Google Scholar 

  • McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative, a new fixative for immunoelectron microscopy. J Histochem Cytochem 22: 1077–1083

    Article  PubMed  CAS  Google Scholar 

  • Mercer WE, Nelson D, De Leo AB, Old LJ, Baserga R (1982) Microinjection of monoclonal antibody to protein p53 inhibits serum-induced DNA synthesis in 3T3 cells. Proc Natl Acad Sci USA 79: 6309–6312

    Article  PubMed  CAS  Google Scholar 

  • Miller MR, Ulrich RG, Wang TS-F, Korn D (1985) Monoclonal antibodies against human DNA polymerase-alpha inhibit DNA replication in permeabilized human cells. J Biol Chem 260: 134–138

    PubMed  CAS  Google Scholar 

  • Morgan DO, Roth RA (1987) Acute insulin action requires insulin receptor kinase activity: introduction of an inhibitory monoclonal antibody into mammalian cells blocks the rapid effects of insulin. Proc Natl Acad Sci USA 84: 41–45

    Article  PubMed  CAS  Google Scholar 

  • Morkve O, Laerum OD (1991) Flow cytometric measurment of p53 protein expression and DNA content in paraffin-embedded tissue from bronchial carcinomas. Cytometry 12: 438–444

    Article  PubMed  CAS  Google Scholar 

  • Morris GF, Mathews MB (1989) Regulation of proliferating cell nuclear antigen during the cell cycle. J Biol Chem 264: 13856–13864

    PubMed  CAS  Google Scholar 

  • Nelson KG, Haskill JS, Sloan S, Siegfried JM, Siegal GP, Walton L, Kaufman DG (1987) Flow cytometric analysis of human uterine sarcomas and cell lines. Cancer Res 47: 2814–2820

    PubMed  CAS  Google Scholar 

  • Neubauer A, Serke S, Siegert W, Kroll W, Musch R, Huhn D (1989) A flow cytometric assay for the determination of cell proliferation with a monoclonal antibody directed against DNA-methyltransferase. Br J Haematol 72: 492–496

    Article  PubMed  CAS  Google Scholar 

  • Ojcius DM, Young JD-E (1991) Cytolytic pore-forming proteins and peptides: is there a common structural motif? Trends Biochem Sci 16: 225–229

    Article  PubMed  CAS  Google Scholar 

  • Oud PS, Henderik JBJ, Beck HIM, Veldhuizen JAM, Vooijs G, Herman CJ, Ramaekers FCS (1985) Flow cytometric analysis and sorting of human endometrial cells after immunocytochemical labeling for cytokeratin using a monoclonal antibody. Cytometry 6: 159–164

    Article  PubMed  CAS  Google Scholar 

  • Palutke M, KuKuruga D, Tabaczka P (1987) A flow cytometric method for measuring lymphocyte proliferation directly from tissue culture plates using Ki-67 and propidium iodide. J Immunol Methods 105: 97–105

    Article  PubMed  CAS  Google Scholar 

  • Pattanapanyasat K, Webster HK, Udomsangpetch R, Wanachiwanawin W, Yongvanitchit K (1992) Flow cytometric two-color staining technique for simultaneous determination of human erythrocyte membrane antigen and intracellular malarial DNA. Cytometry 13: 182–187

    Article  PubMed  CAS  Google Scholar 

  • Preis PN, Waldman FM, Frackelton AR, Saya H, Levin VA (1988) Protein- tyrosine kinase activity and pp60 v-src expression in whole cells measured by flow cytometry. Cancer Res 48: 4633–4638

    PubMed  CAS  Google Scholar 

  • Puchtler H, Meloan SN (1985) On the chemistry of formaldehyde fixation and its effects on immunohistological reactions. Histochemistry 82: 201–204

    Article  PubMed  CAS  Google Scholar 

  • Rabbitts PH, Watson JV, Lamond A, Forster A, Stinson MA, Evan G, Fisher W, Atherton E, Shepport R, Rabbitts FH (1985) Metabolism of c-myc gene products: c-myc RNA and protein expression in the cell cycle. EMBO J 4: 2009–2015

    PubMed  CAS  Google Scholar 

  • Rabinovitch PS, Torres RM, Engel D (1986) Simultaneous cell cycle analysis and two-color surface immunofluorescence using 7-amino-actinomycin D and single laser excitation: applications to study of cell activation and the cell cycle of murine LY-1 B cells. J Immunol 136: 2769–2775

    PubMed  CAS  Google Scholar 

  • Reeve JG, Stewart J, Watson JV, Wulfrank D, Twentyman RR, Bleehen NM (1986) Neuron specific enolase expression in carcinoma of the lung. Br J Cancer 53: 519–528

    Article  PubMed  CAS  Google Scholar 

  • Remvikos Y, Laurent-Puig P, Salmon RJ, Frelat G, Dutrillaux B, Thomas G (1990) Simultaneous monitoring of p53 protein and DNA content of colorectal adenocarcinoma by flow cytometry. Int J Cancer 45: 450–456

    Article  PubMed  CAS  Google Scholar 

  • Rice GC, Pennica D, Borree JA, Williams SR (1991) Measurement of transient cDNA expression in mammalian cells using flow cytometric cell analysis and sorting. Cytometry 12: 221–233

    Article  PubMed  CAS  Google Scholar 

  • Roberts P, Jones M, Gale R, Thomas S, Tidman N, Linch D (1989) The c-myc oncogene is regulated independently of differentiation in myeloid cell lines. Leuk Res 13: 651–659

    Article  PubMed  CAS  Google Scholar 

  • Robertson FM, Gilmour SK, Beavis AJ, O’Connel WSM, Conney AH, Huang M-T, Laskin JD, Hietala OA, O’Brien TG (1990) Flow cytometric detection of ornitine decarboxylase activity in epidermal cell subpopulations. Cytometry 11: 832–836

    Article  PubMed  CAS  Google Scholar 

  • Ronot X, Gaillard-Froger B, Hainque B, Adolphe M (1988) In vitro aging of articular chondrocytes identified by analysis of DNA and tubulin content and relationship to cell size and protein content. Cytometry 9:436–440

    Article  Google Scholar 

  • Schimenti KJ, Jacobberger JW (1992) Fixation of mammalian cells for flow cytometric evaluation of DNA content and nuclear immunofluorescence. Cytometry 13: 48–59

    Article  PubMed  CAS  Google Scholar 

  • Schulz I (1990) Permeabilizing cells: some methods and applications for the study of intracellular processes. In: Fleischer S, Fleischer B (eds) Methods in enzymology, vol 192. Academic Press, San Diego New York Boston London Sydney Tokyo Toronto, p 280

    Google Scholar 

  • Schmid I, Uittenbogaart CH, Giorgi JV (1991) A gentle fixation and permeabilization method for combined cell surface and intracellular staining with improved precision in DNA quantification. Cytometry 12: 279–285

    Article  PubMed  CAS  Google Scholar 

  • Schroff RW, Bucana CD, Klein RA, Farrell NM, Morgan AC (1984) Detection of intracytoplasmic antigen by flow cytometry. J Immunol Methods 70: 167–177

    Article  PubMed  CAS  Google Scholar 

  • Schwarting R, Gerdes J, Niehus J, Jaeschke L, Stein H (1986) Determination of the growth fraction in cell suspensions by flow cytometry using the monoclonal antibody Ki-67. J Immunol Methods 90: 65–70

    Article  PubMed  CAS  Google Scholar 

  • Sengupta S, Petsche D, Gelfand EW, Chechik BE (1985) A flow cytometric method for the detection of adenosine deaminase in mononuclear cells. J Immunol Methods 80: 155–162

    Article  PubMed  CAS  Google Scholar 

  • Shapiro HM (1988) Practical flow cytometry, 2nd edn. Alan R Liss, New York

    Google Scholar 

  • Shen D-W, Real FX, De Leo AB, Old LJ, Marks PA, Rifkin RA (1983) Protein p53 and inducer-mediated erythroleukemia cell commitment to terminal cell division. Proc Natl Acad Sci USA 80: 5919–5922

    Article  PubMed  CAS  Google Scholar 

  • Shibata M, Hirono Y, Takahashi M, Kaneda T (1989) Expression of DNA polymerase alpha and Leu3a molecules in growing and saturated cultures of human leukemic cells: phenotype analysis of proliferating cells by flow cytometry. Jap J Cancer Res 80: 1083–1088

    Article  CAS  Google Scholar 

  • Slaper-Cortenbach ICM, Admiraal LG, Kerr JM, van Leeuwen EF, von dem Borne AEGK, Tetteroo PAT (1988). Flow-cytometric detection of terminal deoxynucleotidyl transferase and other intracellular antigens in combination with membrane antigens in acute lymphatic leukemias. Blood 72: 1639–1644

    PubMed  CAS  Google Scholar 

  • Smith PJ, Nakeff A, Watson JV (1985) Flow cytometric detection of changes in the fluorescence emission spectrum of a vital DNA-specific dye in human tumor cells. Exp Cell Res 15 9: 37–46

    Article  CAS  Google Scholar 

  • Stokke P, Erikstein B, Holte H, Funderud F, Steen HB (1991) Cell cycle-specific expression and nuclear binding of DNA polymerase alpha. Molec Cell Biol 11: 3384–3389

    PubMed  CAS  Google Scholar 

  • Thompson WS, Gross RH (1988) Antibodies introduced into living cells with liposomes localize specifically and inhibit specific intracellular processes. Gene Anal Techn 5: 73–79

    Article  CAS  Google Scholar 

  • Turner JR, Tartakoff AM, Berger M (1988) Intracellular degradation of the complement C3b/C4b receptor in the absence of ligand. J Biol Chem 263: 4914–4920

    PubMed  CAS  Google Scholar 

  • Turner JR, Tartakoff AM, Berger M (1989) A flow cytometric method for the quantitative analysis of intracellular and surface membrane antigens. In: Tartakoff AM (ed) Methods in Cell Biology, vol 31. Academic Press, San Diego New York Berkeley Boston London Sydney Tokyo Toronto, p 351

    Google Scholar 

  • van Erp PEJ, Rijzewijk JJ, Boezeman JBM, Leenders J, de Mare S, Schalkwijk J, van de Kerkhof PCM, Ramaekers FCS, Bauer FW (1989) Flow cytometric analysis of epidermal subpopulations from normal and psoriatic skin using monoclonal antibodies against intermediate filaments. Am J Pathol 135: 865–870

    PubMed  Google Scholar 

  • van Roon MA, Aten JA, van Oven CH, Charles R, Lamers W (1989) The initiation of hepatocyte-specific gene expression within embryonic hepatocytes is a stochastic event. Dev Biol 136: 508–516

    Article  PubMed  Google Scholar 

  • Watson JV, Sikora K, Evan GI (1985) A simultaneous flow cytometric assay for c-myc oncoprotein and DNA in nuclei from paraffin embedded material. J Immunol Methods 83: 179–192

    Article  PubMed  CAS  Google Scholar 

  • Weigel PH, Ray DA, Oka JA (1983) Quantitation of intracellular membrane-bound enzymes and receptors in digitonin-permeabilized cells. Anal Biochem 133: 437–449

    Article  PubMed  CAS  Google Scholar 

  • Weltzien HU (1979) Cytolytic and membrane perturbing properties of lysophosphatidylcholine. Biochim Biophys Acta 559: 259–287

    PubMed  CAS  Google Scholar 

  • White E, Spector D, Welch W (1988) Differential distribution of adenovirus E1A proteins and colocalization of E1A with the 70-kilodalton cellular heatshock protein in infected cells. J Virol 62: 4153–4166

    PubMed  CAS  Google Scholar 

  • Wolf ME, Zigmond MJ, Kapatos G (1989) Tyrosine hydroxylase content of residual striatal dopamine nerve terminals following 6-hydroxydopamine administration: a flow cytometric study. J Neurochem 53: 879–885

    Article  PubMed  CAS  Google Scholar 

  • Womack MD, Kendall DA, Macdonald RC (1983) Detergent effects on enzyme activity and solubilization of lipid bilayer membranes. Biochim Biophys Acta 733: 210–215

    Article  PubMed  CAS  Google Scholar 

  • Zarbo RJ, Visscher DW, Crissman JD (1989) 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

    PubMed  CAS  Google Scholar 

  • Zeile G (1980) Intracytoplasmic immunofluorescence in multiple myeloma. Cytometry 1: 37–41

    Article  PubMed  CAS  Google Scholar 

  • Zipf TF, Bryant LD, Koskowich GN, MacGregor SE, Chin L, Johnson H (1984) Enumeration of cytoplasmic u immunoglobulin positive acute lymphoblastic leukemia cells by flow cytometry: comparison with fluorescence microscopy. Cytometry 5: 610–613

    Article  PubMed  CAS  Google Scholar 

  • Zocchi MR, Marelli F, Poggi A (1990) Simultaneous cytofluorometric analysis for the expression of cytoplasmic antigens and DNA content in CD3- human thymocytes. Cytometry 11: 883–887

    Article  PubMed  CAS  Google Scholar 

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  1. Division of Interdepartmental Equipment, The Hebrew University-Hadassah Medical School, P.O.Box 1172, 91010, Jerusalem, Israel

    H. Giloh

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  1. Laboratoire de Cytométrie, Centre National de la Recherche Scientifique, B. P. 3, 94801, Villejuif, France

    Alain Jacquemin-Sablon

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Giloh, H. (1993). Analysis of Intracellular Antigens by Flow Cytometry: Methods for Cell Permeabilization to Antibodies . In: Jacquemin-Sablon, A. (eds) Flow Cytometry. NATO ASI Series, vol 67. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-84616-8_5

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  • DOI: https://doi.org/10.1007/978-3-642-84616-8_5

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-84618-2

  • Online ISBN: 978-3-642-84616-8

  • eBook Packages: Springer Book Archive

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