Laboratory of Nanobiology
Institute for Protein Research

Professor

Yoshie HARADAyharada

URL: http://www.protein.osaka-u.ac.jp/nanobiology/

Our body is made up of many cells.
The proper functioning of each cellallows us to live in good health.
Knowing the state inside a cell andknowing what is happening inside acell is very important forunderstanding life. Therefore, weare developing methods to measure
the environment of the nano region
inside the cell using small
fluorescent diamond particles,
measuring the local temperatureinside the cell, and investigating how
the temperature change affects
the cell function. We also provide afluorescence microscope that can
image protein secretion fromindividual cells in real time, and are
conducting collaborative researchwith multiple laboratories.

Quantum biosensing using
fluorescent nanodiamond
It is necessary to develop a nano-sized
sensor to investigate how nanometricenvironments (e.g. temperature, magneticand electric fields) affect the biochemical
reactions inside single cells. Fluorescent
nanodiamonds (FNDs) containing nitrogenvacancy centers (NVCs) are emerging as anovel type of fluorescent probe due to thestable fluorescence showing no photobleaching (Fig. 2). Besides, their uniquequantum properties lender them as apromising nanometric biological sensor.
Our group has developed an ODMR
microscope enabling the optical read-out of
the quantum states of FNDs. Using theODMR microscope, we have demonstrated
temperature and thermal conductivity
measurements of living single cells. We
currently investigate the mechanism how
cells sense temperature by using chemicallycontrolled FND composites.

Developing techniques for intracellulartemperature measurement
The temperature is a fundamental parameterin biology and affects metabolism andbiorhythm. However, temperature fluctuationsin space and time in individual cells has been
largely unknown. We develop methods toquantitatively image the temperature using thefluorescence microscopy and various types offluorescent “nanothermometers” that are
based on fluorescent polymers (Fig. 1),fluorescent nanodiamonds, fluorescent
nanoparticles or fluorescent molecules. In thisresearch topic, we are studying the biologicalrelevance of the thermal fluctuation. We also
aim to apply our methods to issues inbiomedical studies.

Figure 2. Illustrationof a fluorescent
nanodiamond and anitrogen-vacancy
centerAnalysis of immune responses using
live-cell imaging of secretion activity
Intercellular messengers such as hormones,
cytokines, and extracellular vesicles play animportant role in the information exchange
between cells and the collaborative regulationof the biological systems that organize humanbody. We have developed "live-cell imaging ofsecretion activity (LCI-S)" to visualize the true
state of cell secretion by combining
fluorescence sandwich immunostaining andtotal internal reflection fluorescencemicroscopy (TIRFM) techniques (Fig. 3).
Since LCI-S is suitable for primary cultured
mouse cells, human clinical biopsied cells, EScells, iPS cells, etc., it is expected to be usedfor various applications such as functional
hematology in precision medicine, phenotypic
screening and toxicity evaluation in drug
discovery, quality control of cell therapy andevaluation of efficient control of differentiationin regenerative medicine. We are also working
on a social implementation of our technology(Live Cell Diagnosis, Inc.).
Looking inside the cells that make up ourLooking
insidetheprotein
cells thatmake upareourbody,variousmolecules
working
there.
In orderto knowbody,
variousproteinmolecules
are aboutworking
there.
In order it
toisknow
organisms,
itorganisms,
very about
importanttois very
importantto protein
investigate
how protein
investigate
howmolecules
work.
So far,work.
we havedevelopeda developed
method formolecules
So far,we havea methodfordirectly observing
howproteindirectlyobserving
how protein
molecules
molecules
workausing
a microscope,
andwork using
microscope,and haveinvestigatedhowproteinsworkwell.
haveinvestigatedhowproteinsworkwell.
Recently,
we arealsoalsoobserving
cellscells
in inRecently,
we areobserving
order
to understandthe thecellscells
whereorderto understandwhereproteinswork.
Littleby little,
we want
to toproteinswork.
Littleby little,
we want
approachwondersof life.
approach
the thewondersof life.

Institute for Protein Research, Osaka University
3-2 Yamadaoka, Suita, Osaka 565-0871, Japan
TEL: +81-6-6879-8627
FAX : +81-6-6879-8629

Figure 1.

Figure 3. Schema of LCI-S: live-cell imagingof secretion activity

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