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ChemComm
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COMMUNICATION
ChemComm
off/on switching properties.23 We believe that the synthetic and SENTAN, JST to K.H., who was also supported by Mochida
DOI: 10.1039/C8CC02451K
availability of unsymmetrical Si-rhodamines opens up new Memorial Foundation for Medical and Pharmaceutical
opportunities for expanding the range of applications of red-to-NIR Research.
fluorescence probes.
(a)
Conflicts of interest
There are no conflicts to declare.
azoSiR640 (Φfl <0.001)
Non-fluorescent
2,6-diMe SiR640 (Φfl = 0.18)
Ex/Em = 637/660 nm
(b)
(d)
Fluorescent
Notes and references
70
60
50
40
30
20
10
0
1000
1
B. N. G. Giepmans, S. R. Adams, M. H. Ellisman and R. Y. Tsien,
Hypoxia
Normoxia
800
600
400
200
0
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2
3
T. Ueno and T. Nagano, Nat. Methods, 2011, 8, 642-645.
R. Weissleder and V. Ntziachristos, Nat. Med., 2003, 9, 123-
*
128.
4
5
6
7
Y. Kushida, T. Nagano and K. Hanaoka, Analyst, 2015, 140
,
,
685-695.
20%
5%
1%
0.1%
0
600 1200 1800 2400 3000 3600
Time (sec)
Oxygen concentration
T. Ikeno, T. Nagano and K. Hanaoka, Chem. Asian J., 2017, 12
(c)
1435-1446.
O2 concentration
Y. Koide, Y. Urano, K. Hanaoka, T. Terai and T. Nagano, ACS
20%
5%
1%
0.1%
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S. Uno, M. Kamiya, T. Yoshihara, K. Sugawara, K. Okabe, M. C.
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50
8
9
H. Sasaki, K. Hanaoka, Y. Urano, T. Terai and T. Nagano, Bioorg.
Med. Chem., 2011, 19, 1072-1078.
30 mm
2
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(e)
kidney
Rao, ACS Chem. Biol., 2010, 5, 1065-1074.
No ligation
16 min
38 min
24 min
46 min
32 min
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kidney
kidney
liver
liver
liver
Ligated (36 min)
54 min
kidney
liver
kidney
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Fig. 4 Far-red to NIR fluorescence probe for hypoxia, azoSiR640. (a)
NIR fluorescence activation of azoSiR640. azoSiR640 is almost non-
fluorescent, but is reduced by bioreductases under hypoxia to
strongly fluorescent 2,6-diMe SiR640. (b) Time-dependent changes
in the fluorescence intensity of azoSiR640 (1 μM) in the presence of
rat liver microsomes (226 μg/3 mL) under hypoxia or normoxia.
Measurements were performed in a potassium phosphate buffer
(0.1 M; pH 7.4) containing DMSO (0.1%) as a cosolvent. As a cofactor
for reductases, NADPH (50 μM) was added after 5 min (arrowhead).
The excitation and emission wavelengths were 640 and 660 nm. (c)
Fluorescence confocal microscopy images of live A549 cells. A549
cells were incubated with azoSiR640 (100 nM) containing DMSO
(0.1%) as a cosolvent at various oxygen concentrations for 6 hr. (d)
Quantitation of fluorescence intensity at ROI of cells under various
oxygen concentrations. n = 4. * indicates p<0.05 by Student’s t-test.
(e) Fluorescence images of mouse. ICR mouse was injected with 100
μM azoSiR640 in 100 μL saline solution containing 1% DMSO as a
cosolvent from an orbital vein. The portal vein and renal vein were
ligated about 35 min after probe injection. Excitation and emission
wavelengths were 635 nm and 700 nm, respectively. Exposure time:
300 ms.
19 T. Myochin, K. Hanaoka, S. Iwaki, T. Ueno, T. Komatsu, T. Terai,
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Nakadate, R. Kojima and Y. Urano, Bioconjugate Chem., 2018,
29, 241-244.
This work was supported in part by grants by JSPS KAKENHI
Grant Numbers 16H00823, 16H05099 and 18H04609 to K.H.,
4 | Chem. Commun., 2018, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
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