Preparation and microscopic visualization of multicolor luminescent immunophosphors.
The preparation of charge-stabilized suspensions of small phosphor particles (0.1-0.3 micron) and their coupling with antibodies to immunoreactive conjugates is described. Phosphor particles consisting of yttriumoxisulfide activated with europium served as a model system in the evaluation of the stabilizing properties of several polycarboxylic acids. The optimal reagents were then applied to other phosphors which differ in spectral characteristics as well as in luminescence lifetime. These phosphors were ground to a size of 0.1-0.3 micron and proteins or other macromolecules were adsorbed to the phosphor particles to prepare conjugates of different physico-chemical properties. A time-resolved microscope, suitable for real time visualization of the time-delayed luminescence of the immunophosphors by the human eye, is described in detail. Since most phosphors require excitation with far UV light, a special fluorescence microscope allowing far UV excitation was developed for conventional visualization of the luminescence emitted by the phosphor. The possibility of multiple color labeling using various phosphor conjugates was demonstrated in a model system consisting of haptenized latex beads.
Beverloo
H B
HB
Department of Cytochemistry and Cytometry, State University of Leiden, The Netherlands.
van Schadewijk
A
A
Bonnet
J
J
van der Geest
R
R
Runia
R
R
Verwoerd
N P
NP
Vrolijk
J
J
Ploem
J S
JS
Tanke
H J
HJ
D000327:Adsorption; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D004867:Equipment Design; D004912:Erythrocytes / Q000648:ultrastructure; D005063:Europium; D005456:Fluorescent Dyes; D006801:Humans; D007150:Immunohistochemistry; D008163:Luminescent Measurements; D008214:Lymphocytes / Q000648:ultrastructure; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008860:Microscopy, Ultraviolet / Q000295:instrumentation / Q000379:methods*; D010316:Particle Size; D011817:Rabbits; D013440:Sulfides / Q000528:radiation effects; D013997:Time Factors; D014466:Ultraviolet Rays; D015019:Yttrium / Q000528:radiation effects
D000954:Antigens, Surface; D005456:Fluorescent Dyes; D013440:Sulfides; C038559:yttrium oxide sulfide; D005063:Europium; D015019:Yttrium