Production, fixation, and staining of cells on slides for maximum photometric sensitivity

Robert C. Leif; Patrick M. Harlow; Lidia M. Vallarino

doi:10.1117/12.180792

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21 July 1994 Production, fixation, and staining of cells on slides for maximum photometric sensitivity

Robert C. Leif, Patrick M. Harlow, Lidia M. Vallarino

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Proceedings Volume 2136, Biochemical Diagnostic Instrumentation; (1994) https://doi.org/10.1117/12.180792
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

The need to detect increasingly low levels of antigens or polynucleotides in cells requires improvements in both the preparation and the staining of samples. The combination of centrifugal cytology with the use of glyoxal as cross-linking fixative produces monolayers of cells having minimum background fluorescence. Detection can be further improved by the use of a recently developed type of luminescent tag containing a lanthanide(III) ion as the light- emitting center. These novel tags are macrocyclic complexes functionalized with an isothiocyanate group to allow covalent coupling to a biosubstrate. The Eu(III) complex possesses a set of properties -- water solubility, inertness to metal release over a wide pH range, ligand-sensitized narrow-band luminescence, large Stoke's shift, and long excited-state lifetime -- that provides ease of staining as well as maximum signal with minimum interference from background autofluorescence. Luminescence efficiency studies indicate significant solvent effects.

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Robert C. Leif, Patrick M. Harlow, and Lidia M. Vallarino "Production, fixation, and staining of cells on slides for maximum photometric sensitivity", Proc. SPIE 2136, Biochemical Diagnostic Instrumentation, (21 July 1994); https://doi.org/10.1117/12.180792

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