NEWS & VIEWS doi:10.

1038/nature24755

C H E MICAL B IO LO GY background. Second, light scattering caused

by particulate matter generally decreases with

Organic dyes for longer wavelengths. A theoretical study3 has

predicted a 100- to 1,000-fold improvement in
image resolution at centimetre depths of tissue

deep bioimaging
when imaging in the SWIR range (1,350nm),
compared with imaging at a near-IR
wavelength (850 nm).
Several challenges must be overcome to
Small-molecule organic dyes that fluoresce in the short-wave infrared region of translate theory into experimental reality.
the spectrum could improve the resolution of in vivo bioimaging methods. Such One technological issue is that conventional
dyes have now been made by adapting those that fluoresce visible light. light detectors (such as the sensors used in
most digital cameras) lose effectiveness for
wavelengths beyond 1,000 nm. The use of
MARTIN J. SCHNERMANN absorb and emit light at wavelengths greater detectors that function in the SWIR range has
than 1,000nm. been restricted, in part by national-defence

F
luorescence-based imaging methods Red light is preferentially transmitted concerns, because they are used in night-
have transformed the way that scientists through tissue which is why your hand vision devices and in other military applica-
visualize and interpret biological events. lights up red when you hold a torch behind it. tions. However, sensitive cameras equipped
An enduring goal is to make fluorescence To take advantage of this property, there have with SWIR sensors based on the semiconduc-
imaging broadly useful in complex physi- been substantial efforts to develop probes and tor indium gallium arsenide are now sold for
ological settings, especially for clinical appli- in vivo imaging techniques that use far-red and biomedical use, and the availability and cost of
cations. One strategy is to use fluorescence near-IR wavelengths (6501,000nm). these cameras are likely to improve.
wavelengths between 1,000 and 2,000nano- However, imaging in the SWIR range The central chemical challenge in this field
metres the short-wave infrared (SWIR) offers important benefits for in vivo applica- is to identify probes that emit in the SWIR
region of the electromagnetic spectrum. But tions, compared with that in the far-red and range. A seminal study4 published in 2009
organic molecules that absorb and emit these near-IR, for two main reasons. First, SWIR revealed that surface-modified carbon
wavelengths are needed to realize the full produces less autofluorescence light emit- nanotubes absorb light in the near-IR range
potential of SWIR imaging1. Writing in Ange- ted naturally by biological structures that and emit at SWIR wavelengths. Imaging
wandte Chemie, Cosco et al.2 report the bright- have previously absorbed light so that experiments with these compounds revealed
est known organic small-molecule dyes that images are produced against a nearly black dramatic benefits in resolution compared with
conventional fluorescence imaging at near-IR
wavelengths greatly improving the reso-
a lution of through-skull imaging of the brain
vasculature in mice5, for example. And earlier
N N
this year, a class of quantum dots (fluorescent
+
n I nanometre-scale particles that contain heavy
Cyn; n = 3, 5 or 7 metals) was reported6 that enable a range of
in vivo applications for SWIR imaging. These
b Ph Ph Ph Ph two types of nanomaterial finally brought the
benefits of SWIR imaging to fruition.
+
O O O
+
Cl O
ClO4 ClO4 But clinical protocols for optical bioimaging
almost exclusively use fluorescent small
n molecules (known as fluorophores) as imag-
Me2N NMe2 Me2N NMe2 ing agents, rather than nanomaterials7. This
Flavn; n = 3 or 5 Flav7
is because fluorophores generally have low
toxicity, are cleared from the body by well-
c Cy7 Flav3
Cy3 Cy5 Flav5 Flav7 defined mechanisms, and can be straight
forwardly targeted to biomolecules. The same
400 600 800 1,000 1,200 1,400 1,600 1,800 2,000 nm is probably true for molecules that fluoresce
Visible Near-infrared Short-wave infrared in the SWIR region, but few such molecules
are known, and even fewer that have useful
properties for biological applications8.
Figure 1 | Increasing the wavelength of light emitted and absorbed by dyes. a,The Cy dye series of Making bright SWIR molecules is a
compounds is used for optical fluorescence imaging. The dyes consist of indolenine heterocycles (red)
challenging problem for organic chem-
connected by polymethine linkers (orange). I, iodide ion. b,Cosco et al.2 modified the structures of these
ists. Solving it requires advanced synthetic
dyes, replacing the indolenines with dimethyl-flavylium heterocycles (blue), and dubbing the resulting
methods combined with design insights from
compounds Flav dyes. Me, methyl group; Ph, phenyl group; ClO4, perchlorate ion. c,Each Flav dye emits
and absorbs light at a longer wavelength than the analogous Cy dye. Notably, Flav7 emits and absorbs light theoretical chemistry. Enter Cosco and col-
in the short-wave infrared region (SWIR) of the electromagnetic spectrum. The authors used Flav7 as a leagues, whose strategy was to make rational
probe for SWIR fluorescence bioimaging, which offers higher resolution and visualization of deep tissue modifications to the molecular scaffold of
than can be achieved using visible-light fluorescence imaging. Wavelengths are shown in nanometres. fluorescent dyes already used for optical
(Adapted from Fig. 1 of ref. 2.) imaging using visible and near-IR wavelengths.

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 RESEARCH NEWS & VIEWS

Perhaps best known as the Cy dye series9,
these compounds contain two hetero
cycles (rings that contain atoms other than
carbon) connected by 3, 5 or 7 methine units
(=CH groups; Fig.1a). They are exception-
ally good light absorbers, a key property that
contributes to these molecules being excel-
lent emitters in response to light irradiation.
Cosco and co-workers gave Cy dyes a chemical
makeover by replacing the conventionally used
heterocycles (indolenines) with dimethyl-
flavylium (Flav) heterocycles (Fig. 1b),
devising a concise synthetic route by which to
prepare the new compounds.
The resulting dyes have several potentially
useful properties for bioimaging applica-
tions. The wavelengths of light absorbed and
emitted by the new dyes are shifted roughly
200nm towards longer wavelengths that is,
towards the infrared region compared with
the corresponding Cy dyes (Fig. 1c). Notably,
the compound that contains 7 methine units
in its linker emits light in the 1,100-nm range,
and is the brightest small-molecule SWIR dye
reported so far. The authors used a formulation
of this dye to visualize deep vasculature in
a mouse.
A major goal for clinical optical imaging
is to enable surgeons to readily visualize, and
then remove, tumour tissue10. The improved
tissue penetration and optical resolution of
SWIR imaging should help surgeons to iden-
tify metastases secondary tumours formed
at sites away from the primary tumour that
would be hard to find using existing methods.
Additional advances are required on multiple
fronts before SWIR imaging can be broadly
used for clinical applications. For example,
fluorophores that have high solubility in water
must be identified this might be possible
by making modifications to the molecules
reported by Cosco and colleagues. Other
key objectives include finding ways to target
fluorophores to specific biomolecules, and
developing fluorescent probes that can be
switched on in response to biological stimuli.
These chemical advances will have to be paired
with emerging developments from engineer-
ing and biomedical research. For example, a
portable, ideally hand-held, SWIR imaging
apparatus is needed that can be readily used
in a surgical setting. Nevertheless, Cosco and
colleagues study provides a key breakthrough
required to move SWIR imaging towards
general clinical use.
Martin J. Schnermann is in the Chemical
Biology Laboratory, National Cancer Institute,
National Institutes of Health, Frederick,
Maryland 20850, USA.
e-mail: martin.schnermann@nih.gov
1. Thimsen, E., Sadtler, B. & Berezin, M. Y.
Nanophotonics 6, 10431054 (2017).
2. Cosco, E. D. et al. Angew. Chem. Int. Edn 56,
1312613129 (2017).
3. Lim, Y. T. et al. Mol. Imaging 2, 5064 (2003).
4. Welsher, K. et al. Nature Nanotechnol. 4, 773780
(2009).
5. Hong, G. S. et al. Nature Photonics 8, 723730 (2014).
6. Bruns, O. T. et al. Nature Biomed. Eng. 1, 0056 (2017).
7. Lavis, L. D. & Raines, R. T. ACS Chem. Biol. 9,
855866 (2014).
8. Antaris, A. L. et al. Nature Mater. 15, 235242
(2016).
9. Gorka, A. P., Nani, R. R. & Schnermann, M. J.
Org. Biomol. Chem. 13, 75847598 (2015).
10. Nguyen, Q. T. & Tsien, R. Y. Nature Rev. Cancer 13,
653662 (2013).

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