Transcript Document

細胞晶片研究發展的國際
合作經驗
Prof. Ching-Hsing Luo (羅錦興)
Department of Electrical Engineering
National Cheng Kung University
Tainan, Taiwan
1 - Virtuoscope & Wireless Mixed Signal Biochip Lab
From Tissue To Single Cells
• Tissue is a group of cells, cannot see single
one easily.
• Basic researches of DNA and virus move on
the investigation of single cells.
• Nanotechnologies can see inside the single
cells and operate DNA easily.
• Computing power enables the simulation
easily in comparison to experiments.
2 - Virtuoscope & Wireless Mixed Signal Biochip Lab
International Collaboration
Necessary Conditions
• Have the niche, replaceable not easily.
• Have a teamwork in Taiwan to collaborate
with.
• Find the mutual needs in the project.
• Not just learn their techniques.
• Open the mind if you can.
3 - Virtuoscope & Wireless Mixed Signal Biochip Lab
My Niche: Single Heart Cell Model
Luo-Rudy model in 1991 &1994 Circ. Res.
3 papers with SCI cited 333, 410, 186 in total and 184, 280, 88 in recent five years
4 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Action Potential by Model Simulation
Action potentials for K o=7, 5.4, 4, and 3 mM
Membrane Potential (mV)
50
0
-50
[K]o=7 5.4
-100
0
100
200
300
time(msec.)
4
400
3 mM
500
600
1. A standard procedure of model simulation has been developed to
evaluate the accuracy of simulation results
2. A physiological simulation fake is discussed.
5 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Drug screening Platform- 1st project
Invited by Prof. Miqin Zhang, UW
by Miqin Zhang
High performance of integrated cell-based sensors for drug screening
with impedance measurement only.
6 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Mission
• Medical Researches
Stem cell, Neuron cell, Cancer cell for Drug
screening, Differentiation or cultivation control,
Gene therapy
• Engineering Technologies
Chip – micro/nano (channel and pumping)
Sensor and actuator – micro/nano scale
Cell Modeling – Ischemia, drug effects
7 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Team Up
Domestic:
1. Cell-based microchip platform –李清庭、羅
錦興、魏憲鴻、 鍾宜璋
2. Cell Model -羅錦興、吳勝男、鄧君豪
3. Bio-application – 宋瑞珍副校長、林茂村院
長、吳勝男
International: Univ. of Washington
• Miqin Zhang: drug screening
• Meldrum: HIV, AIDS applications
8 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Commercial Microarray chip
9 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Single Cell Chip Platform
Microfluid Flow
Single Cell
Microelectrode
Micromixer
Microchannel
Microelectrode
Microvalve
Microvalve
Micropump
Flow sensor
Micropump
1. Automatic Protocol control (concentration
clamp)
2. Voltage clamp for ionic channel measurement
10 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Passive Mixing in Microfluidic Channel
by Prof. Wei
q
Channel Width
Setup
Width 300 mm
Open angle q=60
11 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Mixing Results-1
12 - Virtuoscope & Wireless Mixed Signal Biochip Lab
α
R
q
Mixing Results-2
13 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Cell Clamping Microchip Platform
Silicon oxide
Differentiation or cultivation protocols
Electrode
Sucking pressure
Silicon
Glass
14 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Cell Attachment and Rejection
H2O/OH-
thiourea
Br
Electrode
n OH
n OH
HS
reflux
I2
Cell
BrCH2CH2OH
POCl3
O P Cl
(III)
HO
n S
Cl
OPO
n S
Br
S
(IV)
NH3
O
OH
NH2
(VI)
S
n OH
O
OH
O
S
(II)
O
n S
EtOH
O
Br
Channel
OPO
(I)
n OPO
HO
Br
O
OPO
n OPO
OH
HO
+
NH2 H3C N CH3
CH3 Br
n=9
(CH3)3N
O
n S
(V)
S
n OPO
HO +
H3C N CH3
CH3
Br
Cell Rejection Compound
15 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Cell-Attachment Technique
by Nano-Imprinting.
by Prof. Chung and Prof. Luo
•
Using flat PDMS imprint SH-(CH2)11COOH on Au
cell favoring coating
Imprinting：
PDMS
Time≒ 60sec
Chemical bond:
SH-(CH2)11COOH
Au
16 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Attachment Structure
(a)Before adhering
cell
flowing
Platform
silicon
(b) After adhering
cell
platform
Au on silicon base
17 - Virtuoscope & Wireless Mixed Signal Biochip Lab
1.6 μm
Electrode Shape Effect
• Cell name : human CD34+ Progenitor Cell provided
by Prof. Lin, Chi-Mei Medical center.
• Cell is stretched to platform corner
18x18 μm 2
18 μm diameter
square platform
circle platform
18x36 μm 2
ellipse platform
18 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Budget Consideration
• Usually not a big funding, just initiate the
collaboration with let-go rule.
• 應用於藥物篩選的細胞微晶片平台和細胞模
型 NSC 94-2614-B-006 -037, $1,200,000 per
year for six professors.
• Work together and come up something for
future budget application in both sides.
• International collaboration experiences are
important to open your territories and mind.
19 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Nano-tag studies on LDL atherogenic
effects on VEC- 2nd project
• Applying Nano National-type Project, passed
the first phase with good order.
• Initiated by Prof. Chen, Baylor College of
Medicine, Huston, USA.
• Team up by Prof. Tzeng, Director of Nano
center, NCKU.
• Hosted by Prof. Chen, Vice-Dean of NTU
hospital.
20 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Aim-1: Determine the receptors for L5
on EC membrane
• L5 or LDL5 is invented
by Prof. Chen’s team at
Baylor College of
Medicine.
• Prof. Chen’s team
provides L5 to NTU and
NCKU team for singlecell and clinical studies.
PAFR and LOX-1 may be activated by
L5 signals sequentially
21 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Aim-2: Temporal tracing of L5
localization in EC
• Nano-tag on L5 by Prof.
Yeh, NCKU
• High-resolution Optical
Microscope by Prof.
Tzeng, NCKU Nano
center.
Left panel: aggregates of L5-nano tagged
Right panel: a single L5 molecules-nano tagged
22 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Aim-3: Determine calcium-signaling
effects during L5 apoptotic process
• Build up VEC model
to find the ionic
activities during L5
apoptotic process.
• Create a transparent
single-cell platform
for L5-tagged VEC
studies.
L5
?
?
?
?
KATP
BKCa
KCa
NSC
TRPC
NCX
SKCa
LoX-1
IRK
[Ca2+]i
ER
?
?
PAFR
?
IKCa+
GPCR
SOC
SERCA
SERCA
Cl-
Mitochondria
CFTR
VRAC
BCl
CaCC
RACC
CNG
Na+
Ω
Caveolar
SERC
A
Na+/Ca2+
M
Ca2+ mechanical sensor
(GPCR,
VAMP-2)
An EC model with ion channels and organelles.
23 - Virtuoscope & Wireless Mixed Signal Biochip Lab
60 GHz 無線生醫感測網路晶片系統
-3rd project
• International Collaboration by CRC (Canada),
CIC, NTU, NTHU, NCTU, NCKU.
• 60 GHz GaAs experiences from CRC (Canada)
exchanged with 60 GHz CMOS experiences
from universities and CIC in Taiwan.
• Invited talk at IEEE conference about this
project.
• Leaded by 呂學士教授, EE, NTU, honored by
solicited invitation talk in 2006 ISSCC.
24 - Virtuoscope & Wireless Mixed Signal Biochip Lab
60 GHz Antenna Design and
Temperature RFIC
• 60 GHz antenna design in LTCC and
CMOS
• Near-body communication RFIC design
• In-body communication RFIC design.
• Physiological Signal RFIC design for
portable devices.
25 - Virtuoscope & Wireless Mixed Signal Biochip Lab
Good luck to All of You!
and
Thanks for Your Attention
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