細胞晶片研究發展的國際 合作經驗

1 - Virtuoscope & Wireless Mixed Signal Biochip Lab

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

2 - 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.

3 - Virtuoscope & Wireless Mixed Signal Biochip Lab

International CollaborationNecessary 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.

4 - Virtuoscope & Wireless Mixed Signal Biochip Lab

My Niche: Single Heart Cell ModelLuo-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

5 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Action Potential by Model Simulation

0 100 200 300 400 500 600-100

-50

0

50

Mem

bra

ne P

ote

ntial (m

V)

time(msec.)

Action potentials for Ko=7, 5.4, 4, and 3 mM

[K]o=7 5.4 4 3 mM

1. A standard procedure of model simulation has been developed to evaluate the accuracy of simulation results

2. A physiological simulation fake is discussed.

6 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Drug screening Platform- 1st projectInvited by Prof. Miqin Zhang, UW

by Miqin Zhang

High performance of integrated cell-based sensors for drug screeningwith impedance measurement only.

7 - 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

8 - 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

9 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Commercial Microarray chip

10 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Single Cell Chip Platform

Microfluid Flow

Micromixer

Micropump

Micropump

Single Cell

Microchannel

Microelectrode

Microelectrode

Microvalve

Microvalve

Flow sensor

1. Automatic Protocol control (concentration clamp)

2. Voltage clamp for ionic channel measurement

11 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Passive Mixing in Microfluidic Channel by Prof. Wei

Setup

Width 300 mOpen angle =60

Channel Width

12 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Mixing Results-1

13 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Mixing Results-2

R

α

14 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Cell Clamping Microchip PlatformDifferentiation or cultivation protocolsSilicon oxide

Electrode

Sucking pressure

Glass

Silicon

15 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Cell Attachment and Rejection

CellElectrode

Channel

Br OHn

OHn SH

I2 EtOH

OH Sn

S OHn BrCH2CH2OH

POCl3Br O P

O

Cl

Cl

O Sn

S On P

O

OP

O

O

BrOH

BrOH

O Sn

S On P

O

OP

O

O

N+ OH

N+OH

CH3CH3

CH3 CH3CH3

CH3Br

Br

O Sn

S On P

O

OP

O

O

NH2

OHNH2

OH

NH3

reflux

thiourea H2O/OH-

(I)

(II)

(IV)

n=9

(V)

(CH3)3N

(VI)

(III)

Cell Rejection Compound

16 - 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

PDMS

Time≒ 60sec

SH-(CH2)11COOH

Au

Imprinting：

Chemical bond:

17 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Attachment Structure

flowingcell

cell

platform

Au on silicon base

(a)Before adhering

(b) After adhering

1.6 μm Platform

silicon

18 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Electrode Shape Effect

18x18 μm 2

square platform

• Cell name : human CD34+ Progenitor Cell provided by Prof. Lin, Chi-Mei Medical center.

• Cell is stretched to platform corner

18 μm diameter

circle platform18x36 μm 2 ellipse platform

19 - 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.

20 - 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.

21 - 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 single-cell and clinical studies. PAFR and LOX-1 may be activated by

L5 signals sequentially

22 - 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 taggedRight panel: a single L5 molecules-nano tagged

23 - 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.

SERCA

?

SERCA

TRPCNSC SOC

NCX

M

Ca2+ mechanical sensor

LoX-1

ER

Mitochondria

L5

?

[Ca2+]i

PAFR

Na+/Ca2+

?

? ? ?

VRACBCl

CaCC

RACC

KATP

BKCa

SKCa

KCa

IRK

Cl-

CNG

GPCR

CFTR

SERCAIKCa

+

?

Na+

Ω

Caveolar

(GPCR,

VAMP-2)

An EC model with ion channels and organelles.

24 - 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.

25 - 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.

26 - Virtuoscope & Wireless Mixed Signal Biochip Lab

Good luck to All of You! and

Thanks for Your Attention