%0 Journal Article
%A Xu, Jingjing
%A Jiang, Depeng
%A Qin, Yanling
%A Xia, Juan
%A Jiang, Dechen
%A Chen, Hong-Yuan
%D 2017
%T C3N4 Nanosheet Modified Microwell
Array with Enhanced Electrochemiluminescence for Total Analysis of
Cholesterol at Single Cells
%U https://acs.figshare.com/articles/journal_contribution/C_sub_3_sub_N_sub_4_sub_Nanosheet_Modified_Microwell_Array_with_Enhanced_Electrochemiluminescence_for_Total_Analysis_of_Cholesterol_at_Single_Cells/4639765
%R 10.1021/acs.analchem.6b04635.s001
%2 https://acs.figshare.com/ndownloader/files/7554244
%K hydrogen peroxide
%K microwell array
%K intracellular cholesterol
%K intracellular cholesterol storage
%K g-C 3 N 4 nanosheet
%K C 3 N 4 Nanosheet Modified Microwell Array
%K ACAT
%K cell cholesterol analysis
%K ECL
%K intracellular cholesterol pools
%K membrane cholesterol
%K cholesterol oxidase
%K luminescence
%X Here, a g-C3N4 nanosheet modified microwell
array providing enhanced electrochemiluminescence (ECL) and better
visible sensitivity was prepared
to simultaneously analyze total (membrane and intracellular) cholesterol
at single cells. The detection limit for ECL visualization of hydrogen
peroxide at microwell array was improved to be 500 nM that guaranteed
the detection of low concentration cholesterol at single cells in
parallel. To achieve single cell cholesterol analysis, the individual
cells cultured at the microwell array were exposed to cholesterol
oxidase generating hydrogen peroxide for luminescence analysis of
membrane cholesterol, and then treated with triton X-100, cholesterol
esterase, and cholesterol oxidase to produce hydrogen peroxide from
intracellular cholesterol for luminescence determination. The observation
of the luminescence spots at microwells in these two steps confirmed
the codetection of membrane and intracellular cholesterol at single
cells. The inhibition of intracellular acyl-coA/cholesterol acyltransferase
(ACAT) resulted in less intracellular cholesterol storage (less luminescence)
and more membrane cholesterol (more luminescence). The correlation
of the luminescence intensity with the amount of cholesterol confirmed
that our assay could simultaneously monitor membrane and intracellular
cholesterol pools at different cellular states, which should offer
more information for the study of cholesterol-related pathways at
single cells.
%I ACS Publications