Quantitative cell bioimaging using gold-nanoshell conjugates and phage antibodies.
The authors describe a quantitative evaluation of the efficacy of cell labeling with plasmon-resonant light-scattering nanoparticles used as contrast agents for dark-field microscopy imaging. The experimental model is based on the biospecific labeling of pig embryo kidney (SPEV) cells with primary phage antibodies, followed by the dark-field microscopic visualization using conjugates of silica/gold nanoshells with secondary rabbit antiphage antibodies. To quantify nanoparticle binding, the authors introduce the labeling-efficacy factor (LEF) which is equal to the ratio of the bound-particle pixels per cell to the total number of pixels occupied by the cell. The LEF is calculated by an imaging-analysis algorithm based on the freely available ImageJ Java-based processing code. In terms of the LEF, a distinct difference was found between intact, nonspecifically labeled, and biospecifically labeled cells.
Khanadeev
Vitaly A
VA
Department of Biophysics, Saratov State University, 83 Ulitsa Astrakhanskaya, Saratov 410012, Russia.
Khlebtsov
Boris N
BN
Staroverov
Sergey A
SA
Vidyasheva
Irina V
IV
Skaptsov
Alexander A
AA
Ileneva
Ekaterina S
ES
Bogatyrev
Vladimir A
VA
Dykman
Lev A
LA
Khlebtsov
Nikolai G
NG
D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000378:metabolism; D017104:Bacteriophage M13 / Q000276:immunology*; D002460:Cell Line; D003624:Darkness; D006046:Gold / Q000737:chemistry*; D007091:Image Processing, Computer-Assisted; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008853:Microscopy / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D057145:Nanoshells / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000378:metabolism; D013194:Staining and Labeling
D000906:Antibodies; D011956:Receptors, Cell Surface; D006046:Gold