Time-lapse fluorescence imaging and quantitative single cell and endosomal analysis of peritoneal macrophages using fluorescent organosilica nanoparticles.
Fluorescent thiol-organosilica nanoparticles with 100 nm diameter (F-thiol-OS-100) were applied for time-lapse fluorescence imaging. The evaluation of F-thiol-OS-100 for quantitative analysis demonstrated great advantages as compared with quantum dots and organic fluorescent dye. Time-lapse fluorescence imaging of mouse peritoneal macrophages using F-thiol-OS-100 clearly demonstrated cellular uptake, and single cell analysis showed various patterns of uptake kinetics that could be quantitatively evaluated. We also performed quantitative analysis of endosomal uptake and movements in single cells. A correlation between morphologic findings and endosomal uptake and movement over time was also observed and analyzed quantitatively. The F-thiol-OS-100 showed high potential as a new fluorescence marker for time-lapse fluorescence imaging and quantitative single cell functional analysis for nanomedicine development.


BACKGROUND
In this study the authors report on 100 nm thiol-organosilica nanoparticles as time-lapse flurescent markers. F-thiol-OS-100 proved to be superior to quantum dots and organic flurescent dyes, and enabled quantitative single cell functional analysis.
Department of Anatomy and Cell Biology, The University of Tokushima Graduate School, Tokushima, Japan. michy@basic.med.tokushima-u.ac.jp
Michihiro
M
Nakamura
Kazunori
K
Miyamoto
Koichiro
K
Hayashi
Aziz
A
Awaad
Masahito
M
Ochiai
Kazunori
K
Ishimura
D000818:Animals; D002460:Cell Line; D011992:Endosomes / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D053758:Nanoparticles / Q000032:analysis* / Q000648:ultrastructure; D061848:Optical Imaging / Q000379:methods*; D017646:Organosilicon Compounds / Q000032:analysis* / Q000378:metabolism; D045663:Quantum Dots; D059010:Single-Cell Analysis / Q000379:methods*; D013438:Sulfhydryl Compounds / Q000032:analysis
D005456:Fluorescent Dyes; D017646:Organosilicon Compounds; D013438:Sulfhydryl Compounds