THURSDAY MORNING / CLEO 2001 / 403
2. Kersey, A.D., Davis, M.A., Patrick, H.J.,
PC
/ Electrical connection ][ = FC Connector
Optical fibre
x = Fusion splice
CThF4 Fig. 1. Schematic of system. Components are described in the text.
FFG m . mta
\ femtosecond laser pulse can be high enough to
5"
I l 438
U O
0 100 Zoo3 00 400 500 600 0 ZOO 4W 600 800
tune/sz€lxxb tine/-
FB: T e n p . mta FB; T¶u. mta
CThF4 Fig. 2. The data recorded by the seven FBG sensors for a sequence of four heating cycles of
rabbit liver-tumour hyperthermia, with increasing magnetic field strengths. In each case, the hyperther-
-
mia coil was energised from t 0 s to t 350 s. - strated that we can vaporize 10-pm diameter re-
mately known. The results indicate that the FBG
sensor has good thermal conduction to the sur-
rounding environment, good thermal isolation
between gratings, and low thermal mass (rapid
response). The effect of strain in the fibre, which
also shifts the B r a g peaks and manifests as false
temperature shifts, was not significant due to the
loose adhesion between the fibre and the liver tis-
sue.
In Fig. 3 the top four gratings are in the tu-
mour and record direct heating by magnetic hys-
teresis. The bottom three gratings, are heated in-
0 10) am m (00 .10)
directly by proximity to the hot tumour. Note
that the tumour cools rapidly due to the cooling
effect of the increased blood flow in the tumour
CThF4 Fig. 3. Density plot showing the spa-
relative to the surrounding tissue.
tial and temporal evolution of the temperature
The electromagnetic immunity and unam-
distribution during a rabbit liver-tumour hyper-
biguous spatial locations of the sensor gratings
thermia treatment. The sensors are located at the
are important for providing diagnostic measure-
labelled positions on the vertical axis. The tem-
ments for future development of human hyper-
perature distribution is interpolated between
thermia therapies, where monitoring across tu-
sensor data.
mour margins is required to ensure appropriate
differential heating between tumour and healthy
also used to provide independent temperature tissue.
measurements,and showed close agreement with 1. Webb, D.J., Hathaway, M.W., Jackson, D.A.,
the FBG temperatures, although the exact posi- Jones, S., Zhang, L, and Bennion, I., J. Bio-
tions of these point sensors can only be approxi- med. Optics, 5,45-50,2000.
I
LeBlanc, M., Koo, K.P., Askins, C.G., Put-
nam, M.A., and Friebele, E.J., 1. Lightwave
Tech., 15, 1442-1462,1997.
3. Rao, Y., Webb, D.J., Jackson,D.A., Zhang, L.,
and Bennion, I., 1.Lightwave Tech., 15, 779-
785,1997.
CThF5 11:15 am
Photodisruption in biological tissues and
single cells using femtosecond laser pulses
Nan Shen, Chris B. Schaffer, Debajyoti Datta,
Eric Mazur, Harvard University, Department of
Physics and Division of Engineering and Applied
Science, 9 Oxford St. Cambridge, M A 02138;
email: nshen0physics. harvard.edu
Semi-transparent materials such as biological tis-
sues do not strongly absorb light in the visible
range. However, the intensity of a tightly-focused
cause nonlinear absorption of laser energy by the
tissue leading to permanent material change.'
The nonlinearity of the process ensures the ab-
sorption therefore material alterations are con-
fined to the extremely small focal volume.
In this paper we show that sub-micrometer
sized photodisrupted regions can be produced
inside single cells by nonlinear absorption of
tightly focused femtosecond laser pulses. This
absorption creats a hot plasma at the focus, va-
porizing tissue. The plasma subsequently ex-
pands supersonically launching a pressure wave.'
Later, a cavitation bubble forms and eventually
c~llapses.''~The extent of the tissue damage
caused by the pressure wave and cavitation bub-
ble expansion are energy-dependent. Femtosec-
ond pulses deposit very little energy while still
causing breakdown, therefore producing surgical
photodisruption while minimizing collateral
damage.>' This suggests exciting prospects for
precise laser surgery. We have previously demon-
gions up to 100 pm beneath the surface of differ-
ent types of skin ~amples.''~Figure 1 shows two
subsurfacecavities formed by single 100-fs pulse,
20pJ of energy.
By focusing nanojoule, femtosecond laser
pulses with high numerical-aperture microscope
objectives,we cause photodisruption in cells with
sub-cellular precision. A fluorescence micro-
scope is set up colinearly with the femtosecond
.
. .
CThF5 Fig. 1. Optical microscope image of
the cross-section of a piece of mouse skin. The
two subsurface cavities in the dermis were pro-
duced using a 100-fs, 20-pJ laser pulse.
404 / CLEO 2001 / THURSDAY MORNING
laser system to visualize the target cell during
photodisruption. In our study, we use fibroblast
cells with actin fibers tagged with fluorescein
isothiocyanate(FITC).We translate the cell while
continuously irradiating to make ‘‘cutsn or we
disrupt selected regions of the cell with a varying
number of laser pulses. After laser irradiation, we
image the cell using fluorescence confocal mi-
croscopy to obtain a series of slices at different z
position, where z is the laser propagation direc-
tion. Figure 2 shows the central slice of a pho-
todisrupted cell. The cuts through the top half of
the cell are made with energy ranging from 2 to
0.5 nJ. The size of the cuts are comparable to the
width of an actin fiber bundle. In the lower half
of the cell, regions were disrupted using varying
number of pulses. Figure 3 shows the side view of
a cell cut with 2nJ, 100-fs laser pulses. Tissue is
removed only from the middle of the cell while
both the top and bottom layers remain intact.
Our technique gives us the ability to pho-
todisrupt tissues with great precision, which is
potentially useful for laser surgery. The very
small amount of energy needed and near-in-
frared wavelength of the laser light used to pho-
todisrupt sub-cellular structures suggest the pos-
CThF5 Fig. 2. Fluorescence confocal micro-
scope image of the central slice of a cell photodis-
rupted by 100-fs laser pulses. Top half of the cell
shows laser ‘‘cuts; Energy of the laser pulses used
decreases from 2 nJ at the top to 0.5 nJ at the bot-
tom. A varying number of laser pulses ranging
from 500/spot to 100/spot were used to create
“holes”in the lower half of the cell.
CThF5 Fig. 3. Fluorescence confocal micro-
scope image of the side view of a photodisrupted
cell. The cigar-shapeddark regions are laser cuts.
sibility of applying this prodedure to living cells
for cellular structure and function research.
References
1. B.C. Stuart, M.D. Feit, S. Herman, A.M.
Rubenchik, B.W. Shore, and M.D. Perry,
Phys. Rev. B53,1749 (1996).
2. E.N. Glezer, C.B. Schaffer, N. Nishimura, and
E. Mazur, Opt. Lett. Vol. 22, pp. 1917-1819,
1997.
3. P.A. Barnes and K.E. Rieckoff, Appl. Phys.
Lett. vol. 13, pp. 282-284, 1968.
4. B. Zysset, J.G. Fujimoto, and T.F. Deutsch,
Appl. Phys. B vol. 48, pp. 139-147, 1989.
5. T. Juhasz,G.A. Kastis, C. Suarez, Z. Bor, and
W.E. Bron, Lasers Surg. Med. vol. 17, pp. 1-9,
(1995).
6. B. Zysset, J.G. Fujimoto, C.A. Puliafito, R.
Birngruber and T.F. Deutsch, Lasers Surg.
Med. Vol. 9, pp. 193-204. 1989.
7. J.M. Gauss and C.A. Puliafito, “Lasers in
Ophthalmology,” Laser Surgery and Medi-
cind: Principals and Practice, C.A. Puliafito
ed. (Wiley-Liss, New York, 1996).
8. N. Nishimura, C.B. Schaffer, E.H. Li, and E.
Mazur, “Tissueablation with 100-fs and 200-
ps laser pulses”,in Proceedings of the 20th An-
nual International Conference of the IEEE En-
gineering in Medicine and Biology Society,
Vol. 20, No. 4 1703-1706 (1998).
9. N. Shen, N. Nishimura, C.B. Schaffer, A.
Brodeur, D. Datta, and E. Mazur, “Photodis-
ruption of turbid issures with ultrashort
laser pulses”, in Commercial and Biomedical
Applications of Ultrafast Lasers II, Proc. SPIE
3934,3934-24 (2000).
CThF6 a 3 0 am
Nanoshell-polymer composites for
photothermally modulated drug delivery
S.R. Sershen,J.L. West, S.L. Westcott,‘
N.J. Halas,* Rice University, Department of
Bioengineering-MS 142,6100 Main St., Houston,
TX 77005-1 892; Email: sersh@rice.edu;
*Rice University, Department of Electrical and
Computer Engineering-MS 366,6100 Main St.,
Houston, TX 77005-1892
Introduction
Optically active gold nanoshells have been incor-
porated into thermally responsive copolymers of
N-Isopropylacrylamide (NIPAAm) and acryl-
amide (AAm)for the purpose of photothermally
modulated drug delivery. The copolymer exhibits
a lower critical solution temperature (LCST) that
is slightly above body temperature. When the
temperature of the hydrogel exceeds its LCST, a
rapid collapse occurs, expelling any material con-
tained within the hydrogel. The gold nanoshells
initiate a temperature increase, via targeted ab-
sorption of near IR light.
Materials and Methods
Poly(N1PAAm-co-AAm) hydrogel disks with a
molar ratio of 95/5 ( N I P M A A m ) were fabri-
cated using the method of Dong and Hoffman.’
Gold nanoshells with a 42 nm core diameter and
a 4.5 nm thick shell were synthesized as previous-
ly described.2 The copolymer-nanoshellcompos-
ites were produced by mixing a nanoshell sus-
pension with the NIPAAm/AAm monomer
solution.
Sets of dry nanoshell-composite and control
(lacking nanoshells) hydrogels (n = 3) were
placed in a bovine serum albumin solution (BSA,
10 mg/ml) for 48 hours at 24°C. The swollen gels
were removed from the loading solution, equili-
brated for 15 minutes in 2.4 ml of 37°C Tris
buffer, and placed into a glass vial containing 2.4
ml Tris buffer at 37°C. A section of skin obtained
from the abdomen of a Hooded Long-Evans rat
was pinned over the vial after soaking in glycerol
for one hour. The hydrogels were irradiated with
a diode laser (821 nm, 2.53 W/cm2) for 40 min-
utes. Samples of the buffer were removed at set
intervals, and the amount of BSA released was
determined. An additional set of nanoshell-com-
posite hydrogels was treated as previously de-
scribed up to the start of irradiation. The hydro-
gels were irradiated for 2 minutes, followed by a
40 minute period of no irradiation. This cycle
was repeated three times, and the rate of BSA re-
leased was determined.
Results and Discussion
Nanoshell-composite hydrogels release signifi-
cantly more BSA in response to near-infrared ir-
radiation than hydrogels lacking nanoshells
(laser control) or non-irradiated hydrogels (dif-
fusion control, Figure 1). Figure 2 shows that
nanoshell-composite hydrogels can deliver mul-
tiple bursts of BSA in response to cyclic irradia-
tion.
The interaction between incident near in-
frared light and the nanoshells heats the copoly-
mer, forcing the hydrogel to collapse. The col-
lapse initiates an increase in the release rate of a
model drug, as shown in Figure 1. Figures 1 and 2
demonstrate photothermally modulated drug
delivery in a transdermal in vitro model. Taken
together, these results indicate the possibility of
f ‘20
--
- =$= &
:
E
Diffusion Control
I
$
1
0 IO 20 30 U)
Time (mh)
CThF6 Fig. 1. (Left) BSA release from
nanoshell-composite and control hydrogels in
response to near-infrared irradiation (821 nm,
2.53 Wlcm‘).
I I I I I I I
V I
-20 0 20 a M) 80 100 120
Time (mh)
CThF6 Fig. 2. (Right) Multiple bursts of BSA
release from nanoshell-composite hydrogels in
response to cyclic near-infrared irradiation (821
nm,2.53 W/cm2).