Time-resolved fluorescence imaging for quantitative histochemistry using lanthanide chelates in nanoparticles and conjugated to monoclonal antibodies

Tissue and cell examinations have a potential to produce extremely valuable information about antigen quantities in samples. Using currently available methods, a truly quantitative analysis is nearly impossible. We have previ ously shown that immunohistochemical (IHC) detection of prostate-specific a ntigen and human glandular kallikrein from prostatic tissue, together with time-resolved fluorescence imaging (TRFI), is a suitable method for obtaini ng quantitative data from biological samples and that the signal response i s linear. In this paper we show that Eu-chelate containing particles in the nanometer range are suitable labels for quantitative IHC. Even single nano particle molecules can be detected by TRFI and the signals measured can be readily quantitated. The signal intensity correlates very well with the amo unt of bound label, and the use of nanoparticles could markedly improve the sensitivity of quantitative IHC methods. TRFI provides a powerful tool for providing quantitative data about antigens or transcripts in tissue sectio ns or cultured cells. It is also of major importance in standardization and optimization of protocols for fixation and tissue preparation, including a ntigen retrieval methods. Copyright (C) 2000 John Wiley & Sons, Ltd.