Abstract
We have developed a model system based on the immobilization of microdroplets (0.1 μl volumes) of antigens or of target (sense) oligonucleotides onto aminoalkylsilane-coated glass slides. Oligopeptide antigens need to be vapour-fixed in order to achieve efficient immobilization, while oligonucleotides do not require fixation. Protein antigens, exemplified by rabbit immunoglobulin, may be subjected to liquid fixation. The glass slide models are optically translucent and useful both for in situ hybridization and immunocytochemistry. The slides are compatible with detection systems of both light and fluorescence microscopy and permit measurement of staining intensities by microfluorimetry or computerized microdensitometry. The model systems can be used for comparisons of method sensitivities, for characterizing antibody and probe sensitivities and cross-reactivities, and as internal standards or quality controls for immunocytochemistry and in situ hybridization.
Similar content being viewed by others
References
Brandtzaeg P (1972) Evaluation of immunofluorescence with artificial sections of selected antigenicity. Immunology 22: 177–183
Capel PJA (1975) The defined antigen substrate spheres (DASS) system and some of its applications. Ann NY Acad Sci 254: 108–118
Fuller PJ, Stone DL, Brand SJ (1987) Molecular cloning and sequencing of a rat preprogastrin complementary deoxyribonucleic acid. Mol Endocrinol 1: 306–311
Knapp W, Polem JS (1974) Microfluorometry of antigen-antibody interactions in immunofluorescence using the defined antigen substrate spheres (DASS) system. Sensitivity, specificity and variables of the method. J Immunol Methods 5: 259–273
Larsson L-I (1981a) A novel immunocytochemical model system for specificity and sensitivity screening of antisera against multiple antigens. J Histochem Cytochem 29: 408–410
Larsson L-I (1981b) Adrenocorticotropin-like and α-melanotropinlike peptides in a subpopulation of human gastrin cell granules: bioassay, immunoassay and immunocytochemical evidence. Proc Natl Acad Sci USA 78: 2990–2994
Larsson L-I (1988) Immunocytochemistry, theory and practice. CRC Press, Boca Raton, Florida
Larsson L-I, Hougaard DM (1993) Sensitive detection of rat gastrin mRNA by in situ hybridization with chemically biotinylated oligodeoxynucleotides: validation, quantitation and double-staining studies. J Histochem Cytochem 41: 157–163
Larsson L-I, Rehfeld JF (1981) Pituitary gastrins occur in corticotrophs and melanotrophs. Science 213: 768–770
Larsson L-I, Stengaard-Pedersen K (1981) Enkephalin/endorphin-related peptides in antropyloric gastrin cells. J Histochem Cytochem 29: 1088–1098
Larsson L-I, Stengaard-Pedersen K (1982) Immunocytochemical and ultrastructural differentiation between met-enkephalin-, leu-enkephalin-, and met/leu-enkephalin-immunoreactive neurons of feline gut. J Neurosci 2: 861–878
Larsson L-I, Christensen T, Dalbøge H (1988) Detection of proopiomelanocortin mRNA by in situ hybridization using a biotinylated oligodeoxynucleotide probe and avidin-alkaline phosphatase histochemistry. Histochemistry 89: 109–116
Larsson L-I, Traasdahl B, Hougaard DM (1991) Quantitative nonradioactive in situ hybridization. Model studies and studies on pituitary proopiomelanocortin cells after adrenalectomy. Histochemistry 95: 209–215
Leary JJ, Brigati DJ, Ward D (1983) Rapid and sensitivie colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: bioblots. Proc Natl Acad Sci USA 80: 4045–4049
Maples JA (1985) A method for the covalent attachment of cells to glass slides for use in immunohistochemical assays. Am J Clin Pathol 93: 356–363
Millar DA, Williams ED (1982) A step-wedge standard for the quantification of immunoperoxidase techniques. Histochem J 14: 609–620
Nabors LB, Songu-Mize E, Mize RR (1988) Quantitative immunocytochemistry using an image analyzer. II. Concentration standards for transmitter immunocytochemistry. J Neurosci Methods 26: 25–34
Nairn RC, Rolland JM, Ward HA, Matthews N, Chalmers PJ (1975) Immunofluorescence in cancer investigation and research. Ann NY Acad Sci 254: 523–527
Ottersen OP (1987) Postembedding light- and electron microscopic immunocytochemistry of amini acids: description of a new model system allowing identical conditions for specificity testing and tissue processing. Exp Brain Res 69: 167–174
Posthuma G, Slot JW, Geuze HJ (1987) Usefulness of the immunogold technique in quantitation of a soluble protein in ultrathin sections. J Histochem Cytochem 35: 405–410
Retnrop M, Knapp B, Winter H, Schweizer J (1986) Aminoalkylsilane-treated glass slides as support for in situ hybridization of keratin cDNAs to frozen tissue sections under varying fixation and pretreatment conditions. Histochem J 18: 271–276
Robinson PJ, Dunnill P, Lilly MD (1971) Porous glass as a solid support for immobilization or affinity chromatography of enzymes. Biochim Biophys Acta 242: 659–661
Streefkerk JG, van der Ploeg M, van Duijn P (1975) Agarose beads as matrices for proteins in cytophotometric investigations of immunohistoperoxidase procedures. J Histochem Cytochem 23: 243–250
Towbin H, Gordon J (1984) Immunoblotting and dot immunobinding. Current status and outlook. J Immunol Methods 72: 313–340
van Prooijen-Knegt AC, Raap AK, van der Burg MJM, Vrolijk J, van der Ploeg M (1982) Spreading and staining of human metaphase chromosomes on aminoalkylsilane-treated glass slides. Histochem J 14: 333–344
Weetall HH, Filbert AM (1974) Porous glass for affinity chromatography applications. Methods Enzymol 34: 59–73
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Larsson, LI., Hougard, D.M. Glass slide models for immunocytochemistry and in situ hybridization. Histochemistry 101, 325–331 (1994). https://doi.org/10.1007/BF00268993
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00268993