Development and application of quantum dots for immunocytochemistry of human erythrocytes.
Recent developments in quantum dot technology have resulted in the introduction of new fluorescence immunocytochemical probes. In contrast to organic fluorophores, which are not photostable, the high quantum yield and remarkable photostability of quantum dots solve major problems associated with immunocytochemical studies of erythrocytes. We report here the first application of quantum dots to immunocytochemical studies of human erythrocytes capable of being used in high-magnification, three-dimensional erythrocyte reconstruction techniques. The procedure consists of stabilizing human erythrocytes with a homofunctional imidoester cross-linker to minimize fixative-induced autofluorescence followed by reacting with a quantum dot - monoclonal antibody complex to label band 3 protein. Our new procedure clearly showed a non-homogeneous, raft-like distribution of band 3 protein in the erythrocyte membrane. We also demonstrate the applicability of our technique to studies of erythrocyte membrane modifications occurring during the invasion of a malaria parasite.
Tokumasu
F
F
Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0425, USA.
Dvorak
J
J
D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism; D000913:Antibodies, Protozoan / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism* / Q000469:parasitology*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D018796:Immunoconjugates; D007150:Immunohistochemistry / Q000379:methods*; D016778:Malaria, Falciparum / Q000175:diagnosis* / Q000469:parasitology; D010963:Plasmodium falciparum; D012666:Semiconductors
D001457:Anion Exchange Protein 1, Erythrocyte; D000913:Antibodies, Protozoan; D005456:Fluorescent Dyes; D018796:Immunoconjugates