SURGEON AT WORK

Intraoperative Fluorescent Cholangiography

Using Indocyanine Green: A Biliary Road Map
for Safe Surgery
Takeaki Ishizawa, MD, Sumihito Tamura, MD, Koichi Masuda, MD, Taku Aoki, MD, PhD,
Kiyoshi Hasegawa, MD, PhD, Hiroshi Imamura, MD, PhD, Yoshifumi Beck, MD, PhD,
Norihiro Kokudo, MD, PhD

Unlike blood vessels, the biliary tract lies in the Glissonian
sheath and is buried in the perivascular connective tissue,
so it is difficult to clearly visualize and isolate it during
hepatobiliary surgery. Intraoperative cholangiography
(IOC), which was originally introduced by Mirizzi1 in
1937, has been widely used to delineate the biliary tract
anatomy in this setting. For example, routine IOC was
recently recommended during cholecystectomy to prevent
bile duct injury.2-5 IOC is also considered an essential pro-
cedure during donor hepatectomy because it enables the
bile duct to be divided at the appropriate level to ensure
wider and fewer residual orifices.6-10 But conventional ra-
diographic IOC is disadvantageous in that it exposes the
patient and the medical staff to radiation and usually re-
quires a large and expensive C-arm fluoroscopy machine
and the additional human resources involved.11
Recently, intraoperative angiography using a fluorescent
imaging technique with IV injection of indocyanine green
(ICG) has been used to assess coronary artery bypass graft
patency.12-15 This technique is based on the principle that
ICG binds to plasma proteins and that protein-bound ICG
emits light with a peak wavelength of about 830 nm when
illuminated with near-infrared light.16,17 Because human
bile also contains plasma proteins that bind with ICG,18 we
hypothesized that fluorescent images of the biliary tract
could be obtained with intrabiliary injection of ICG. We
also hypothesized that IV injection of ICG would provide
fluorescent images of the biliary tract without necessitating
Disclosure Information: Nothing to disclose.
Supported by grants from the Ministry of Education, Culture, Sports, Science
and Technology of Japan (Grant No. 18790955 and No. 17591377), the
Scientific Research from the ministry of Health, Labour, and Welfare of Japan
(Grant No. 18230201), and the Japanese Society for Advancement of Surgical
Techniques.
Received July 26, 2008; Revised September 23, 2008; Accepted September
24, 2008.
From the Hepato-Biliary-Pancreatic Surgery Division, Department of Sur-
gery, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
Correspondence address: Norihiro Kokudo, MD, PhD, Hepato-Biliary-
Pancreatic Surgery Division, Department of Surgery, Graduate School of
MedicineUniversity of Tokyo, 7–3–1 Hongo, Bunkyo-ku, Tokyo 113-8655,
Japan.
catheterization of the bile duct because ICG is excreted
exclusively by the liver, and biliary excretion of ICG con-
tinues from several minutes to as long as 20 hours after IV
injection.19 Here we describe novel fluorescent IOC tech-
niques with intrabiliary or IV injection of ICG for safer
hepatobiliary operations.
METHODS
Patients
Subjects included 13 patients who underwent donor right
hepatectomy (n ⫽ 4) or liver resection for hepatobiliary
malignancy requiring IOC to divide the hilar bile ducts
(right hepatectomy, n ⫽ 3; right lateral sectoriectomy,
n ⫽ 2; central bisectriectomy, n ⫽ 1; and partial hepatec-
tomy, n ⫽ 3), and 10 patients who underwent open cho-
lecystectomy for acute cholecystitis (n ⫽ 2), chronic cho-
lecystitis with gallstones (n ⫽ 6), or gallbladder carcinoma
(n ⫽ 2), at Tokyo University Hospital.
Administration of indocyanine green
In the 13 hepatectomy patients, ICG (0.025 mg/mL; Di-
agnogreen; Daiichi Sankyo Co) was administered into the
bile duct through a transcystic tube before division of the
hilar bile ducts. Before injecting the ICG, a small amount
of bile (1 mL or less) was aspirated into a syringe to pro-
mote binding between the bile proteins and ICG.
In the 10 cholecystectomy patients, 1 mL of ICG (2.5
mg/mL) was injected IV 1 hour before the operation
(n ⫽ 7) or at the time of conversion from laparoscopic to
open cholecystectomy (n ⫽ 3) to use the ICG excreted in
the bile as the source of fluorescence.
Fluorescent imaging techniques
The fluorescent imaging system (PDE; Hamamatsu Photon-
ics Co) is composed of a small control unit (322 ⫻ 283 ⫻ 55
mm; 2.8 kg) and a camera unit (80 ⫻ 181 ⫻ 80 mm; 0.5
kg). The camera unit comprises a charge-coupled device
camera that filters out light with a wavelength of less than
820 nm, and 36 light-emitting diodes with a wavelength of
760 nm. The camera imaging head was positioned 20 cm

© 2008 by the American College of Surgeons ISSN 1072-7515/09/$36.00

Published by Elsevier Inc. e1 doi:10.1016/j.jamcollsurg.2008.09.024

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e2 Ishizawa et al Intraoperative Fluorescent Cholangiography J Am Coll Surg

Table 1. Detectabilities of Intraoperative Fluorescent and

Abbreviations
Radiographic Intraoperative Cholangiography for Biliary Tracts
Fluorescent Radiographic
IOC ⫽ intraoperative cholangiography
IOC IOC
ICG ⫽ indocyanine green
Site of biliary tract n % n %
Intrabiliary injection of ICG
(13 hepatectomy
above the bile duct (Fig. 1) and the surgical lights were patients)
turned off (ceiling lights were kept on). At this level of CHD 13/13 100 13/13 100
brightness, the operative field was visible to the naked eye Confluence of the right
and on the television monitor. Fluorescent images of the and left hepatic duct 13/13 100 13/13 100
biliary tract were displayed on the television monitor. Preoperative intravenous
injection of ICG (10
After fluorescent cholangiography, radiographic IOC cholecystectomy
was performed using a C-arm fluoroscopy machine with patients)
intrabiliary injection of contrast material (Omnipaque) CHD 10/10 100 8/10 80*
and its detect abilities for the biliary tracts were compared Cystic duct 9/10 90
with those of fluorescent IOC. Right lateral sector branch
draining into the CHD† 4/4 100 4/4 100
RESULTS *Radiographic intraoperative cholangiography was unsuccessful because of a
failure to insert a transcystic tube in 2 patients with acute cholecystitis.
Fluorescent IOC showed the common hepatic duct in all †
This type of right lateral sector branch was observed in 4 patients.
the patients, regardless of the injection route of ICG (Table 1). CHD, common hepatic duct; ICG, indocyanine green; IOC, intraoperative
cholangiography.
Fluorescent IOC using intrabiliary injection of ICG delin-
eated the confluence between the right and left hepatic
ducts in all hepatectomy patients. The segmental branches and Supplementary Video 3) except for one with acute
of the intrahepatic bile duct draining into the right or left cholecystitis, in whom Calot’s triangle was not exposed
hepatic duct were also identified on the fluorescent images because of severe adhesion between the duodenum and the
in four patients; this information is helpful for appropriate hepatoduodenal ligament (Table 1). In another patient
division of the hepatic duct in donor hepatectomy (Fig. 2A with acute cholecystitis, fluorescent IOC identified the cys-
and Supplementary Video 1) and in partial hepatectomy tic duct and the common hepatic duct (Fig. 3B), although
for hepatocellular carcinoma (Fig. 2B and Supplementary radiographic IOC was unsuccessful because of a failure to
Video 2). insert a transcystic tube. The intervals between the injec-
Fluorescent IOC after preoperative IV injection of ICG tion of ICG and the initial examination ranged from 45 to
demonstrated the cystic duct before the dissection of 180 minutes (median 60 minutes). Fluorescence of the
Calot’s triangle in all cholecystectomy patients (Fig. 3A common hepatic duct lasted until closure of the abdomen
(90 to 310 minutes after the ICG injection). No adverse
reactions to the ICG were encountered.

Figure 1. Photograph of the operative field during fluorescent intra-
operative cholangiography.
DISCUSSION
Fluorescent IOC after intrabiliary injection of ICG showed
the confluence between the right and left hepatic ducts in
all hepatectomy patients. In addition, the procedure con-
ducted after preoperative IV injection of ICG enabled
identification of the cystic duct and the common hepatic
duct from before the dissection of Calot’s triangle to the
closure of the abdomen in cholecystectomy.
Our fluorescent IOC technique has several advantages
over the conventional radiographic examination. First, this
technique allows visualization of the biliary tract on the
images along with the surrounding structures in real time,
which helps surgeons to select the optimal point to transect
the cystic duct or hepatic duct. Second, fluorescent IOC
with IV injection of ICG enables identification of the cystic

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Vol. 208, No. 1, January 2009
Figure 2. Fluorescent intraoperative cholangiography after intrabili-
ary injection of indocyanine green (left) and intraoperative anatomic
view (right) during donor hepatectomy and repeated liver resection
for hepatocellular carcinoma. (A) Fluorescent intraoperative cholan-
giography demonstrated not only the confluence between the right
and left hepatic ducts, but also a tributary of the Couinaud’s seg-
ment V hepatic duct (B5) draining into the root of the right hepatic
duct (see Video 1). (B) Fluorescent intraoperative cholangiography
clearly delineated a branch of the segment IV hepatic duct (arrow in
schema), which was subsequently ligated and divided. It also delin-
eated the segment III and IV hepatic ducts (B3 and B4, respectively)
to be preserved and the right and left hepatic ducts (RHD and LHD,
respectively). Note that hepatocellular carcinoma itself showed flu-
orescence even before the start of the cholangiographic imaging,
probably because the indocyanine green that had been injected
intravenously for routine liver function tests a day before operation
was retained in the tumor (see Video 2).
duct without the necessity of dissection of Calot’s triangle
or insertion of a transcystic tube for contrast material in-
jection, a procedure that, by itself, can cause bile duct in-
jury.20 After IV injection of ICG, surgeons can obtain flu-
orescent images of the biliary tract only by placing the
camera imaging head over the biliary tract during cholecys-
tectomy. Third, the technique is very safe. It does not entail
exposure to radiation, and the reported risk of adverse re-
actions to IV injection of ICG is quite small (approxi-
mately 0.003% at doses exceeding 0.5 mg/kg).21 Lastly, it is
a simple and convenient procedure to perform. The sur-
geon does not require assistance to perform fluorescent
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Ishizawa et al Intraoperative Fluorescent Cholangiography e3
Figure 3. Fluorescent intraoperative cholangiography after preoper-
ative IV injection of indocyanine green (left) and the intraoperative
anatomic view (right) during cholecystectomy. (A) Fluorescent intra-
operative cholangiography demonstrated the cystic duct (CyD in
schema), the common hepatic duct (CHD) and the gallbladder (GB).
The cystic artery (CyA) and a lymph node (LN) could also be identified
as fluorescence defects (see Video 3). (B) Fluorescent intraopera-
tive cholangiography identified the cystic duct and the common
hepatic duct during cholecystectomy for acute cholecystitis.
IOC, and no space-occupying C-arm fluoroscopic ma-
chines are required.
Intraoperative ultrasonography is another alternative to
radiographic IOC that also enables less invasive and real-
time imaging of the biliary tract.3 But it requires much skill
to scan the biliary tract and to interpret B mode ultrasono-
graphic images.22 In addition, its ability to identify small
biliary strictures is limited.22,23 So fluorescent IOC has po-
tential advantages over ultrasonography in that it delineates
the biliary tract anatomy corresponding to its intraopera-
tive anatomic view without requiring special skills.
The major limitation of the fluorescent IOC is that it is
impossible to visualize deep-lying intrahepatic bile ducts or
extrahepatic bile ducts covered with surrounding organs
with this technique because of the limited tissue penetra-
tion of near-infrared light emitted by the current imaging
system. The ability to detect small bile duct stones with this
e4 Ishizawa et al Intraoperative Fluorescent Cholangiography
technique, which is another requirement for IOC, is also
questionable. But recent advances in imaging modalities,
such as magnetic resonance cholangiography and CT
cholangiography, have also made it possible to preopera-
tively delineate the anatomy of the biliary tract24 and to
detect stones in the bile duct.25 So we believe that fluores-
cent IOC can fulfill the objectives of IOC in the majority
of hepatobiliary operations. Even when conventional IOC
or ultrasonography is needed, the complementary use of
the fluorescent technique helps surgeons understand the
relationships between the biliary tract and other organs.
We consider that fluorescent IOC using ICG is a safe
and valuable procedure that provides a road map of the
biliary tract anatomy in real time for safe hepatobiliary
surgery. If the instruments are further refined and applied
to laparoscopes in the future, this unique IOC technique
may also be useful in laparoscopic cholecystectomy.
Author Contributions
Study conception and design: Ishizawa, Kokudo
Acquisition of data: Masuda, Aoki, Hasegawa, Imamura, Beck
Analysis and interpretation of data: Ishizawa, Hasegawa
Drafting of manuscript: Ishizawa, Tamura
Critical revision: Kokudo
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