ラン藻のオンチップ機械的特徴量計測

ラン藻のオンチップ機械的特徴量計測
○学長谷川貴之1
正佐久間臣耶1
非七谷圭2
非魚住信之2
正新井史人1
1．名古屋大学大学院工学研究科 2. 東北大学大学院工学研究科
Concept
Background & Purpose
ラン藻 (Cyanobacteria)
Kdp
Ktr
Synechocystis sp. PCC 6803
K+
MscL
 Fixation of CO2
 Alterative energy
AqpZ
K+
(Mechanosensitive Cannel) H2O
Cell membrane
Deformation：
On-chip probe
K. Nanatani et al., J. bacterial, 2014
• Floating cell
• Diameter: around 2 µm
• Need to apply nN force to deform
5 mm
Na+
NhaS3
Conventionally…
Group cell analysis
 Driven by external actuator
 Output force: > 1 kN
 Resolution: nm order
Example of group cell analysis
Force sensing :
On-chip sensor
Purpose Measuring mechanical characteristics of a single cyanobacterium
Fabrication of Robochip
Trap & Transportation :  3 DOF non-contact manipulation
 Positioning accuracy: nm order
Optical tweezers
(a) Patterning of etching mask
(b) Etching of borosilicate glass by RIE,
and removal of etching mask
(e) Bonding borosilicate glass and device layer by anodic bonding
(c) Patterning of etching mask
(f) Patterning of etching mask
(d) Etching of device layer by DRIE, and removal of etching mask
(g) Etching of handle layer and BOx
layer, and removal of etching mask
Side view
World‐to‐chip Sensor beams
interface
Piezoelectric Actuator
SOI wafer
Handle layer 225 µm
BOx layer
1 µm
Device layer 7.5 µm
100 µm
Glass
Transportation of a single cyanobacterium
Mechanical characterization
Holographic Optical Tweezers System
Lens
Lens
Polarizer
Yb Fiber Laser:
1064 nm
0 sec
1 sec
Iris
Beam Expander
2 sec
5 sec
100
Force measured by force sensor
4
Force [nN]
Onda et al., Opt. Express, 2012
SLM
Dichroic
Mirror
Displacement of pushing probe
5
Spatial Filter
Displacement [μm]
Objective Lens
3
2
1
60
40
20
0
0
0
5 μm
80
100
200
300
0
10
20
30
40
50
Deformation ratio [%]
Time [sec]
Pushing
probe
Hertzian contact theory
Sensor
beam
P
Transportation
150 sec
0 sec
Normalized specimens
Positioning

R
300 sec
Young’s modulus
On‐chip
probe
On‐chip
sensor
EC 
3(1   2 ) P
2
4 R1 /・
( / 2 ) 3 / 2
Young’s modulus [kPa]
Trapped cell
BG11 medium
70
60
50
40
30
20
10
0
BG11 + 1.5 M Sorbitol
1.5
1.6
1.7
1.8
1.9
Diameter [μm]
Mechanical characteristics were measured!!
Moiré fringe
本研究に関するお問い合わせ先：長谷川貴之（Takayuki Hasegawa)
E‐mail: [email protected]‐u.ac.jp, URL: http://www.biorobotics.mech.nagoya‐u.ac.jp/
名古屋大学大学院工学研究科マイクロ・ナノシステム工学専攻新井研究室
TEL: 052‐789‐5026, FAX : 052‐789‐5027
Acknowledgements：
本研究は科学研究費（24686032）の
支援をうけて行われたものです．
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