Biomedical Engineering Reference
In-Depth Information
(4)
Incubate at room temperature for 1 h with gentle shaking.
(5)
Wash with DPBS three times to remove excess reagents.
(6)
Incubate the with Prussian blue staining solution (equal volumes of 20%
HCl and 10% potassium ferrocyanide aqueous solution) for 40 min at room
temperature.
(7)
Wash the cells twice with DPBS buffer.
(8)
Place under the microscope for inspection.
The studies done by the group of Xu at Ocean NanoTech
285
demonstrated the
effectiveness of IO-Ab mediated CTCs capture in fresh human whole blood.
The results shown in
Figure 7.10
A indicated the presence of IO on the cell sur-
face through the Prussian blue staining which was not observed in the cells that
were exposed to IO alone (
Figure 7.10
B).
285
The TEM images shown in
Figure
7.10
C,D also proved the attachment of the IO-Ab on the cell surface even after
the cells were washed multiple times with DPBS.
285
In this study, only 300 of
the CMFDA pre-stained SK-BR3 cells were added to 1 mL whole blood in a
ratio of cancer cells to blood cells (WBCs and RBCs) at about 1:10,000,000.
The IO-Ab caused the formation of IO-Ab + SK-BR3 cells which were resus-
pended in 100 µL DPBS. The CMFDA stained SK-BR3 cells appeared greenish
yellow in color which were bigger in diameter than the WBCs and RBCs as
show in
Figure 7.11
A. An enrichment factor was calculated at 1:10,000,000
based on the number of WBCs and RBCs in the spiked whole blood.
285
The
unique properties of the hydrophilic biocompatible polymer coating on the IO
NPs contributed to the high capture rate in fresh human whole blood. This sys-
tem holds promise for the specific isolation CTCs in fresh human whole blood
that can have the way for early diagnosis and treatment of cancer.
FIGURE 7.11
Separation of SK-BR3 cell that was spiked in female whole blood. (A) SK-BR3
cell pre-stained with CMFDA, (B) isolated CMFDA pre-stained SK-BR3 cells from whole blood,
(C) IO-Ab captured cell from spiked whole blood.
Courtesy of Xu, et al.
285
(For color version of
this figure, the reader is referred to the online version of this topic)
