Application of quantum dots as probes for correlative fluorescence, conventional, and energy-filtered transmission electron microscopy.
Luminescent semiconductor quantum dots (QDs) are a new class of fluorescent label with wide-ranging applications for cell imaging. The electron density and elemental composition of these materials permit the extension of their use as probes in conventional electron microscopy (TEM) and energy-filtered TEM (EFTEM). Here we illustrate the feasibility of using streptavidin-conjugated QDs as TEM tags by labeling a nuclear protein on cell sections and obtaining correlative fluorescence and TEM data. We also show that QD probes can be employed in conjunction with immunogold for co-localization of proteins at the ultrastructural level. Furthermore, by obtaining cadmium elemental maps of CdSe/ZnS QDs distributed on a nuclear structure, we demonstrate the potential of QDs for co-localization of multiple proteins when used in combination with EFTEM.
Nisman
Rozalia
R
Programme in Cell Biology, The Hospital for Sick Children, Toronto, Ontario, Canada.
Dellaire
Graham
G
Ren
Ying
Y
Li
Ren
R
Bazett-Jones
David P
DP
D045744:Cell Line, Tumor; D005456:Fluorescent Dyes; D006801:Humans; D007150:Immunohistochemistry; D008854:Microscopy, Electron / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009363:Neoplasm Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D000071156:Promyelocytic Leukemia Protein; D045663:Quantum Dots; D014157:Transcription Factors / Q000378:metabolism; D025521:Tumor Suppressor Proteins
D005456:Fluorescent Dyes; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human