A bodipy based dual functional probe for the detection of hydrogen sulfide and H2S induced apoptosis in cellular systems

Article abstract of DOI:10.1039/c5cc02984h

A bodipy based dual functional probe 1 has been designed and synthesized, which selectively detects H₂S as well as monitors H₂S induced apoptosis in cells.

Full text of DOI:10.1039/c5cc02984h

View Article Online
View Journal
ChemComm
Accepted Manuscript
This article can be cited before page numbers have been issued, to do this please use: N. Gupta, S. I.
Reja, V. Bhalla, M. Gupta, G. kaur and M. Kumar, Chem. Commun., 2015, DOI: 10.1039/C5CC02984H.
This is an Accepted Manuscript, which has been through the
Royal Society of Chemistry peer review process and has been
accepted for publication.
Accepted Manuscripts are published online shortly after
acceptance, before technical editing, formatting and proof reading.
Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available.
You can find more information about Accepted Manuscripts in the
Information for Authors.
Please note that technical editing may introduce minor changes
to the text and/or graphics, which may alter content. The journal’s
standard Terms & Conditions and the Ethical guidelines still
apply. In no event shall the Royal Society of Chemistry be held
responsible for any errors or omissions in this Accepted Manuscript
or any consequences arising from the use of any information it
contains.
www.rsc.org/chemcomm

Page 1 of 4
ChemComm
View Article Online
DOI: 10.1039/C5CC02984H
Journal Name
RSCPublishing
COMMUNICATION
A bodipy based dual functional probe for the
detection of hydrogen sulfide and H2S induced
apoptosis in cellular system
Neha Gupta,a Shahi Imam Reja,a Vandana Bhalla,a Muskan Gupta,b Gurcharan
Kaurb and Manoj Kumar*a
Cite this: DOI:
10.1039/x0xx00000xReceived 00th
January 2012,
Accepted 00th January 2012DOI:
10.1039/x0xx00000x
www.rsc.org/
product will act as a fluorescence molecular rotor (FMR) since
it will have donorꢀπꢀspacerꢀacceptor arrangement,8c,9 a preꢀ
requisite for a molecule to behave as FMR. To our pleasure, the
A bodipy based dual functional probe 1 has been designed
and synthesized which selectively detects H2S as well as
monitors H2S induced apoptosis in cells.
reduced product
2 responded to changes in medium viscosity
Hydrogen sulfide (H2S), being a gaseous signalling molecule is
involved in a wide range of physiological and pathological
responses. The physiological concentration of H2S ranges from
nano to milliꢀmolar levels.1 Once this concentration level of
H2S is disturbed, it leads to various diseases like chronic kidney
disease, liver cirrhosis, Down’s syndrome and Alzheimer’s
disease.2 H2S is also a wellꢀknown apoptosis inducer3 and
activates the key apoptotic enzymes caspaseꢀ9, caspaseꢀ8, and
caspaseꢀ33cꢀ4 and initiates cell death by causing nuclear
condensation and cytoplasmic shrinkage which ultimately leads
to increase in intracellular viscosity.5 The apoptosis inducing
property of H2S is vital to study as it is also linked with antiꢀ
cancer properties of H2S.6 Thus, visualization and monitoring
of H2S induced apoptosis in cellular systems would be
important to understand the biological effects of H2S.
and acts as fluorescent molecular rotor. To the best of our
knowledge this is the first report where H2S induced apoptosis
phenomenon is investigated by a fluorescent probe. Moreover,
the designed probe
function i.e., detects H2S as well as monitors H2S induced
apoptosis. (ii) Probe can detect H2S upto 35 nM, which is one
1 has many advantages: (i) It performs dual
1
of the best among the reported probes. (iii) It can detect both
exogenous and endogenously generated H2S in C6 glial cells.
(iv) Probe
1 undergoes complete intracellular reduction with
exogenous and endogenous H2S which was proved by carrying
out mass spectral studies of cell fluid.
The synthesis of the probe
S5ꢀS6) and structure of the probe
spectroscopic data (ESI†, Fig. S1 to S8). The molecular
recognition behaviour of probe towards H2S was studied by
1
is given in scheme S1 (ESI†, Page
1
was confirmed by its
1
Recently, a number of fluorescent probes have been reported
for the detection of H2S but most of these probes suffer from
the limitations of poor detection limit and their biological
application is limited to exogenous detection of H2S only and
furthermore, in none of these reports H2S induced apoptosis has
been studied7. Based on these observations, we were then
interested to design a probe which performs dual function i.e.,
detects H2S as well as monitors H2S induced apoptosis in
cellular systems. For achieving this target, we envisaged that if
we could design a molecule that have molecular rotation
property and an effective H2S reaction centre then it will be
able to perform dual function. The molecular rotation property
is required to monitor H2S induced apoptosis as the rotation
gets restricted in viscous medium created during apoptosis.8
Keeping this in mind, we designed and synthesized a mesoꢀ
UVꢀvis and fluorescence spectroscopy in H2O: DMSO (8:2,
v/v) buffered with HEPES, pH = 7.02. The absorption spectrum
of probe
1 (5.0 ꢁM) exhibits two absorption bands at 495 nm
and 350 nm (ESI†, Fig. S9 and S10) corresponding to the S0ꢀS1
and S0ꢀS2 transitions with molar extinction coefficients of 4.65
×104 and 1.45×104 Mꢀ1 cmꢀ1 respectively.
H2S
0 µM
(A) (B)
H2S
60 µM
substituted bodipy derivative
1 which has a phenyl ring
Fig. 1: Fluorescence response of probe
(8:2, v/v) buffered with HEPES, pH=7.02; on addition of H2S (0ꢀ
60 ꢁM); λex= 470 nm and λem = 515 nm.
1
(5.0 µM) in H2O: DMSO
connected to bodipy core. Further, an azide group is introduced
in this derivative as it is known to undergo reduction in the
presence of H2S. We envisaged that the resulting reduced
This journal is © The Royal Society of Chemistry 2012
J. Name., 2012, 00, 1-3 | 1

ChemComm
Page 2 of 4
View Article Online
COMMUNICATION
Journal Name
DOI: 10.1039/C5CC02984H
Upon addition of H2S (Na2S was used as H2S source) to the fluorescence enhancement on lowering the pH from 7.4 to 4
solution of probe , the decrease in the absorption band at 495 (ESI†, Fig. S21a). On further lowering the pH from 4 to pH 2,
nm as a function of H2S concentration is observed (ESI†, Fig. there was slight enhancement in fluorescence intensity of probe
S11). The fluorescence spectrum of probe exhibits an (0.91 folds). To find out the reason why there was a
emission band at 515 nm when excited at 470 nm. Upon negligible fluorescence enhancement on lowering the pH from
1
1
2
addition of H2S (0ꢀ60 ꢁM) to the solution of probe
emission band at 515 nm decreases with the increase in the found to be 3.87 (ESI†, Fig. S21b). From this pK
1
the 7.4 to 4, we determined the pK
a
value of probe
2
a
which was
value and
concentration of H2S (Fig. 1). This decrease in fluorescence pH studies, it is clear that probe
emission with addition of H2S at 515 nm is attributed to (i) pH of the environment.11,13
2 is not much sensitive to the
photoꢀinduced electron transfer (PeT) from nitrogen atom to Further, we also studied time dependent response of the probe
1
photoꢀexcited bodipy moiety which leads to quenching of towards H2S. It was observed that 80% of fluorescence
fluorescence emission. (ii) The product formed after reduction quenching takes place within 20 min of addition of 60 ꢁM of
acts as fluorescent molecular rotor and the energy is dissipated H2S at a time (ESI†, Fig. S22). The time dependent
by nonꢀradiative pathways due to the free rotation of mesoꢀ fluorescence quenching indicates that probe
substituted phenyl ring (Scheme 1).
1
is an efficient
probe for monitoring the changes in H2S level in living
systems. The detection limit of probe for H2S was found to be
H2S
reactive
Change
in life
1
35 nM (ESI†, Fig. S23). Having done all this, we were then
interested to study the H2S induced apoptosis and for this, we
studied the fluorescence and fluorescence lifetime behaviour of
.
site
time on
changing
viscosity
probe
2 in different methanol: glycerol fractions and it was
Viscosity
sensitive
observed that the fluorescence intensity and decay time
increases with increase in viscosity of the medium which
proves that reduced product probe
S24 and S25). We then studied the fluorescence spectra of
probe in the presence of H2S (60 µM for 30 min) in different
methanol: glycerol fractions. significant increase in
fluorescence intensity was observed with increasing glycerol
fractions (ESI†, Fig. S26). However, in the absence of H2S no
significant enhancement in fluorescent intensity and
2 acts as FMR (ESI†, Fig.
Scheme 1: Designing strategy for monitoring H2S and H2S induced
apoptosis.
1
A
To investigate whether the probe
under physiological conditions, the absorption and fluorescence
response of the probe was also studied with reactive oxygen
1 can selectively detect H2S
1
species like ClOꢀ, H2O2, tBuO•, HO• and biothiols like cysteine,
glutathione and homocysteine but no significant change was
observed in the presence of these analytes in the UVꢀvisible and
fluorescence spectra (ESI†, Fig. S12, S13, S14 and S15). To
fluorescence lifetime of probe
1 was observed with increasing
glycerol fractions (ESI†, Fig. S27 and S28). These results
clearly indicate that the fluorescence enhancement is due to
reduced product
2 which may be used for monitoring H2S
test the practical applicability of the probe
1 we also carried out
induced apoptosis in biological systems.
the competitive experiments in the presence of H2S mixed with
other analytes but no noticeable change in the fluorescence
emission was observed in comparison with or without any other
Since, it has been reported that viscosity increases during
apoptosis in biological systems,5b,12 so we were interested to
study the H2S induced apoptosis in C6 glial cells. At first, we
analytes (ESI†, Fig. S16). Thus, probe
1 acts as an efficient
performed MTT assay which showed that probe
1 is nonꢀtoxic
fluorescent probe for the selective detection of H2S in mixed
in nature and may be safely used for biological studies (ESI†,
Fig S29). Initially, we carried out experiment for exogenous
detection of H2S and for this purpose; the cells were first
aqueous environment. Further, to confirm the reduction of
1
probe
probe
1
1
, we also carried out the mass and H NMR studies of
in the presence of H2S. The appearance of a peak at m/z
incubated with probe
with 20 µM H2S for 20 min. The quenching of fluorescence
emission was observed in cells, which was due to the reduction.
1 for 20 min at 37 ºC and then treated
416.21 (ESI†, Fig. S17) in the mass spectrum confirms the
formation of reduced product 2. The H NMR spectrum shows
an appearance of new peak at 3.7 ppm corresponding to amino
protons and upꢀfield shift of 0.1 and 0.4 ppm for the Ha and Hb
protons which confirms the conversion of azide group to amino
moiety in the presence of H2S (ESI†, Fig. S18 and S19). We
1
Overlay
Green Channel
DIC
(c)
(b)
(a)
also studied the e
ff
ect of pH on the recognition behavior of
S2ꢀ
towards H2S which exists in equilibrium with HSꢀ
/
(f)
(e)
(d)
probe
1
at pH 7. It was found that fluorescence quenching occurs at a
faster rate in basic pH than in acidic pH (ESI†, Fig. S20). This
fast quenching in fluorescence emission at pH > 7 is due to the
existence of H2S as HSꢀ under basic conditions which has more
reducing power in comparison to H2S which exists in acidic
pH.7e,10 Further, we also studied the effect of pH on the
Fig. 2: Cell images of C6 cell lines. Row 1 (a,b,c): cells were treated
with probe (1.0 ꢁM) for 20 min at 37 °C. Row 2 (d,e,f); cells were
treated with probe (1.0 ꢁM) for 20 min and then incubated with
H2S (20 ꢁM) for 20 min. Images were taken at λex =488 nm and λem
range 500ꢀ550 nm. All images are taken at 40X magnification.
1
1
fluorescence behaviour of probe
2. There was a negligible
2 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 2012

Page 3 of 4
Journal Name
ChemComm
View Article Online
COMMUNICATION
DOI: 10.1039/C5CC02984H
of azide group to amino moiety (Fig. 2) and (ESI†, Fig. S30).
2ꢁg/ml lipopolysaccharide (LPS) for 24 hrs. LPS stimulated
Further to monitor the H2S induced apoptosis; we increased the
cells were then incubated with probe 1 for 20 min and cell
incubation period and concentration of H2S in cells. When we imaging was performed. The cell imaging results showed an
increased the incubation period of H2S from 20 to 60 min, then enhancement in fluorescence emission instead of quenching
instead of quenching, an enhancement in fluorescence emission
(Fig. 5) and (ESI†, Fig. S33). The enhancement in fluorescence
intensity was observed (Fig. 3) and (ESI†, Fig. S30). From the intensity is due to the fact that LPS treatment stimulated cells to
above results, it may be concluded that when cells were generate H2S and this process continued for 24 hrs and during
incubated with exogenous H2S for longer period then apoptosis this time, the endogenously generated H2S initiates apoptosis in
gets initiated in cells and as a result of which, the intracellular cells as a result of which the viscosity of cells increases leading
viscosity gets increased leading to enhancement in fluorescence to enhancement in fluorescence emission of probe
emission of reduced product via restriction of rotation. confirm the reduction of azide in cells by LPS stimulation, we
Further, on increasing the concentration of H2S from 20 µM to carried out mass spectral studies of cells and observed a peak at
100 µM and incubation for 20 min, again fluorescence m/z 416.23 which proved that the reduced product is formed
enhancement was noticed in C6 cells (ESI†, Fig. S31) which by reduction of probe by endogenously generated H2S (Fig.
may be due to the fact that higher concentration of H2S ). Thus, from the above results, we propose that probe shows
2. Further, to
2
2
1
4
1
enhanced the rate of apoptosis. Thus, we observed an turn off response with H2S before apoptosis and turn on
enhancement in fluorescence intensity in both cases i.e., on response after initiation of apoptosis.
Overlay
Green Channel
DIC
longer incubation period of H2S and at higher concentration of
H2S. On the other hand, the other possibility of increase in
fluorescence emission in cells may be due to the binding of
(c)
(a)
(d)
(b)
azide moiety of probe
prevent azide to undergo reduction in the presence of H2S.
1 with intracellular proteins which will
(e)
(f)
Overlay
(c)
Green Channel
(b)
DIC
(a)
Fig. 5: Cell images of C6 cell lines. Row 1 (a,b,c): cells were treated
with probe (1.0 ꢁM) for 20 min at 37 °C. Row 2 (d,e,f); cells were
stimulated with LPS (2ꢁg/ml LPS) for 24 hrs at 37 °C and then
treatment with probe (1.0 ꢁM) 20 min. Images were taken at λex
1
(d)
(f)
(e)
1
=488 nm and λem range 500ꢀ550 nm.
Further, to confirm that the fluorescence turn on response is due
to H2S induced apoptosis, we performed nuclear condensation
studies by staining the cells with DAPI (4',6ꢀdiamidinoꢀ2ꢀ
(i)
(h)
(g)
Overlay
DAPI
Green Channel Zoomed
Fig. 3: Cell images of C6 cell lines. Row 1 (a,b,c): cells were treated
with probe (1.0 ꢁM) for 20 min at 37 °C. Row 2 (d,e,f): cells were
treated with probe (1.0 ꢁM) for 20 min and then incubated with
H2S (20 ꢁM) for 20 min. Row 3 (g,h,i): cells were treated with probe
(1.0 ꢁM) for 20 min and then incubated with H2S (20 ꢁM) for 60
1
1
1
min. Images were taken at λex =488 nm and λem range 500ꢀ550 nm.
To rule out this possibility, we carried out mass spectral studies
of cell cytosol. Cells were treated with probe
1 and incubated
with 20 µM of H2S for 30 min and then cells were lysed and
centrifuged to obtain the cell supernatant. The mass spectrum of
this supernatant solution showed
a
peak at 416.23
corresponding to reduced product
S32).
2 (Fig. 4) and (ESI†, Fig.
Cells + probe 1
Probe 1
Probe 1 + exogenous H2S Probe 1 + endogenous
416.2304
Exogenous
Reduced
product
Endogenous
H2S
416.2359
Reduced
product
417.2403
417.2325
Fig. 6: Cell images of C6 cell lines showing nuclear condensation by
H2S induced apoptosis. Cells were first treated with probe (1.0 ꢁM)
then incubated with H2S (20 ꢁM) for different time intervals of 10,
20, 60, 120 min. Images were taken at λex =488 nm (for probe) and
405 nm (for DAPI) with λem range 500ꢀ550 and 430ꢀ480 nm.
416
418
416
417
1
Fig 4: Intracellular mass analysis for confirmation of reduction of
probe with exogenous and endogenous sources of H2S in cells.
1
Thus, the mass spectral studies prove that probe undergoes
1
phenylindole), which selectively stains the nucleus and the
changes in nuclear morphology during apoptosis could be
observed by this stain12. The cells were first incubated with
reduction with H2S in cells and thus rule out the possibility of
binding of azide to proteins. Further, to detect endogenously
generated H2S in living cells, the cells were stimulated with
This journal is © The Royal Society of Chemistry 2012
J. Name., 2012, 00, 1-3 | 3

ChemComm
Page 4 of 4
View Article Online
COMMUNICATION
Journal Name
DOI: 10.1039/C5CC02984H
probe for 20 min and then treated with H2S for different time UPE/UGC/CSIR program for their fellowship. UGC, grants
1
intervals of 10, 20, 60 and 120 min. Finally, cells were washed under UPE and CPEPA schemes and DBT, GOI grant under
and then stained with DAPI, which selectively stains the SubꢀDIC are acknowledged for providing infrastructural
nucleus and depicts the changes in nucleus morphology facilities to carry out the research work.
associated with apoptosis. Cell imaging results showed that
upto 20 min, no significant change in nuclear morphology was
observed and the fluorescence intensity of cells decreased
which is due to the reduction of azide to amine. Further, on
increasing the incubation period to 60 and 120 min, the cells
showed prominent nuclear condensation as well as increase in
fluorescence intensity (Fig. 6). The intensity analysis also
showed that at 120 min maximum fluorescence intensity was
observed (ESI†, Fig. S34). The nuclear condensation proves
that cells underwent apoptosis. We also carried out
fluorescence lifetime imaging (FLIM) studies to prove the
phenomenon of apoptosis. FLIM studies were carried out in C6
Notes and references
aDepartment of Chemistry, UGC Sponsored Centre for Advanced
Studiesꢀ1, Guru Nanak Dev University, Amritsar, Punjab, India.
Eꢀmail: mksharmaa@yahoo.co.in
bDepartment of Biotechnology, Guru Nanak Dev University, Amritsar,
Punjab, India.
Electronic Supplementary Information (ESI) available: [Experimental
details, NMR, mass spectra, fluorescence spectra and time resolved
fluorescence spectra]. See DOI: 10.1039/c000000x/
1
(a) D.G. Searcy and S.H. Lee, J. Exp. Zool., 1998, 282, 310; (b) J.
Furne, A. Saeed and M. D. Levitt, Am. J. Physiol. Regul. Integr.
Comp. Physiol., 2008, 295, 1479.
2
(
a
) Y. Han, J. Qin, X. Chang, Z. Yang and J. Du, Cell. Mol.
Neurobiol., 2006, 26, 101; ( ) P. Kamoun, M.ꢀC. Belardinelli, A.
Chabli, K. Lallouchi and B. ChadefauxꢀVekemans, Am. J. Med.
Genet., 2003, 116A, 310; ( ) K. Eto, T. Asada, K. Arima, T.
Makifuchi and H. Kimura, Biochem. Biophys. Res. Commun., 2002,
293, 1485; ( ) Z. Lou, P. Li, K. Han, Methods in Molecular Biology.,
2015, 1208, 97ꢀ110.
) J.H. Zhang, Z. Dong and L. Chu, J. Periodont Res., 2010, 45, 71;
) T. Murata, K. Yaegaki, W. Qian, M. Herai, B. Calenic, T. Imai, T.
b
glial cells in the presence of probe
1 and then incubating the
c
cells with 20 µM of H2S for different time intervals of 10, 20,
60 and 120 min. We observed that upto 20 min, no significant
change in decay time was observed in comparison to control
(cells incubated with only probe 1). Further, on increasing the
incubation time to 60 and 120 min, the decay time was
d
3
(a
(b
Sato, T. Tanaka, T. Kamoda and H. Ii, J. Breath Res., 2008,
2,
increased dramatically (Fig. 7 and ESI†, Fig. S35). This result
017007; (c) G. Yang, X. Sun, and R. Wang FASEB J., 2004, 18
,
justifies that the reduced product
2 acts as FMR and shows an
1782.
10 min
20 min
0 min
4 (a
) I. Aoyama, B. Calenic, T. Imai, H. Ii and K. Yaegaki, J. Periodont
Res., 2012, 47, 365; (b) C. Kobayashi, K. Yaegaki, B. Calenic, N.
Ishkitiev, T. Imai, H. Ii, I. Aoyama, H. Kobayashi, Y. Izumi, and M.
Haapasalo, J. Endod., 2011, 37, 479.
5 (
a
) R. Baskar, L. Li, and P. K. Moore, FASEB J., 2007, 21, 247; (
Hortelano, M.L. GarcıꢀMartın, S.Cerda, A. Castrillo, A.M. Alvarez
and L. Bosca, Cell Death Differ., 2001, , 1022.
(a) L. Zhang, Q. Qi, J. Yang, D. Sun, C. Li, Y. Xue, Q. Jiang, Y.
b) S.
5
2
ns
ns
120 min
60 min
8
6
7
Tian, C. Xu, and R. Wang, Oxid. Med. Cell Longev.,2015, Article ID
636410, in press.
Fig. 7: Fluorescence life time imaging (FLIM) in C6 cell lines
showing change in life time of probe by H2S induced apoptosis.
Figure showing the changes in life time when the cells treated with
probe (1.0 ꢁM) then incubated with H2S (20 ꢁM) for different time
(a) B. Peng, C. Zhang, E. Marutani, A. Pacheco, W. Chen, F. Ichinose
1
and M. Xian, Org. Lett., 2015, 17, 1541; (b) F. Yu, P. Li, P. Song, B.
Wang, J. Zhao and K. Han, Chem. Commun., 2012,48, 2852–2854; (c)
P. Zhang, J. Li, B. Li, J. Xu, F. Zeng, J. Lv and S. Wu, Chem. Commun.,
2015, 51, 4414; (d) V. S. Lin, W. Chen, M. Xian and C. J. Chang,
Chem. Soc. Rev., 2015, DOI: 10.1039/C4CS00298A; (e) C. Liu, B. Peng,
S. Li, C. M. Park, A. R. Whorton and M. Xian, Org.Lett., 2012, 14,
2184; (f) S. I. Reja, N. Kumar, R. Sachdeva, V. Bhalla and M. Kumar,
RSC Adv., 2013, 3, 17770; (g) B. Wang, P. Li, F. Yu, J. Chen, Z. Qu and
K. Han, Chem.Commun., 2013,49, 5790; (h) Z. Lou, P. Li, and K. Han,
Acc.Chem.Res., 2015, 48, 1358−1368.
1
intervals of 10, 20, 60, 120 min. Images were taken at λex =488 nm
(for probe) with λem range 500ꢀ550 nm.
increase in decay time with increasing viscosity during
apoptosis13. Therefore, the reported probe
for detecting H2S as well as for monitoring H2S induced
apoptosis phenomenon in living cells.
In conclusion, we designed and synthesized a bodipy based
1 is an efficient tool
8
(
J
a
) H. Zhu, J. Fan, M. Li, J. Cao, J. Wang, and X. Peng, Chem. Eur.
., 2014, 20, 4691; ( ) Z. Yang, Y. He, J. H. Lee, N. Park, M. Suh,
W. S. Chae, J. Cao, X. Peng, H. Jung, C. Kang, and J. S. Kim;
J.Am.Chem.Soc., 2013, 135, 9181; ( X. Yin, Y. Li, Y. Zhu, X. Jing,
Y. Li and D. Zhu, Dalton Trans., 2010, 39, 9929.
) M. A. Haidekker and E. A Theodorakis, J. Biol.Eng., 2010,
) Y. Zhao, T. D. Biggs and M. Xian, Chem. Commun., 2014, 50
11788; ( ) O. Kabil and R. Banerjee, J.Biol.Chem., 2010, 285, 21903.
11 ( ) H. Yu, Y. Xiao, and L. Jin, J. Am. Chem. Soc., 2012, 134,17486.
12 ( ) J. A. Collins, C. A. Schandl, K. K. Young, J. Vesely, M. C.
Willingham,
13 (
2014,
dual functional fluorescent probe
H2S and can also monitor H2S induced apoptosis in cellular
systems. Further, the potential of the probe to explore H2S
induced apoptosis phenomenon was studied by FLIM studies
which showed that during apoptosis the decay time of probe
(amine) increases due to restriction of rotation in viscous
intracellular matrix. Hence, the designed probe reported
herein is an efficient tool for the detection of H2S and H2S
induced apoptosis in cellular system and will attract more
attention of the scientists to discover more sensors which can
detect H2S as well as monitor H2S induced apoptosis.
1 which selectively detects
b
1
c
)
2
9
(
a
a
4,11.
,
10 (
1
b
a
a
J
.
Histochem Cytochem., 1997 ,45, 7923.
) T. Liu, X. Liu, D.R. Spring, X. Qian, J. Cui and Z. Xu, Sci. Rep.,
, 5418.
a
4
MK & VB acknowledge the financial support from DST and
CSIR, New Delhi. NG, SIR and MG acknowledges
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 2012