ウィスパリング ・ギャラリー・モードによるリガンド―受容体相互作用の新規高感度検出法
High performance novel sensing of receptor-ligand interactions by whispering gallery
mode resonators

代表研究者

Cevayir COBAN

IFReC, Osaka University

協同研究者

Sahin Kaya OZDEMIR

Washington University in St. LouisLan YANG

Washington University in St. Louis

Malaria is one of the most important diseases in the world causing high levels of morbidity and mortality, killing
almost 1 million people every year. Development of rapid and sensitive diagnosis techniques for malaria is still an
important need for robust and effective treatment regimens. Hemozoin is a bio-crystalline metabolite of Plasmodiumparasites. During blood circulation of Plasmodium parasites, hemozoin is continuously produced, released and captured
by immune cells, activating immune-mediated signaling cascades. Because hemozoin crystals are the unique signatureof malaria parasites, detection of hemozoin in circulating blood or serum with high sensitive platforms, even at thesingle nanocrystal/molecule resolution, is the ideal platform for the early diagnosis of malaria disease. Furthermore, thehighly sensitive platforms which can detect hemozin accurately would be beneficial to study hemozoin’s interaction
with immune cells.
Whispering-gallery-mode (WGM) resonators have recently been emerged as new platforms for sensing DNA,
proteins and pathogens such as viruses. With an international collaboration, we have recently explored the use of WGM
resonators both in air and liquid as label-free detection and measurement of hemozoin crystals with single crystal
resolution. We have further explored the detection of changes during hemozoin and various molecule interactions.

intensity via the confinement of light field) of optical

研究目的
It is extremely important to develop highly sensitive

resonances.

As

technologies to detect very small molecules, both in air

high-fidelity

opticaland water, for various applications such as early

phenomena with superior sensitivity (1, 2). WGM

pathogen detection which can prevent outbreaks. Malaria

resonators have been shown to be feasible platform foris one of the important diseases in the world causing highthelevels of morbidity and mortality. Development of rapid

single-particle resolution (3, 4), as well as nucleic acidand sensitive techniques for malaria diagnosis is still aninteractions.

important need for robust and effective treatment

detectionsuch,of

WGM

resonators

measurements

viruses

andofenablebiological

nanoparticles

atHemozoin is a nano-sized crystalline by-product ofPlasmodium parasites. During blood circulation ofregimens.
Whispering-gallery-mode (WGM) resonators have

Plasmodium

parasites,

hemozoiniscontinuously

versatile

produced, released and captured by immune cells.

nano/micro-photonics sensing platform that relies on theBecause hemozoin crystals are the unique signature ofultrahigh-quality (Q) (enabling longer interaction time)malaria parasites, detection of hemozoin in circulating

and the microscale mode volume (enabling high optical

blood or serum with high sensitive platforms, even at theemerged

asanewandvery

single nano-crystal/molecule resolution, will enable theearly diagnosis of malaria disease (5). Hence, we haverecently used WGM resonators for the first time inbiological fluids todetect hemozoin crystals,

aby-product of Plasmodium parasites, for the early
detection of malaria disease (5).
Hemozoin is a biologically active molecule thatinteracts

withvarious

immunecells

andimmune-mediated signaling cascades. In this research
project, we have investigated the possibility of using
high-Q WGM resonators (which can detect hemozin
accurately) to study hemozoin’s interaction with2) It has recently been shown that hemozoin crystals play aimmune system molecules.

role in host immune system modulation by malaria
parasites and interact with the host immune system. PI

研究経過

Coban and the others have tried to elucidate these1) The mode splitting in active and passive WGM opticalinteractions;however,resonators as a novel self-referencing technique forlimitations.

For

detecting perturbations in close proximity to themicroscopy to investigate hemozoin’s effect on a single

resonatorbycell level (6). The Raman spectroscopy as a label-free

co-investigators Ozdemir and Lang. Using the mode

method detected the biochemical changes occurring insplitting technique, detection and single-shot size

macrophages during the first few hours of hemozoin

measurement of individual nano-scale objects, including

uptake. There is a high possibility that innate arm of thepolystyrene, potassium chloride and gold particles as

immunesystemwhichiswell as Influenza A virions, with the minimum detectable

immunologically

activecomponents

size of 20 nm has been achieved (4). This technique has

microbes are activated in these macrophages after the

been applicable in aquatic environment that hemozoin

hemozoin uptake. One question that remained to be

crystals could be detected easily with WGM microtoroid

addressed is: How can the receptor-mediated recognition

resonators even in biological fluids (5).

of hemozoin cooperate at the level of ligand recognition?

has

been

introduced

recently

thereinstance,

have

we

been

haveevolved

technical

usedtoRaman

recognize

derivedfromTo perform WGM sensing for the detection of

A previous study by our group showed by using circularhemozoin crystals (Figure), microtoroids were fabricateddichroism (CD) spectrum analysis that there are somefrom silica-on-silicon wafer with diameters D=~50 μm

interactions between TLR9 protein and hemozoin crystals

and hemozoin crystals deposited on a microtoroid(7). This implies light-polarization dependent response ofresonator. The mode splitting experiment was performed

these crystals. This is interesting, because we have shownin an aquatic environment. The results of this study

in our studies with WGM resonators that the response ofhighlighted that highly sensitive early-phase detection of

the WGM microresonator is very much dependent on themalaria infection from serum is possible with WGM

orientation of the hemozoin in the sensing volume and theresonators.

polarization of the probing light field. Therefore, thisimplies that sophisticated methods such as mode splittingin active and passive WGM optical resonators can befurther utilized as a novel self-referencing technique fordetecting perturbations due to the presence of hemozoin
crystals in close proximity to the resonator to probe suchinteractions at single crystal level. Currently, we aredesigning

newfluidic

platform

toemployouralready-established WGM platform for studying suchProcNatlAcad

interactions.

16;111(37):E3836-44.

SciUSA,

2014,

Sep

3. Zhu J, Özdemir SK, He L, Chen DR, Yang L. Single考察
WGM resonators are very sensitive microscalevirusandnanoparticle

size

spectrometry

bywhispering-gallery-mode microcavities. Opt Express,

photonic systems that can enable high-performance

2011, 19(17):16195–16206.

sensing platform to detect biological materials and their

4. He L, Ozdemir SK, Zhu J, Kim W, Yang L. Detecting

interactions. Ultra-high-Q and small mode volumes

single viruses and nanoparticles using whispering gallery

provide a superior sensitivity surpassing the performancemicrolasers. Nat Nanotechnol, 2011, 6(7):428–432.

of traditional interferometric or Surface Plasmon

5. Kim W, Ozdemir SK, Zhu J, Faraz M, Coban C, Yang

Resonance (SPR) techniques. Interestingly, plasmonic

L. Detection and size measurement of individualcomponents can be easily incorporated to WGM

hemozoin nanocrystals in aquatic environment using aresonators to make use of the best of two platforms:

whispering gallery mode resonator. Opt Express, 2012,

Highly localized field of plasmons (nanoscale mode

20(28):29426–29446.

volume in plasmonic particles systems versus microscale

6. Hobro AJ, Pavillon N, Fujita K, Ozkan M, Coban C,

mode volume in WGM resonators) and ultra-high-Q

Smith NI. Label-free Raman imaging of the macrophage(over 10 million in WGM versus a few thousands inresponse to the malaria pigment hemozoin. Analyst, 2015,

plasmons). Now, these WGM resonators are used for

Mar 16; 140(7):2350-9.

wide range of applications including detection of7. Coban C, Igari Y, Yagi M, Reimer T, Koyama S, Aoshi

biohazardous microbes in air or label-free sensing ofT, Ohata K, Tsukui T, Takeshita F, Sakurai K, Ikegami T,

single molecules not only in air but in biological fluids

Nakagawa A, Horii T, Nuñez G, Ishii KJ, Akira S.

such as hemozoin detection in aqueous environment. We

Immunogenicity of Whole Parasite Vaccines Against P.

have also shown the performance enhancement in WGM

falciparum Involves Malarial Hemozoin and Host TLR9.

resonators using intrinsic gain mechanisms such as

Cell Host Microbe, 2010, 7(1):50-61.

Raman gain in the material from which WGM resonators

8. Peng B, Özdemir ŞK, Liertzer M, Chen W, Kramer J,

are fabricated to compensate optical losses to increase

Yılmaz H, Wiersig J, Rotter S, Yang L. Chiral modes andsensing resolution as well as to turn WGM resonators

directional lasing at exceptional points. Proc Natl Acad

into microlasers to significantly enhance detection andSci U S A. 2016 Jun 21;113(25):6845-50.

simplify the detection system for nanoparticle detection(8). We hope that this innovative tool will help

研究の発表

understanding how microbes are recognized and further

口頭発表

designing molecules for the development of new drugs1.

Coban C: Malarial hemozoin in infection andand/or vaccines at least against malaria. We are nowvaccination. Invited talk at the 10th Meeting of thedeveloping our systems to detect hemozoin interactions

Japanese Vaccine Adjuvant Research Consortium,in real time by monitoring the response of WGM

Senri Life Science Center, Osaka, JAPAN, January

miroresonators and microlasers.

24th, 2017.
2.

Ozdemir SK: Whispering-gallery-mode resonators

参考文献

and their applications for nanoscale sensing and1. Vahala KJ. Optical microcavities. Nature, 2003,

measurement. Invited talk at the SPIE Photonics
West, Integrated Photonics and Resonators I, San

424(6950):839–846.

Francisco, CA, USA, January 28-February 2,

2. Özdemir ŞK, Zhu J, Yang X, Peng B, Yilmaz H, He L,Monifi F, Huang SH, Long GL, Yang L. Highly sensitive
detection of nanoparticles with a self-referenced andself-heterodyned whispering-gallery Raman microlaser.

2017.
3.

Coban C: Tissue-specific Immunopathology during
Malaria. Invited talk at Protein Island Matsuyama

(PIM)

InternationalSymposium

2016.

University, Matsuyama, JAPAN, September 16-17,
4.

5.

9-11, 2016.

Ehime
9.

Coban C: Tissue-specific Immunopathology during

2016.

Malaria. Invited talk at the 5th NIF Winter School on

Coban C: Tissue-specific Immunopathology during

Advanced Immunology, Awaji Island, Hyogo,Malaria. Invited talk at the John Curtin School ofJAPAN, January 17-22, 2016.

Medical Research (JCSMR), Australian National

10. Coban C: Lessons from Malaria Immunology: A

University (ANU), Canberra, AUSTRALIA, AugustTale of Hemozoin. Invited talk at Preston M. Green19, 2016.

Department of Electrical & Systems Engineering,Ozdemir SK: Innovative photonic technologies fromWashington University in St. Louis, St. Louis, MO,sensing to fundamental physics using whispering

USA, August 6, 2015.

gallery resonators. Invited talk at St. Louis
6.

7.

University, St. Louis, MO, USA, May 26, 2016.

誌上発表

Coban C: Tissue-specific Immunopathology during

1.

Malaria. Invited talk at Molecular Immunology &against Plasmodium vivax infection via LAP-like

Immunogenetics Congress (MIMIC-III), Antalya,degradation of sporozoites. Proc Natl Acad Sci U S A,TURKEY, April 27- 30, 2016.

2016, Jun 10. pii: 201607007.

Ozdemir SK: Frequency Splitting in Whispering

2.

Lee MSJ, Igari Y, Tsukui T, Ishii KJ, Coban C.

Gallery Microlasers for High Performance Sensing.

Current status of synthetic hemozoin adjuvant: A

Invited talk at the Materials Research Society (MRS)

preliminary safety evaluation. Vaccine, 2016, Apr

Spring Meeting, Phoenix, AZ, USA, March 28th-19;34(18):2055-61.

April 1st, 2016.
8.

Lelliott PM, Coban C. IFN-γ protects hepatocytes

3.

Lee MSJ, Maruyama K, Fujita Y, Konishi A, Lelliott

Coban C: Immunopathology of malaria: search for

PM, Itagaki S, Horii T, Lin JW, Khan SM, Kuroda E,

new drug targets. Invited talk at the 89 Annual

Akira S, Ishii KJ, Coban C. Plasmodium products

Meeting of the Japanese Pharmacological Society,persist in the bone marrow and promote chronic

Pacifico Yokohama, Yokohama, JAPAN, Marchbone loss. Science Immunology, 2017, in press.

th