Well I'm going to be giving is very
complimentary to what Dr given.

Prior to my presentation where I'm
going to focus on development of

Microsystems and micro technologies that

allow you to interface with biological
cells in some way and the cells can come

in the form of a blood sample can come in
the form of tissue or dissociated cells.

We also are going to talk about as well.

These are systems that allow you to work
with again dissociated neurons as well as

she was so there's a lot of motivation for

microanalysis systems and you seen some of
these motivations from previous speakers

were interested in doing
this in the first place.

Now the primary systems with relationship

to this are that they
provide the opportunity for

the developing systems that are higher
performing than what we have today.

You know systems that may give us some
more precise analysis of cells and

what we can we can get with
conventional technologies or.

Functionalities together
to get enabling total

analysis system to do
a particular type of analysis.

So I really have the advantages of
having you know the small size so

the sample sizes can be small the amount
of reagents are small there are portable

they have the potential for
being disposable as well I

think the biggest advantage is
the integration of multiple functions

into these systems to create very
complex systems and along our lines so

this kind of gives you an idea of what
one of those systems might look like and

this is also system but
cellular sample preparation is part.

The system they started with the blood
sample coming in and we know that

the D.N.A. that's contained in the blood
is within the white blood cells and

other cells not the red blood cells so
part of this operation is

a self-serving operation is part
of the sample preparation part

of this total analysis system so again
this shows multifunction is going on.

Starting with a blood sample coming in.

Separating out clicked in the white blood
cells Gneisenau Y. blood cells collecting

the D.N.A. from those cells and
then going through processes like P.C.R.

to amplify the amount of D.N.A. and
then finally doing some type of a D.N.A.

separation maybe a letter from
the next step and the system so

it's a multi functional system and
so you have to be able to not only

introduce samples in regions you have
to be able to very precisely manipulate

the samples in the region so there's
mixers there's valves and pumps and

these systems along with the individual
functional compartments to give you the.

So in order to make those systems.

This is going to background to give you
an appreciation of the other components

are going to look at you have a couple of
different ways of making these systems

there's a hybrid approach is that's
what this is demonstrating where

the individual functions
are made on different chips and

then they're combined
together in a final system.

So these final systems have to have
the ability to create or to interface and

logically as well as fluid equally
between these individual from functional

compartments so
on this previous example you can imagine.

Maybe having one chip and

the phase extraction component for
extracting the D.N.A. in another chair and

so you could have kind of a platform
upon which you build these and

you can kind of mix and match the
functionalities on the generic platform.

Approaches to make these systems in a
monolithic fashion it's kind of any great

system in which times
the different functions are in

different layers of a multi
layer system so these are again.

Functional systems they
have bio fluids and

reagents and
various gases flowing through them.

They also have the integration of valve
ing operations around the county inner

layers of the system so it's just another
way of making this multi-functional and

I'm now also system.

It's a monolithic approach
compared to the hybrid approach.

So we're going to do today is we're
going to talk about some aspects of it.

Take a long time to talk about all the
aspects of the systems that we're going to

really function focus on is
the cell separation and and

collection component of this D.N.A.

analysis system this total
analysis system to make

the systems we don't want to forget the
packaging that's involved so there's and

while there's functional analysis
compartments down on these chips.

There's also interfaces that are required
in order to be able to get the fluids to

them to get the detectors
of the application

mechanisms to the P.C.
are chamber of the system.

So it's not only the chips but

it's also the interface this seems
rather mundane on the surface but

it's actually you know it's crippling if
you can't do this you know if you can

produce some way of interfacing between
the macro world and the micro world but

it's also requires a lot of
technology development to get there.

So this this is an example of kind
of a hybrid approach to the D.N.A.

analysis system which has a blood sample
coming in and ultimately going through

a number of steps and doing a letter for
a second out of the D.N.A.

has one of the functions on the second
chip the compartments are going to

talk about this compartment
to start with and

that this compartment does is
it takes a blood sample in and

it collects cells from the blood for
further analysis.

So this chip can actually
while it's being.

Uses a blood sample analyzer it can be
used for sampling of many different types

of suspended cell samples and examples or

swab samples and
different fluids from the body

separates out red blood cells and
this case disposes of them so

they go to a waste chamber and
they collect C Right blood cells and

other where nucleated cells
that may be in a blood sample

further analyzes those cells so
we can actually take those cells and

we can categorize them into different
types of my blood cells into different

types of rare blood cells as we'll
see later in this presentation so

purification of our sample and this gives
us further kind of the kind of visitation

and sorting of the system it's called
opinion spectroscopy it's really that

technique to get us down to the and I say
we're doing individual cell analysis.

So the first part of the system is going
to talk about the cell separator and

it's based on magnetic
properties of cells.

So the cells can obtain their magnetic
properties either naturally as is the case

with the red blood cells and invasion
attaching nano particles to these cells so

they could be perhaps cancer cells and
have a particular receptor that we know

that we want to bind and
then a particle to.

And then in a particle being
a magnetic nanoparticle

we then have a centrally a magnetic cell
but through this attachment process.

So Robert example that will show today is
with blood because blood has iron and iron

renders the cell magnetic doesn't

necessarily have to be red blood cells and
that we're using this magnetic force on.

So in this case what we're doing is
we're applying a magnetic field across

a microchip.

I can envision a very small micro channel
and in the center of that microchannel

there's a small very small wire magnetic
wire that we have to find and I see that

if you mount this represents that wire
that's in the center of the channel on

the ground that wire we have blood flowing
down the length of the channel and

apply a magnet or if we put
a magnet close to that flute Tanno

you know create a magnetic field that
actually has a great associated with that

close proximity to the wire which
is in the center of the channel so.

The gradient produces a magnetic
force is what it amounts to.

So the magnetic force is not generated
just from the magnetic field it's really

the gradient of the magnetic field
that produces a force in the case

of cells

away from the wire depending on whether
there are white blood cells already.

And actually and depending on the bow
that we operate the system as well

as of operation we can actually make

the red blood cells either be attracted to
the wire in the center of the channel or

be repelled away depending on
how we orient our magnets and

with respect to the system so
they call the diamagnetic and

the pair of magnetic capture for
the cells.

And so this is how the system
works near the brain

magnets from the side blood flowing
down the potato Here's the wire that we

describe in the channel in this case
we're operating on a pair of magnetic red

blood cells are opera are attracted to the
center and they end up flowing out this

center out of the I think Alan end
of the system right blood cells and

other nuclei three as has a base
example of the technology.

So this is about the system looks
like it's essentially the size of

a glass slide.

The system is built on a glass slide and
that's another advantage of it.

It's very portable there's
no wires connected to it.

It's simply requires
that you have a magnet

in close proximity to the separation
chair so this is input.

He goes down the length of
the separation channel and

this is the outlet that we see blown
up in this figure to the right.

So this is a magnet that used to
generate the magnetic field and

that's all it really takes so it's very
portable from that perspective there

is a macro scale version of the system
they require very high power

electromagnets top rate and
it isn't QUITE is

a separation efficiency is seventy percent
where at ninety nine percent Microsystems.

So this means separations look like
a first order separation system so

this is with a single wire running down
the length of a Micro Channel having

three outlets a thing in and

looking at seeing what happens with the
red blood cells in the printer magnetic

That means that the red blood cells
are being attracted to the center wire.

So this is without
a magnetic force applied.

It's pretty dumb distribution of
cells throughout reality channels and

once you've turned on the magnetic field
you start attracting the red blood cells

to the center channel and you see that
most of the red blood cells travel out

the Senate channel not all of them travel
out in this and this first order design.

I can show that just as some of the with
this project are only some of my

videos work and
they work in a cryptic way sometimes.

So in this case he shows a red blood
cells flowing down the center but

they're also raising a lot in the outer
one and out with three rows we really

would like to go on the two so
this is the wire design.

So what we have is we went
back to the drawing board and

we decided what we need to do.

We gave my presentation.

We go.

Try to do in order to make
the design work more efficiently and

one thing that we observed what we
were working with the first design is

that you know depending on
how fast we flew the sample.

We get a different separation
French officials say and

you wouldn't expect that because it
really relates to the residence time

of these cells within the magnetic force
a little more time it has to move toward

the wire the more you going to collect
in the stand the chance to channel.

So as you increase the speed you
decrease efficiency of the operation and

that's what you would expect.

So in this first water system
the operation efficiency was somewhere in

the upper sixty's as far as amount
of red blood cells that we collected

we did some of the experiments for
right blood cells as well.

Looking at collecting are we.

First of all we flew instantly
tag these white blood cells and

looked at how many were
able to collect in the

spirit are always going to be a pastor.

How many we can collect
in the center channel

one slide past.

Coming up with I can ask a design
mechanism so it looks very similar to

the first design but instead of having
a single wire down the center of

the chain or we actually have this
configuration of magnetic material and

that's in the center channel that
additional separation efficiency

out of the out of the system so
in general this is.

And saying it still has
an input channel through out

the red blood cells in the center outlet.

And what you can find is the experimental

results that the separation for instance
is much greater for this system then it is

for the single wire type system that
almost all of the red blood cells it's

ninety nine percent plus percent of the
blood cells travel down the center channel

and the white blood cells another
nucleus cells are forced down the outer

channels so using this what we call
the cascade design approach we're able to

get much higher separation fence and so
yes it was around ninety nine percent.

We've actually got a little bit better
than was shown in the state and

now it's around ninety nine percent
on the average separation fifteen C.

for removing blood cells and
these are not tagged in any manner simply

operating off of the native magnetic
properties of the red blood cells.

And so we did the same thing.

Looking at the separation fence and
see for

why blood cells I think this particular
video word is I was checking out

beforehand and you can see you
know why blood cells are one and

out and three of the system and red blood
cells are being collected then and so

the separation efficiency was
about ninety nine percent for

the white blood cells and
other nuclear cells in the outer channels.

And so something else we were
interested in looking at was whether or

not we could preferentially separate
cells that may be located in the blood.

You know externally aided cancer
cells are really where we did

our experiments we took
the case of breast cancer.

And we spiked the blood sample with
breast cancer cells and we did so for us.

So there we could look at the separation
efficiency with this system and

we found that about the same results with
this is to mess with with my blood cells

that are forced down Channel one and
Channel three

where they can then before the process
are collected for downstream analysis.

So it's good not only for
separating right.

But also collecting other rare
cells from a blood sample.

So what we do with those cells that we
correct is the second part of the system

that we had shown that chip and some
of the early slides it's the impedance

spectroscopy part of the star
the system and what we do

these analysis cavities.

So we have analysis cavities that
are made in a right format and

there's thousands of these and
individual way and

we have ways of down into these positions
and shows a vacuum port or a vacuum

port is used to pull the cell in position
we have other mechanisms as well but

in the end what happens is the cells are
located between two opposing electrodes so

they have conductors on
either side of the cell.

And we analyze the cells
impedance characteristics.

So the characteristics give us some
information about the cell itself and

really want to give this information
about the cell membrane and

the SO
what's going on within the cell as well.

And so this is kind of the electric

model through our cell position
between two electrodes on either side.

And if you study like cancer
biology you find that and

the case of cancer being
the progress of cancer.

What happens and many cells as
a become thinker they express.

They're more or less a particular
proteins on their surface and

so they undergo fairly
significant changes and

those changes are picked up by these
logical properties of the cell.

This is one of those systems as it's
just showing what it looks like

this is a silicon wafer it has an array of
these announcers Cabazon it closer this is

kind of a has for electrodes configured
and it shows no closer here and

this is a vacuum of all the poles down the
cell at the bottom of the analysis cavity.

And it's just a view of the packaging
is more engineering than anything.

I'll ask for.

But we did various tests
with the systems to look and

see to see how sensitive this technique
was and this is actually a very

difficult experiment because what we were
using was the same type of cell all these

experiments were done in the same type of
cell they were crowned with and cells.

And what we are doing is we are blocking
the potassium channels at different

different batches of these cells and
then analyzing their impedance properties.

So what we are looking for is changes
in the SO characteristics simply

based on changes in the channel activity
and ion channel activity within the cell.

And we're able to pick that up you can
see in this in this particular what's

called the phase part of the data we
can pick up a difference in what these

just based on channel activity.

It's a very difficult task because it's
not much changing there the amount

of channels that are on
the surface is only a very

small percentage of the total
surface area of the cells.

Are interesting.

I think from a standpoint
of disease detection.

Whereas the application
of this to breast cancer.

What we do is we look.

Breast cancer cells from going

through better static cells and looking
at the impedance properties of the cells

that have been collected
from the blood samples.

And this is what we found using the same
approach spectroscopy data we have

magnitude in phase we find that it's very
easy to pick up the difference in stages

in this case breast cancer using
this approach going from so.

Through static cells the N.D.A.

And before thirty five is and as you can
see there is significant differences in

the signature of the impedance of that
we get for these the cell to cell types.

It's because of all the changes
that are that are occurring within

the cell coming at the cell membrane
as well as what's Korean side the cell.

It's kind of a cumulative
effect this is so

measurement it's not it's not
really down to the molecular level

stage in the later stage breast cancer.

You're right they do
have those differences.

OK so

that we're going to switch gears and
talk about neurons for a few minutes but

that really is the presentation

as far as talking about blood cells and
suspended cells and blood and

hopefully during the presentation also you
get an appreciation for the functionality

that needs to be built into the system
this not only a simple one function type

system there's many functions that in
the end get your analysis is very much

akin to an integrated circuit where
integrated circuits have different.

Logs that allow you to
do a circuit function.

There's no question

we haven't we haven't done so studies.

You know we done these Planetary Studies
and what we do is drop out of the maybe

operate as we do plenty of studies and
then we try to find money to support

further studies and those type of
stories are the ones we want to do.

Variation between patients and
those type of studies and look

at other types of cancer for instance we
also have some more going on in head and

neck cancer and using similar approaches
to analyzing head neck cancer samples

and delivery mechanism is really the same
it's not a blood sample that we're using

to deliver it's typically suspended
cells in a buffer you know if they

were had neck cancer it would be
suspended cells in a buffer solution

really representing the state of cancer
stage so later stages of cancer.

OK So

this is talking about
neuronal Microsystems for

interfacing the neurons in this particular
system is used for kind of a special way.

There's a platform a flat platform
on which electrodes are built and

allows us to stimulate and record from the
neurons which are growing on this platform

the contribution that added to
that base platform technology.

Create this functionality on top
of this ability to record and

stimulate from these neural cultures and
how do you think about this functionality

throughout the two dimensional surface

of fluids were allowed to
be placed on the surface.

So for example this particular
configuration which is shown down at

the bottom are able to grow wherever they
want but there may be fluids that were

confined to a particular
reservoir on this platform and

a different fluid that was confined to
a different region of that platform.

So you can isolate
chemicals from one another.

Whereas the neurons were able to
grow throughout the environment.

OK All right so I'm going to flip
through a couple of these and

ask you have traumatic graph of that
system is a two compartments system and

there was growing among.

THOMPSON On the other side and looking at
the effects of the human toxin most of

this work was actually done with
investigators at Emory University.

They were interested in Parkinson's
disease and looking at the origins of

Parkinson's disease they would
use these to you know arms and

then to look and see what
the effects of applying a raw toxins

were on the distal and
they proximal end of the neurons.

So when the system going to
flip through a few of these and

someone is long time to

create passive neurons by patterning
college you know on the surface and

so we take our substrate we apply ecology

patterns to that we put the neurons
on the neurons start growing and

they'll preferentially grow
along these college and pass.

The direction lies in the growth of
the neurons from one compartment one fluid

compartment and I joining this
particular growth is with the next plan.

So I'd say it comes from the spinal cord
Durand's then growing from the next plant.

They are growing from compartment
into the next you really

can't see very well in
this particular slide but

what's happening is they're going
from one compartment to the next and

then we put this whole system inside of
a package that allows us to stimulate and

record from this culture
that's located in the center.

So again this was used for
training looking at

chemicals on either the distiller
proximal and neurons.

We also have interfacing systems that
are three dimensional so this represents

kind of an active three dimensional
scaffold for culture we know and so on.

There's towers that I have and they

also have electrodes that are located on
those kind of a three dimensional fashion

shows what they look like
in reality where this is

the fluid on the backside it's projected
that through these hollow structures.

You can see also that are on
different levels of the structure.

So it gives you this three
dimensional connectivity for

these particular systems
materials that were compatible.

When we started the research so

we had to find ways when you're in
them by compatible so we went through

many many different protocols working with
people that are very good in this area.

To come up with a protocol some example
of a protocol that has been quite hard to

render a engineers material compatible for

this particular application this
neural neuronal application

simply shows some of those
neurons growing on the tower

structures and
shows that she used for delivering.

Chemicals out of the tower.

This is the neurons which are rapidly
growing on the surface of it but

I can assure you that wasn't a case
when we started when we first took this

material and put it into a culture where
then with an hour or so we had many

many days of testing in order to go
into these materials back compatible.

So this is about compatibility studies.

We spent several years actually looking
back about abilities protect materials and

ways of statistically anyway that So

this is a lot of data say and many of
you may be familiar with those type of

we had to have a way of interfacing
these materials with cultures.

So we had a good range of scaffold for
contouring on

this package which we
put the scaffold in and

then we had fusing fluids
through the environment.

This is another view of
that same type system and

then it was put into a larger
interface and then allowed us to

interface the computers and the rest of
the signal processing side of the testing

so that was a very quick neural system so
they

include these systems for isolating fluids
as well as these three dimensional color.

During systems.

I think I'll stop with that.

Thank you.

My group leaders

working on systems that are actively
working on systems to increase throughput

so the latest systems that we're
working with they can process

around five micro leaders of blood and

about ten minutes not less
than ten minutes you will get

you can put them in parallel.

You can actually change the geometry
if you have a stronger magnet of so

if you create more magnetic force then
you can also increase the last day.

So there's a number of mechanisms that
we can use to engineering engineer

the system to have higher throughput and
that is an active area

with a new all studies we were focused

on the students were doing that work
was focused on Parkinson's disease and

he was you know we

always work on other applications of
you know with these based systems one

takes a long time to build the hard
to develop the base systems and

I assume if you see ideas for
ways they can be applied to different

cases of medicine then we're
certainly willing to and then.

Interested in talking with
you about that the questions

you know this the unit with the girl.

You know let's try to convey that you
know in addition to using that magnetic

separated from separating red
blood cells from my blood cells.

It's a new cell that's been rendered
magnetic and that includes those that have

magnetic nano particles attached to them
through receptor binding some kind of

receptor binding technique so
that I think for

this technology to be used for many
different types of studies and we were for

instance doing work with groups when
she had cancer institute where we were

looking at separation of so
those that have

nothing particular
proteins on the surface.

You know some of the C.T.

ninety seven I think I can remember all
the numbers these proteins are used in

cancer analysis but I think the CD and
I seventy seven twenty nine.

There are proteins that are expressed

when particular types of
cancer are present and so

if you can bind a nanoparticle to
the surface of those cells then you

at those receptor points then you render
those magnetic based on particular

binding of that and then you can certainly
separate those out using the system so

that I think I'm very good application for
this technology.

Maybe I'm not a blood application
where you have a team or sample.

So you have them as your systems are all
the systems is above ninety nine percent.

So the efficiency of separation is
because there's really good and

the collection of the so once they've been
separated is almost one hundred percent.

And once once we separate cells and
send them down a particular channel.

They don't stick to the side walls and

we really don't lose very
many of those cells at all.

And since you can count
the cells not in terms

of percentage probably in just
terms of raw number of cells and

I would be under ten I would say
in almost all cases for sure.

So it's it's it's a very
efficient separation mechanism

a collection mechanism so that if you
think about something like flow cytometry

you know if you take a sample and
you take a city through a flow cytometry.

First of all you know a lot of cells and
so it's not always useful for

a very small tumor samples and
if you want to separate you can count

those using a flow cytometry if you
want to use a flow cytometry to separate

the cells then you need a more cells
in order to make the process work.

So the advantage of this with respect
to flow cytometry is that and

allows you to process small volume
samples in a very efficient manner and

in ways that you can't do
it with flow cytometry

you

know we

don't you know the cancer samples or
the tumor samples that we work

with kind of cells that are up
to fifty microns in diameter.

Much larger than a normal cell.

So we have no problem working
with cells of that size with

respect to magnetic particles magnetic
particles which range range in size from

ten nanometers through maybe
two hundred nanometers diameter

just really no reason why we couldn't
use something larger for we wanted to.

So there's really no size
limitation it really depends on

the geometry of the separation
system that we build or

that we fabricate we can customize
the size for the application.