e. gracilis gciy-egfp with high - rigidity chip€¦ · scm250 (iii) patterning of etching mask....
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𝑑𝑑
0
20
40
60
80
0 50 100 150 200
Dis
plac
emen
t 𝑑𝑑[µ
m]
Time [µs]
11050
100
𝜏𝜏: µsµsµsµs
Response speed :< 16 µs
Evaluation of response speed
0
40
80
120
160
200
0 20 40 60 80 100 120Sorti
ng w
idth
𝑤𝑤[µ
m]
Rising time 𝜏𝜏 [µs]
1009080
∆𝑉𝑉𝑚𝑚𝑚𝑚𝑚𝑚: VVV
Sorting width :> 100 µm
Evaluation of sorting width
Vertical sheath inletHorizontal sheath
inletSample inlet
Fixation hole
Interestport
Primingport
AttachmentGlass membrane
Piezoelectric actuatorOn-chip dual membrane pump
On-chip cell sorting by high-speed local-flow controlusing dual membrane pumps
Shinya Sakuma, Yusuke Kasai, Takeshi Hayakawa, Fumihito AraiNagoya University Micro-Nano Mechanical Science and Engineering
Corresponding author: Shinya SakumaE-mail: sakuma@mech.nagoya-u.ac.jp,URL: http://www.biorobotics.mech.nagoya-u.ac.jp/〒464-8601 Furo-cho, Chikusa-ku, Nagoya-shi, Aichi-ken, JapanMicro-Nano Mechanical Science and Engineering, Arai lab.TEL: +81-52-789-5220, FAX : +81-52-789-5027
Acknowledgement:This study was supported by a Grant-in-Aid from the ImpulsingParadigm Change through Disruptive Technologies Program (ImPACT).References:[1] S. Sakuma et al., On-chip cell sorting by high-speed local-flow control using dual membrane pumps, Lab on a Chip, 2017, DOI:10.1039/c7lc00536a[2] Nitta et al., Intelligent Image-Activated Cell Sorting, Cell (2018), https://doi.org/10.1016/j.cell.2018.08.028
On - chip dual membrane pumpswith high - rigidity chip
High-rigidity chip
e.g.) CTC(Circulating Tumor Cell)
http://bradyurology.blogspot.jp/2014/05/circulating-tumor-cells-for-prostate.html
e.g.) Euglena
http://www.goda.chem.s.utokyo.ac.jp/research_japanese.html
Rare cells in massive cells
Highly specific cellsin a group
・Biofuel・Nutritional food・Cosmetic product
・Diagnosis・Prognosis・Drag development
http://www.jst.go.jp/impact/serendipity/concept.html
http://www.jst.go.jp/impact/serendipity/concept.html
Background : Cell sortingcell sorting
cultivation
cell sorting
Size of target cell/particle/droplet [µm]
Equi
vale
nt th
roug
hput
[Hz]
100 0 50 100 150
101
102
103
104
105
Y. Chen, Small, 2014
S. H. Cho, Lab Chip, 2010
Z. Cao, Lab Chip, 2013T. D. Perroud, Anal. Chem., 2008
R. W. Applegate Jr., Lab Chip, 2006
L. Ren, Lab Chip, 2015
L. Rao, Sens. Act. B, 2015
Y. Chen, Analyst, 2013
Z. Cao, Lab chip, 2013
S. Li, Anal. Chem., 2013
S. W. Lim, PLOS ONE , 2015
E. gracilisGCIY-EGFPA. Sciambi, Lab Chip, 2015
Main-flow Local-flow
Local-flow
Pump A
Pump B
Pump A
Pump B
Pump A
Pump B
Local-flow
Local-flow
Pump A Pump A Pump BPump BPump A Pump B
Sorting principleNon-sorting
stateSorting state
t = 0 μs
t = 80 μs
t = 40 μs
t = 120 μs
t = 200 μs
t = 160 μs
100 μm
t = 0 μs
t = 40 μs
t = 80 μs
t = 120 μs
t = 160 μs
t = 200 μs
(v) Patterning of etching mask for sand blasting
(vi) Sandblasting
Glass
SU-8Si
OFPRCr Au
SCM250
(iii) Patterning of etching maskafter anodic bonding
(i) Patterning of etching mask
(ii) HF etching and cleaning
(iv) DRIE
(vii) Anodic bonding
Packaging
Base/Cover layer Cover layer
Channel layer
Fabrication process
Sorting experimentsTarget: E. gracilis Target: GCIY-EGFP
𝜏𝜏 = 100 µs𝑑𝑑 = 33 µm
𝑤𝑤 𝑤𝑤
𝜏𝜏 = 1 µsEvaluation of flow control
Borosilicate glass
SiliconBorosilicate glass
31 kHz
High-speedcamera
Chip Stage
Laser(488 nm)
Photomultiplier(PMT)
Half Mirror
Halogen lamp
White light
Mirror
Polarizer
Dichroicmirror
Notch filter
High-voltageamplifier
Detectioncircuit
PZT1 PZT2
Analyzer
Wasteport
Sorting system
スライド番号 1
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