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Surface Science 500 (2002) 656–677

www.elsevier.com/locate/susc

Biological surface science


Bengt Kasemo *
Department of Applied Physics, Chalmers University of Technology and G€oteborg University, S-412 96 G€oteborg, Sweden
Received 13 December 2000; accepted for publication 5 July 2001

Abstract
Biological surface science (BioSS), as defined here is the broad interdisciplinary area where properties and processes
at interfaces between synthetic materials and biological environments are investigated and biofunctional surfaces are
fabricated. Six examples are used to introduce and discuss the subject: Medical implants in the human body, biosensors
and biochips for diagnostics, tissue engineering, bioelectronics, artificial photosynthesis, and biomimetic materials.
They are areas of varying maturity, together constituting a strong driving force for the current rapid development of
BioSS. The second driving force is the purely scientific challenges and opportunities to explore the mutual interaction
between biological components and surfaces.
Model systems range from the unique water structures at solid surfaces and water shells around proteins and
biomembranes, via amino and nucleic acids, proteins, DNA, phospholipid membranes, to cells and living tissue at
surfaces. At one end of the spectrum the scientific challenge is to map out the structures, bonding, dynamics and ki-
netics of biomolecules at surfaces in a similar way as has been done for simple molecules during the past three decades
in surface science. At the other end of the complexity spectrum one addresses how biofunctional surfaces participate in
and can be designed to constructively participate in the total communication system of cells and tissue.
Biofunctional surfaces call for advanced design and preparation in order to match the sophisticated (bio) recognition
ability of biological systems. Specifically this requires combined topographic, chemical and visco-elastic patterns on
surfaces to match proteins at the nm scale and cells at the micrometer scale. Essentially all methods of surface science
are useful. High-resolution (e.g. scanning probe) microscopies, spatially resolved and high sensitivity, non-invasive
optical spectroscopies, self-organizing monolayers, and nano- and microfabrication are important for BioSS. However,
there is also a need to adopt or develop new methods for studies of biointerfaces in the native, liquid state.
For the future it is likely that BioSS will have an even broader definition than above and include native interfaces,
and that combinations of molecular (cell) biology and BioSS will contribute to the understanding of the ‘‘living
state’’.  2001 Elsevier Science B.V. All rights reserved.

Keywords: Adhesion; Biological compounds; Biological molecules – nucleic acids; Biological molecules – proteins; Solid–liquid
interfaces

1. Introduction

Imagine the following six situations where the


*
Tel.: +46-317723370; fax: +46-317723134. properties of solid surfaces are or may become
E-mail address: kasemo@fy.chalmers.se (B. Kasemo). important in practice:

0039-6028/01/$ - see front matter  2001 Elsevier Science B.V. All rights reserved.
PII: S 0 0 3 9 - 6 0 2 8 ( 0 1 ) 0 1 8 0 9 - X
B. Kasemo / Surface Science 500 (2002) 656–677 657

(i) A patient with severely degraded dental sta- (iii) A small number of living cells of a partic-
tus is treated by a surgical procedure, where one or ular kind, maybe a tissue culture or so-called stem
several dental implants, made from metal or ce- cells for a certain type of human tissue, are placed
ramic, are implanted into the jawbone so that they in a scaffold made from some synthetic mate-
after some healing in period can function as arti- rial, with the intention to make the cells grow in
ficial teeth (Fig. 1). Other types of implants are number and differentiate ex vivo into a new func-
shown in Fig. 2. tional tissue, which later is placed in a patient, in
(ii) A blood or urine sample is distributed over a order to repair a lost or degraded body function
suitably prepared biosensor or biochip surface in (Fig. 4).
order to diagnose a patient’s health status or al- (iv) Neural cells (neurons) are placed on a
ternatively for a forensic identification (Fig. 3). micropatterned surface, where they self-organize
into a functioning neural network, which can be
addressed chemically and/or electronically by in–
out (I/O) connections so that a cell-based bioelec-
tronic circuit is achieved.
(v) A particular kind of photosensitive, charge
transfer proteins are attached to and organized on
a specially designed material surface in order to
harvest the energy of sun light with a high effi-
ciency, thereby converting the light into chemical
or electrical energy in a process mimicking the
photosynthetic process of green plants or certain
bacteria.
(vi) A surface is microfabricated with an array
of specially architectured, soft protrusions in the
micrometer size range, mimicking a shark or dol-
phin skin and thereby providing a dramatic re-
duction in hydro- or aerodynamic friction.
The medical implant example [1,2] is a clinical
reality since many years and hundreds of thou-
sands of patients have been treated with a major
increase in life quality. There are many other ex-
amples of medical implants (Fig. 3), each one un-
iquely different from the others in the biological/
clinical details e.g. artificial hip and knee joints,
artificial blood vessels and heart valves, and syn-
thetic intraocular lenses. Together they represent a
large number of industries with total turnover
approaching or even exceeding a hundred billion
dollars per year.
The biosensor example [3–5] is representing an
area, where commercial products already exist
both for single sensors and array type sensors [6]
(e.g. so-called DNA chips) which are used clini-
Fig. 1. Schematic illustration of the interface between a dental
cally or at the R&D level in biomedical industries
implant and the jawbone into which it is implanted, at different
magnifications. After the surgical procedure the surface is first and academia. Figs. 4 and 5 illustrate some of the
exposed to water, then to proteins, and eventually to cells (see current sensing principles and surface immobili-
Fig. 7). See Fig. 2 for other types of implants. zation strategies. This area is in extremely rapid
658 B. Kasemo / Surface Science 500 (2002) 656–677

Fig. 2. A selected set of different medical implants.

Fig. 3. Some common detection principles for biosensors and biochips.


B. Kasemo / Surface Science 500 (2002) 656–677 659

Fig. 3 (continued)

Fig. 4. Tissue engineering scaffold. A template of a synthetic material forms the ‘‘host’’ for a cell or tissue culture. The scaffold should
promote the development of the culture into a functioning tissue.
660 B. Kasemo / Surface Science 500 (2002) 656–677

Fig. 5. Immobilization strategies for biomolecules on surfaces to achieve biosensor and biomedical diagnostic functions.

development with an estimated market growth for e.g. for real-time control of drug administra-
labchips/bioarrays of 40% per year. Most of its tion. One such example is the search for a reliable
impact, technically and commercially, is yet to glucose sensor for real-time control of insulin
come in the wake of the so-called molecular bio- administration for diabetes patients, via an im-
logy or biotechnology revolution. The product will planted, sensor controlled insulin pump.
have important input from modern microelec- Tissue engineering [7,8] is an emerging area
tronics, materials and surface science, advanced where most of the impact is also still to come, very
micro- and nanofabrication, and bioinformatics. much fueled by the molecular and cell biology
The perspective for the future is unprecedented revolution. The basic strategy for repair of lost
precision and speed in multiple (broad band), and body functions by way of tissue engineering is dif-
individualized clinical and eventually ‘‘near pa- ferent compared to medical implants: In the latter
tient’’ (home) diagnostics. Other important appli- case a synthetic component is fabricated that re-
cations are expected in e.g. environmental control places the function of the lost or degraded part of
and food production. the body, such as the metal ball head on the hip
Although the focus up till now has been pri- implant, and a matching counterpart made from
marily on DNA-based diagnostics by large sensor high molecular weight polyethylene (HMWPE),
arrays, the future sensing arrays will also address constituting an artificial hip joint. In the tissue
biologically more ‘‘down stream’’ components in engineering strategy the idea (Fig. 4) is instead to
the cell expression and cell–cell communication produce a real functioning tissue from a cell cul-
processes, typically peptides and proteins (the lat- ture ‘‘seed’’, by making it grow ex vivo in a suit-
ter area is sometimes referred to as ‘‘proteomics’’). able environment. The latter requires a synthetic
Even further down the line lies (potentially) cell- scaffold or template for the growing cells/tissue,
based diagnostics (cellomics). whose macroscopic geometry, microscopic topo-
Another type of sensors is the kind that can be graphy and surface chemical properties cooperate
implanted and used in feed back systems in vivo with other stimuli such as extracellular signal
B. Kasemo / Surface Science 500 (2002) 656–677 661

substances and the composition of the physiolog- have new unique characteristics originating from
ical solution etc., to make the cell culture develop the biomolecules’ highly specialized functions, in-
into a functioning tissue, ready for implantation. cluding their replication and recognition ability,
Practical tissue engineering is today realized only and self-organization. The cell level, reverse engi-
for a few ‘‘simple’’ tissues like skin and, to some neering approach will result in entirely new circuit,
extent, cartilage, while large research efforts are component, and functional concepts. The idea is
focussed on e.g. liver, pancreas, blood vessel, to ‘‘automatically’’ achieve signal- and informa-
bladder, and heart valve tissues. tion-storage pathways from the internal self-
Bioelectronics [9–11] is still only at the very organization, architecture and multiple functions
early, explorative research stage and it is too early of living cells, and from the biosystem’s inter- and
to judge if it will or will not become a practical intracellular communication pathways. The latter
reality in the future. The basic idea is to take ad- have evolved over billions of years, and thus one
vantage of some unique aspects of nature’s own does not need to invent these functions and ar-
way of information storage and processing. This chitectures again (the latter is the prime reason for
can alternatively be done at the cellular level calling this approach reverse engineering).
(sometimes referred to as ‘‘reverse engineering’’; As in the bioelectronics case, biomimetic or
see below), where the primary building stones are artificial photosynthesis [12] is still an area at its
networks of real cells, or by ‘‘forward engineering’’ infancy. The basic mechanism is known by prin-
at the biomolecular level, where the building ciple (Fig. 6); it is based on electron excitation by
stones are peptides, proteins, DNA strings etc., incident, photons in suitable proteins or protein
and combinations of them. Both in the cellular and arrays, followed by charge separation (preventing
molecular approach the biological components are charge recombination and concomitant energy
likely to be steered into a suitable pattern in or on loss), and finally redox reactions creating useful,
a microengineered template or substrate surface, energy rich (fuel) molecules. However, biomimetic
providing chemical and/or electrical I/O commu- structures that have the quantum efficiency, sus-
nication. Bioelectronic components and circuits tainability, and a self-repair ability approaching
made by the molecular level, forward engineering those of natural systems, and that can produce
approach are likely to resemble today’s micro- useful fuels (like hydrogen) are still just a vision
electronics circuits in some respects, but will also and goal for future research. A strong driving

Artificial Photosynthesis

LUMO
e- Re

e-
Catred A-
e- Electron
energy
A (Qualitative)
(Acceptor)

D+
PIGMENT
D e- Catox

(Donor) e- Rh
e-
HOMO

Re = Electron Relay
Rh = Hole Relay

Fig. 6. The principle of photon to electron energy conversion in photosynthesis.


662 B. Kasemo / Surface Science 500 (2002) 656–677

force is the rapidly growing need for sustainable living cells) are briefly discussed. The same para-
energy systems. One strategy to realize such struc- graph also discusses different types of surfaces and
tures is to immobilize the photon-harvesting and surface modifications, and analytical and prepa-
redox protein arrays of real plants, or their syn- ration methods for biofunctional surfaces. How-
thetic counterparts, on surfaces with suitable ever, these paragraphs are intentionally very short,
chemical and electronic properties. These surfaces since the focus is on general aspects and concepts
need to be functionilized both with respect to rather than specific surfaces or methods. Finally
nano- and microtopography and with respect to some comments are attempted regarding the likely
surface chemistry. Another approach is to build future development of BioSS.
entirely synthetic nanostructures mimicking the The intention with the article is not to present a
photosynthetic systems. comprehensive review of the field, but rather to
The low friction mimic of a sharkskin surface identify some important trends, directions, and
constitutes just one example of biomimetic mate- concepts in BioSS and particularly to outline the
rials science [13]. In biomimetic materials science opportunities for graduate students and surface
the idea is to mimic the functional properties of scientists to contribute to this exciting field. Ref-
biological materials/components or the processes erences are therefore chosen primarily to provide
by which they are manufactured in nature, in order starting points for further reading, and there is no
to achieve new, superior materials properties or ambition to cover all previous work in BioSS (i.e.
manufacturing advantages. A major driving force the reference lists in the quoted references are just
to attempt mimicking the sharkskin is the poten- as important as the textual content).
tially much lower energy consumption of airplanes
and sea vehicles that would be obtained. A more
exotic example, already realized in real products, is 2. The role of surfaces
swimsuits for faster swimming and low friction
hull coatings on sailboats. Other biomimetic ma- 2.1. Biorecognition
terials examples are antifreeze materials, utilizing
the special features of some water holding proteins In most of the examples above, especially for
that can lower the freezing point of water in cer- (i)–(iv), biorecognition is a central component. Any
tain arctic fishes below 0 C, and self-cleaning attempt to make a sophisticated, functional sur-
surfaces like some green leaves (the lotus leaf), and face for biointeractions must take into account the
self-repairing surfaces. highly developed ability of biological systems
to recognize specially designed features on the
1.1. Biological surface science molecular scale. Famous examples are antibody–
antigen, enzyme–substrate, and receptor–trans-
In all six examples above, each representing mitter recognition (e.g. in cell membranes). The
large and growing fields of applications and asso- recognition is programmed into the molecules
ciated basic R&D, surfaces play a very important through the combination of their 3D topographic
role. As a generic name for this branch of surface architecture, the superimposed chemical architec-
science I use biological surface science (BioSS) [14] ture, and the dynamic properties. Consequently an
(it includes the sub-area of medical applications; optimally designed surface for specific biological
biomedical surface science). function must take these aspects into account.
The importance and function of synthetic solid Although the fundamental interactions occur
surfaces in biological and medical contexts, is the on the molecular scale there is an interesting and
subject of the next paragraph (Section 2), with unique synergistic connection between the na-
special focus on the six examples or sub-areas nometer and the micrometer length scales [1,15,16]
mentioned above. In Section 3 the large variety of when cells are present, as e.g. in the cases of
model systems available for BioSS studies, of dif- medical implants, tissue engineering and cell-based
ferent complexity (from water and amino acids to bioelectronics. This is schematically illustrated in
B. Kasemo / Surface Science 500 (2002) 656–677 663

Fig. 7. Schematic illustration of the successive events following after implantation of a medical implant. The first molecules to reach
the surface are water molecules (ns time scale). The water shell that is formed affect the protein interaction starting on the micro- to
millisecond time scale, and continuing for much longer times. The water shell on the surface affects the protein interaction. Eventually
cells reach the surface. Their surface interaction takes place via the protein coating whose properties is determined by the surface and
water adlayer properties.

Figs. 7 and 8. The first picture (Fig. 7) shows the may determine if proteins denature or not, their
sequence of events after a surface has suddenly orientation, and coverage etc.
been placed in a biological milieu containing cells. When cells arrive at the surface they ‘‘see’’ a
The first molecules to reach the surface (time scale protein-covered surface whose protein layer has
of order ns) are water molecules. Water is known properties that were initially determined by the
to interact and bind very differently at surfaces preformed water shells. Thus, when we talk about
depending on the surface properties (see Section cell–surface interactions, it is ultimately an inter-
3). The properties of the surface water ‘‘shell’’ are action between cells and surface bound proteins
an important factor influencing proteins and other (or other biomolecules). The latter is illustrated in
molecules that arrive a little later. These water- Fig. 8, where the clean surface in the illustration is
soluble biomolecules also have hydration (water) deliberately covered with a native or artificial
shells and the interaction between the surface biomembrane (supported bilayer), containing em-
water shell with the biomolecular water shell in- bedded receptors that can specifically interact
fluences the fundamental kinetic processes and the (bind to and/or provide I/O signals etc.) with cells
thermodynamics at the interface. For example, it approaching the surface.
664 B. Kasemo / Surface Science 500 (2002) 656–677

only marginally applying surface analytical tech-


niques to study medical implant surfaces, to a
current situation where such use is the rule and
mandatory for high quality research, and also for
legal control, standardization and quality control
of commercial products. Let us briefly inspect how
surfaces come into play.
For a dental implant (Fig. 1), which is just one
of many examples of so-called bone anchored
implants, the clinical goal is to obtain a long term
(i.e. life long) secure anchoring of the implant. The
latter includes e.g. the implant’s ability to carry
and sustain the dynamic and static loads that it is
subject to. It is obviously (for the patient and for
cost reasons) important to achieve this function
with the shortest possible healing time, with a very
small failure rate and with minimal uncomfort
for the patient. The implant is put in place after a
surgical procedure typically involving drilling a
hole in the cheekbone, threading of the hole, and
then placing the screw-shaped end of the implant
into the threaded hole.
The surface design is one of many components
contributing to the clinical goal fulfillment. An
inappropriate material choice or surface coating
might lead to a too strong inflammatory response
that adds on top of the already initiated inflam-
matory process by the surgical procedure. Then
Fig. 8. A conceptual approach to convert a synthetic surface to connective tissue rather than bone is formed
a biomimetic surface by coating it with a supported biomem- around the implant, with concomitant loss of
brane with built-in functional membrane-bound proteins. mechanical stability and eventual implant failure.
A less perturbing surface may lead to a bone
healing process, which is similar to the one in the
Figs. 7 and 8 emphasize the above mentioned absence of an implant i.e. as if the implant were a
large size range that the functional units in biology fairly passive spectator (sometimes this is referred
cover, from water molecules and small proteins at to as inert materials or bone accepting materials,
sub-nanometer sizes, via cell membranes and su- although no material is really inert in an absolute
pra-molecular complexes in the 10 nm range, to sense). Some surface coatings appear to have the
cells in the micrometer range and finally fully or- ability to induce bone formation, for example the
ganized tissue and organs at the macroscopic size calcium phosphate materials usually referred to as
level. hydroxyapatites.
More recently there has also been attempts to
2.2. Medical implants speed up the bone healing beyond the normal
healing rate by adding so-called bone growth fac-
In the case of medical implants (Figs. 1 and 3) tors. The latter can e.g. be implemented by using a
the importance of surface science is quite obvious porous surface with a slow release of the desired
[1,2,14–16]. In 15–20 years time biomaterials re- biomolecules. Engineering of such surfaces re-
search and development has transformed from quires a whole set of surface science tools. Exam-
B. Kasemo / Surface Science 500 (2002) 656–677 665

ples of used surface science methods are coating that ‘‘detector molecules’’ are attached/immobi-
technologies to deposit the desired surface material lized on a solid surface in such a way that a specific
on a bulk carrier; surface spectroscopies like XPS, signal is obtained from the sensor when the de-
AES, and SIMS to control e.g. the Ca/P ratio; tector molecules selectively react with (bind to) the
SEM or scanning probe techniques (SPMs) to biomolecules they are designed to detect. In other
control surface structure and chemistry, both of words, the detection relies on the biorecognition
which influence the biological response; micro- between pre-designed native or synthetic reagent
and nanofabrication or other techniques to make molecules and unknown sample molecules. The
porous surfaces; surface impregnation by mole- biorecognition event typically occurs when the pre-
cules that are released at a pre-programmed rate to coated sensor is exposed to blood or urine samples
promote healing/bone formation, FTIR to mea- or extracts thereof.
sure adsorbed biological molecules etc. The detector read-out signals may be electrical,
Another challenging task for surface engineer- optical, or some other signal that is convertible
ing of biomaterials is blood-compatible materials, into a measurable electrical signal. The detection
such as the inner walls of artificial blood vessels, may occur in real time, i.e. just when the recog-
and the blood contacting surfaces of heart valves, nition event occurs, or the sensor can be read-off
dialyses equipment, and heart–lung machines. at a later time. Two common examples are (i)
Normal blood is, naively speaking, a metastable antigens are pre-adsorbed on the sensor surface to
state stabilized between two opposing driving detect the corresponding antibody(ies) in a clinical
forces––coagulation and anticoagulation. Almost sample for diagnostics; (ii) a single strand DNA
any external perturbation in the form of an arti- segment (i.e. a nucleic acid chain without cross-
ficial surface tends to trigger the coagulation sys- linking), representing a specific, known part of the
tem, i.e. blood clotting may occur. The extent to genetic code, is attached to the surface so that all
which this happens, and is detrimental for the the dangling bonds of the non-hybridized, single
patient, depends on a number of factors such as strand are exposed and available for binding to a
where the surface is placed in the blood stream, the matching DNA strand. When the sensor is ex-
local hydrodynamics, the flow speed, contact time posed to a DNA sample to be analyzed, one wants
between the surface and blood and, of course, on to obtain a specific response when there is per-
the surface chemistry. fect matching between the detector molecule and
There is today clear evidence that the latter has some sample molecules, while a different response
a major influence on blood compatibility and that should be obtained when the matching is imper-
surfaces can be engineered to suppress blood co- fect. On a DNA chip many different such DNA
agulation, at least for considerable time. However, segments are placed at different locations and thus
there is still a long way before surfaces that mimic many different DNA segments can be analyzed in
the blood compatibility of the endothelial cells of one ‘‘shot’’ (bioarray, biochip). When the sensor
normal blood vessel walls are achieved. For the array is combined with a sophisticated microflui-
surface scientist this task involves designing du- dics system often utilizing functional hydropho-
rable surface coatings that have the right chemical bic–hydrophilic surfaces––we call the device a
and micromechanical (visco-elastic) properties, lab-on-a-chip or just labchip.
which in turn may require a better understanding Other sensing principles are to e.g. employ en-
than today of how the native system works, so that zymes that are immobilized on a surface with
a true biomimetic approach can be implemented. preserved function, and which can recognize spe-
cific biochemical substances, or the use of MAL-
2.3. Biosensors and biochips DI-MS to identify proteins and antibody–antigen
complexes.
A variety of surface based detection principles Surface science tools and expertise come into
are employed for biosensors/biochips (Figs. 2 and play in many ways in the preparation and use of
3). A common principle of a biosensor/biochip is biosensors and biochips. Immobilization of the
666 B. Kasemo / Surface Science 500 (2002) 656–677

detector molecules in such a way that they preserve material (Fig. 4), on which the original cell cul-
their original recognition specificity, requires sur- ture(s) are deposited and eventually develop into a
face pre-coatings that simultaneously allow suffi- tissue, must be specifically designed to promote,
ciently strong binding of the detector molecules, and not compromise, the tissue evolution. This
and sufficiently weak and gentle perturbation that calls first of all for a non-toxic surface for the cell
the molecules retain their functionality (Fig. 5). culture, and for suitable protein adsorption prop-
For example, the strong polarization forces at erties, since the protein adsorption layer that
metal surfaces and the strong ionic or covalent always forms on the surface influences the cell–
interactions on many inorganic metal oxides and surface interactions. Since differentiated tissue is
semiconductor materials may cause denaturation an ultimate goal, different parts of the scaffold may
and loss of specificity of biomolecules. At the same require different surface functionalities, i.e. differ-
time such materials may be desirable ‘‘platforms’’ ent surface chemistries and topographies.
for sensor arrays. A solution is then deposition of Cells in a tissue culture communicate with each
special spacer and/or linker molecules (typically other by sending out and receiving signaling mol-
some type of polymer or a native or synthetic ecules to/from the extracellular matrix. The total
biomembrane) on the original sensor substrate. spectrum of such signals is influenced by the sur-
The preparation and characterization of such faces onto which cells attach. Thus, the surfaces of
layers for useful deposition of detector molecules, tissue engineering scaffolds are an integral part of
obviously require sophisticated surface science in- the communicating and self-organizing tissue.
put. The same holds for the preparation of the part A common––but not the only one––concept in
of the sensor that provides a measurable signal. tissue engineering is a scaffold that eventually is
The most common detection approaches so far are dissolved as the tissue ‘‘matures’’. This calls for a
based on optical detection in the visible or infra- sophisticated ‘‘programming’’ of the material so
red. Optimization of the optical detection requires that it dissolves into harmless products at the right
deposition of optically suitable layers e.g. regard- pace, and maybe also exposes different functional
ing reflectivity, absorption properties, plasmon surfaces at different times. (The latter may also be
properties, field enhancements etc., which are all a desired function of medical implants.) Many
typical components in thin film design of optical different surface technologies are potentially suit-
coatings. Consequently advanced surface spec- able for such surface functionalization, especially
troscopy and microscopy come into play both for coating technologies that can be applied to 3D
understanding of how the matrix and detector curved surfaces and porous surfaces, but also the
molecules bind to the sensor surfaces and for standard surface characterization methods come
practical diagnostics. Regarding biochips, arrays into play. The 3D surface topography, which is
of detector ‘‘spots’’ are required and here surface very important for cell–surface interactions, may
science plays a similar role as it does for the mul- require micro- and nanofabrication methods.
tiple microfabrication steps in semiconductor/mi-
croelectronics technology. 2.5. Bioelectronics

2.4. Tissue engineering Bioelectronics based on living cells (neurons


etc.) call for patterned surfaces where the cells can
To reach the goal of tissue engineering––to be confined to certain geographic locations (in
grow a functional tissue ex vivo that can later be order to make a reproducible circuit), and where
implanted into a human body––requires control they can be kept alive and functional, which in
over a large number of external parameters. Ex- turn requires suitable surface chemistries and to-
amples are the nutrients, extracellular signal sub- pographies. Regarding the basic control of cells
stances, antibiotics, and the dynamic mechanical there is actually large similarities between cell-
force fields etc. in the bioreactor where the tissue is based electronics and tissue engineering. The sur-
grown. The surface of the template or scaffold face patterns for bioelectronics require in addition
B. Kasemo / Surface Science 500 (2002) 656–677 667

adequate I/O connections for electrical or chemi- amples involving surfaces as key elements are
cal communication. Similar requirements are at mimics of the lubrication and wear resistance of
hand for bioelectronic devices based on biomole- artificial joints, the low drag friction of shark and
cules rather than whole cells, albeit at a generally dolphin skin, the self-cleaning action of some
smaller length scale (see also the text under artifi- green leaves, and threads made to mimic the
cial photosynthesis below). Consequently bioelec- strength and flexibility of spider web.
tronic circuits require a combination of molecular
cell biology with surface preparation/character-
ization and micro- or nanofabrication techniques. 3. Model systems, research topics, and ongoing
research
2.6. Artificial photosynthesis
3.1. Biological model systems
From a surface design point of view there are
many similarities between biomolecule-based bio- In this section model systems are discussed and
electronics and artificial photosynthesis based on brief references are made to recent or ongoing re-
biomolecules. One common denominator is charge search, illustrating the diversity of research areas
transfer within or between biomolecules, and be- and the challenging research problems in BioSS.
tween biomolecules and surfaces, either induced by The biologically relevant model systems available
external electrical fields (voltage bias) or by the for surface science approaches cover such a broad
electromagnetic field of light in the visible regime range that any taste should find its topic of choice.
(e.g. photoexcitations of electron hole pairs). The Some of these areas––such as water and simple
relationship between the two areas becomes espe- amino acids––are immediately approachable by
cially obvious when the bioelectronics concepts are existing UHV- and surface science preparative and
extended towards opto(bio)electronic devices. analytical techniques. Others require entirely new
For the surface scientist a major contribution or modified experimental approaches, such as
can be to produce chemical–topographic patterns those requiring wet preparation and in-liquid an-
on surfaces that adsorb and retain the biomolec- alyses or e.g. freeze-drying techniques, to maintain
ular building blocks of the circuit elements at the biological properties and function of the
prescribed locations and in the right orientations studied systems. The model systems contained in
(self-organization), without loss of functionality. the following list should be read with the accom-
In artificial photosynthesis with e.g. proteins (Fig. panying label ‘‘... at surfaces’’:
6) there is also a major challenge to find and pre-
pare surfaces that match the electronic states of the • water
charge transfer proteins, e.g., as electron/hole ac- • amino acids, nucleic acids, lipids
ceptors or donors. Further requirements of the • peptides, DNA segments
surface may be to make field enhancement struc- • proteins
tures that increase the quantum efficiency of • DNA
‘‘photon harvesting’’; perhaps similar to the ones • lipids, biomembranes
that produce the recently discovered giant surface • cells
enhanced Raman effect. Other ideas that come • tissue, in vitro
into play are to use optically active quantum dot • tissue, in vivo
arrays to catch and convert photons to energetic • microorganisms
and ‘‘useful’’ electrons.
The list covers most but not all biointerfaces
2.7. Biomimetic materials where there is an interface between a synthetic (i.e.
non-native) material surface and some component(s)
This heading covers a large number of widely of native biological systems. I call them hybrid
different scientific and technical applications. Ex- biointerfaces, in contrast to native biointerfaces,
668 B. Kasemo / Surface Science 500 (2002) 656–677

because they consist of one native biological rent progress in the modeling of water hydration
component and one synthetic, non-native com- shells on protein resistant self-assembled mono-
ponent. I will not treat native biointerfaces, layers by Gruntze and co-workers [19], which is an
such as cell–cell or microorganism–cell interfaces, important component both for understanding of
although they are closely related to the hybrid what the requirements protein resistant surface is,
interfaces; it is actually very likely that the and for understanding protein adsorption. It is
knowledge and methods for these two types of also a central component for blood-compatible
interfaces will mutually interact and cross-fertilize surfaces.
in the future. For example, studies of properties
and processes in real cell membranes and in so- 3.1.2. Amino acids and peptides
called supported biomembranes (see below) are Amino acids are the building stones of one of
clearly benefiting from each other. the most elementary functional units in biology,
namely proteins. Mapping their interactions with
3.1.1. Water surfaces is therefore, apart from the inherent basic
Water at interfaces is an area where surface research interest, a precursor to the understanding
science studies will contribute significantly in the of how peptides and proteins interact with sur-
future, e.g. to the understanding of what is some- faces. In the recent years a number of papers have
times called ‘‘biological water’’. The latter refers to been published [20–24] addressing how sub-
the special water structures that are formed near monolayer to multilayer deposits of simple amino
surfaces or at narrow interfaces between biological acids (glycine, alanine, . . .) interact with single
components, such as the hydration shells of pro- crystal surfaces or polycrystalline surfaces in
teins, DNA and biomembranes. Biological water vacuo. These studies include standard spectro-
at surfaces, or in small confined volumes, is in scopic techniques, such as SIMS, HREELS, FTIR,
principle closely related to the special water and synchrotron light based spectroscopies applied
structures that form at solid synthetic surfaces. to amino acid adsorbate layers. Some of these
They obtain their unique interfacial properties systems have also been characterized by LEED
through a combination of the local physical– and TDS. There are also recent attempts to de-
chemical interactions (the thermodynamics) at the scribe the amino acid–substrate interactions (in-
interface, including the spatial constraints caused cluding adsorbate–adsorbate interaction) by first
by the surface, and the kinetic constraints. They principle calculations.
are also related to the hydration shells of solvated The amino acids––at least the simpler ones––are
ions and to the Helmholtz layers at electrode sur- compatible with UHV evaporation techniques.
faces in electrochemical cells, an area where sur- For example glycine and several other amino acids
face science has contributed with significant new can be evaporated from a simple Knudsen source
insight in the past two decades. at low temperature. More complex (fragile) amino
Since the presence of water has such a profound acids may require deposition from the liquid
influence on both the thermodynamics and kinetics phase, and then similar techniques are needed
at biointerfaces it is a safe prediction that it will be as have been developed for UHV studies of elec-
a major topic in BioSS for a decade or more ahead. trode double layers in electrochemistry. Important
Recent conceptual and theoretical considerations questions concern (i) how the amino acids bind to
can be found in the series of papers published by the surface, if the bonding is non-dissociative or
Vogler and collaborators (see Ref. [17] and refer- dissociative, where and how the bonds are estab-
ences therein). Experimental work includes both lished (at the amine group, and/or at the car-
direct studies of the force interactions at hybrid boxylic group, and/or at the R-side group etc.), (ii)
interfaces, as described by e.g. Israelachvili and strength and character of adsorbate–adsorbate
collaborators [18], and kinetic and spectroscopic interactions, (iii) molecular orientations, (iv) 2D
studies of interfacial water, using both biological crystal structure (when ordered layers are formed),
and non-biological substances. There is also cur- and (v) the influence of water molecules on these
B. Kasemo / Surface Science 500 (2002) 656–677 669

properties. Another class of questions regards ki- is gradually covered by a single protein type or by
netic processes, e.g. to what extent can surfaces proteins that compete for the surface sites. Com-
catalyze polymerization (e.g. forming peptide mon techniques are surface plasmon resonance,
bonds) and other reactions between amino acids ellipsometry, fluorescence detection, and other
and co-adsorbates. optical methods, and gravimetric (QCM, QCM-D,
The same questions as above can be formulated SAW-devices) and radiolabelling methods. Pro-
for peptides, which are polymer chains of amino tein adsorption is for most surfaces irreversible
acids. They are important functional units in through additive contributions from e.g. van der
biology per se, and also constitute the amino-acid- Waal’s interaction, polar and charged groups on
chain sub-units of proteins. Studies of peptides are the proteins, hydrogen bonding, and hydrophobic
therefore one step up in complexity from amino interactions.
acids, but also a step closer towards ‘‘real bio- The relative strengths of these contributions
logy’’. (By analogy the step from single nucleic depend on protein type, on surface properties and
acids at surfaces to DNA segments at surfaces is a on pH and salt concentration. Binding energies are
step both in complexity and towards biological difficult to measure for irreversible adsorption. A
relevance.) A natural extension of studies of amino few surfaces bind proteins sufficiently weakly that
acid monolayers is thus di-peptide, tri-peptide reversible adsorption occurs, whereby heats of
etc. adsorption studies. Up to date no systematic adsorption can be obtained from the kinetic mea-
peptide studies, like the ones for amino acids, have surements.
to my knowledge been done in UHV, with state of Very few surfaces are so-called protein resis-
the art surface science techniques. In contrast there tant surfaces (see also under biomembranes be-
is large ongoing activity focussing on liquid phase low). Protein resistance means that the adsorbed
deposition of various peptide depositions and amount, even at relatively high protein concen-
formulations on biomaterial surfaces in order to tration in the bulk liquid, is below or near the
make them more functional for e.g. medical im- detection limit. Theoretically it is still a major
plant and tissue engineering applications. These challenge to understand the mechanisms that con-
depositions still lack the rigorous control that can tribute to protein resistance of surfaces, and even
be achieved in UHV, but surface spectroscopies more so to understand the detailed contributions
like XPS and FTIR can and are applied, after to strong(er) protein–surface bonds. Recent ex-
deposition, to control the deposited layer compo- perimental work by Whitesides and co-workers
sition. [40–42] and by Gruntze and co-workers [19,39]
constitute important steps towards understanding
of protein adsorption, and protein resistance and
3.1.3. Proteins ultimately of protein binding more generally.
Proteins, which are (polymerized) peptide Important challenges, not at all clarified at the
chains wrapped folded into special 3D structures molecular level, are to quantify the relative and
(conformations), have since long been, and is absolute contributions of the various protein–
currently the subject of extensive surface studies surface interaction terms, and to ultimately
[25–42] due both to their central role in all bio- understand how the relatively fragile protein
logical processes and due to their importance in structures (usually they denature well below 100
the context of bioengineering, e.g. for biosensors, C) are affected by the perturbation exerted by a
biofouling and tissue engineering. The vast ma- solid surface. There is no doubt that the water shell
jority of past studies have focussed on the (mac- of the surface and that of the (water soluble)
roscopic) kinetics, and to some extent on the proteins, and their mutual interaction, play an
energetics of adsorption on various surfaces, and important role, which can be expressed in terms of
under different conditions of liquid properties hydrophobicity–hydrophilicity at the macroscopic
(protein concentration, pH, salinity, . . .). A typical scale, and in terms of water–water versus water–
experiment records the kinetics by which a surface surface bonding strengths at the molecular level.
670 B. Kasemo / Surface Science 500 (2002) 656–677

For example, a material surface of low polariz- techniques are already addressing some of these
ability in the bulk with a strongly bound water questions. There are major challenges to adopt
mono- or bilayer on the surface, is likely to only these techniques to the special ‘‘wet’’ requirements
weakly perturb a protein with predominantly hy- for proteins. The theoretical work ahead is equally
drophilic surface domains, while a hydrophobic demanding and important as the experimental
surface is likely to strongly perturb a protein with work and will be absolutely necessary for progress
mixed hydrophilic and hydrophobic domains, via in this area.
hydrophobic interactions with the latter. These
ideas have recently been quantified theoretically in 3.1.4. DNA
some simple cases. Furthermore there are impor- Studies of the DNA molecule at surfaces [44,45]
tant entropy effects in the energetics of protein is driven both by the inherent interest in under-
bonding at surfaces due to the highly organized standing different aspects of this molecule, and by
and multiple state structures of proteins, and also its importance for medical diagnostics using DNA
due to the many different water structures that are biochip arrays. More recently this interest has been
possible. stirred by potential applications of DNA in (bio-
Finally kinetic effects are of primary impor- mimetic) materials science and in molecular elec-
tance; the final structure of an adsorbed single tronics. Generally, much less has been done
protein, or protein adlayer, is almost always a regarding basic adsorption studies of nucleic acids
long-lived metastable state, rather than the lowest and DNA compared to amino acid, peptide and
energy state. In other words there is usually a protein adsorption, except in the sub-area related
sufficiently large activation barrier towards for- to DNA biochips. As a rule direct adsorption of
mation of the lowest energy states, so that further DNA on such surfaces is of less interest than ad-
conformational changes are prevented. A common sorption on pre-adsorbed spacer layers. The rea-
experimental manifestation of this is that the final son is that the surface perturbation from, say a
structure of a protein adlayer often depends on the metal, oxide or semiconductor surface, turns out
rate by which it is formed; at low bulk concen- or is suspected to be too strong to preserve the full
tration of proteins the adsorption is slow and there function of the DNA molecule. For example, it is
is then longer time available for conformational very likely that the dangling bonds of a non-
changes to occur, in comparison with high bulk hybridized DNA segment will bind to the surface
concentration and high adsorption rates, when and then be useless for detection of matching
neighboring molecules may sterically prevent slow segments. The maintenance of the full recognition
conformational changes. The issue of protein function of e.g. a single strand DNA segment is
conformation and denaturation at surfaces con- crucial for diagnostic applications as on biochips.
nects protein adsorption to the broad field of Therefore various spacer layer and linker molecule
protein folding–unfolding in the bulk phase (see strategies are currently developed to achieve both
e.g. Refs. [38,43] and references therein). localization of the diagnostic DNA segments at
A quantitative first principle description of different spots on a surface array, and preserved
protein bonding at surfaces lies far into the future. functionality.
It will be preceded by corresponding descriptions The molecular electronics potential of DNA is
of adsorbed amino acids and di- and tri-peptides currently a hot topic. Specifically there is intense
(and the influence of water on such bonding). It research addressing the question whether the DNA
will also require a quantitative first principles total molecule is a 1D conductor or not, and in that
energy description of proteins in the bulk phase, case, what the conductivity mechanisms are. Re-
including the description of dynamics and multiple cent literature reports both high conductivity and
states. Spectroscopic techniques such as non-linear insulating properties. Although some reports may
optical techniques and other laser based techniques be plagued by experimental difficulties there is
(SFG, SHG,. . .), synchrotron based spectroscop- another aspect; the question of conductivity or not
ies, neutron scattering, NMR, and scanning probe must be more specific, and discriminate between
B. Kasemo / Surface Science 500 (2002) 656–677 671

different types of molecules, and take into account tures that minimize the water interaction of the
if the measurement is made with or without pres- hydrophobic tails with surrounding water, and
ence of the water shell of DNA etc. Microscopic maximize the exposure of the hydrophilic end
(e.g. by SPMs) and spectroscopic characterization groups towards the water. This can result in many
of the actual DNA configurations, preferably in different self-assembled structures of which so-
the liquid phase, is thus vital for progress in this called unilamellar or bilayer membranes are the
field. most interesting in the present context. They typ-
ically consist of a double layer of phospholipid
3.1.5. Biomembranes molecules, with opposite orientations of the two
Biomembranes are and will be a future ‘‘hot’’ layers i.e. with two consecutive layers of parallel
topic for BioSS and many other sub-disciplines, hydrophobic tails in the interior, terminated on
due to their enormous importance as functional both sides by the hydrophilic end groups.
units in living cells, and because of their potential A specific structure occurs when the bilayer
role in many applications [46–53]. Biomembranes forms a closed spherical shell, with water both on
(Fig. 8 lower picture, Figs. 9 and 10) are self- the inside and the outside. This structure is named
organized structures of amphiphilic lipid mole- a vesicle or liposome, an entity that is a first
cules, the latter consisting of a hydrophobic crude approximation of a closed cell membrane,
(weakly interacting with water) lipid tail (linear however, lacking the functional units of real cell
hydrocarbon chain) and a hydrophilic (strongly membranes such as membrane-bound proteins,
interacting with water) terminal group such as ion channels etc. One could refer to them as a
a phosphate group (phospholipid molecules). In primitive, ‘‘empty cell’’ that can be filled by both
water solutions the thermodynamics drive these membrane-bound functional units (membrane
amphiphilic molecules to (self)assemble into struc- proteins) and intracellular components, in order to

Fig. 9. Formation of SPB from vesicles (schematic; not proven by experiments).


672 B. Kasemo / Surface Science 500 (2002) 656–677

Fig. 10. A sequence of preparation steps that have been experimentally implemented [50] to build up a biofunctional surface for e.g.
cell culturing or biosensing. The first step consists of formation of a biotin doped, phospholipid bilayer from vesicles, on top of which
(2nd step) a 2D crystalline protein layer (streptavidin) is adsorbed. The third step is adsorption of DNA or PNA molecules, which are
biotin coupled to the remaining free attachment sites on the streptavidin molecules. This platform can then be used e.g. for dis-
criminative recognition of perfectly and non-perfectly matching DNA segments.

successively build up more cell-like structures. brane proteins or protein or DNA layers on top of
Another common structure is the planar bilayer the membrane, can also be formed [48–53]. Pro-
membrane, which is called a supported (bilayer) teins are e.g. incorporated into the membrane by
membrane when deposited on a suitable surface. placing them inside the vesicles/liposome mem-
Such supported membranes are of extensive in- brane prior to surface deposition. The proteins
terest both as key elements of biosensor devices, incorporated in the membrane can then in turn act
and as mimics of ‘‘empty’’ or real cell membranes, as a binding site for other proteins on top of the
the latter after they have been ‘‘doped’’ by incor- membrane, e.g. through antibody–antigen binding.
porating membrane proteins etc. When the bi- A beautiful example reported by Reviakine and
layer is made up of phospholipids, it is called a Brisson is the 2D protein crystals on top of a biotin
supported phospholipid bilayer or biomembrane doped bilayer imaged by AFM [49]. By suitable
(SPB). Such bilayer membranes are also of interest linker chemistry one can continue to add additional
as functional coatings on medical implants, cell layers, to achieve a specific desired function. One
culture surfaces, and tissue engineering scaffolds. such sequence of preparation steps executed by
Supported bialyer membranes can be formed on H€ €k et al. [50] is shown in Fig. 11, beginning with
oo
a surface starting from vesicles/liposomes (see Refs. a supported membrane containing biotin, followed
[46–50], and references therein). Fig. 10 shows in by binding a 2D (crystal) of streptavidin on top,
schematic form how such supported bilayers may and completed by binding another biotin layer
form. Supported bilayers with incorporated mem- linked to a single strand DNA or PNA segment in
B. Kasemo / Surface Science 500 (2002) 656–677 673

the outermost layer. This structure can then be 3.1.7. Tissue


used to detect recognition events between the single Tissues at surfaces, in vitro and in vivo, have
strand DNA segment and its matching counter- been demonstrated to be sensitive to the adjacent
part, forming a double helix. For sensor applica- surfaces, but the mechanisms are understandably
tions it is important to find methods to pattern cells even less known than in the case of single cells
at surfaces. Such patterning has recently been and cell monocultures. Current research on tissue-
demonstrated by several groups, see e.g. Ref. [53] material interfaces in vivo follows several parallel
and references therein. lines. One major direction is chemical modifica-
tion of surfaces to enhance their biocompatibility
3.1.6. Cells and/or to provide controlled release of substances
Cells at surfaces is a fundamental ingredient that promote the healing and sustained function of
both with regard to the cultivation of cell cultures implants. Another one focuses on topographic
in vitro, tissue engineering, and medical implants. modification of surfaces to promote desired cell–
In the future it may also be employed in cell-based surface responses, and to some extent also pro-
biosensors and bioelectronics. The role of the tein–surface interaction. In some studies these
surface chemistry and topography for the evolu- surface modifications, i.e. chemical and topo-
tion and function of single cells and cell assemblies graphic, are combined to achieve optimal perfor-
has been demonstrated in many experiments [54– mance of the surface. Additional functions of the
63] but is still poorly understood. The recent rapid surface may be to e.g. act as a reservoir that can
development in the area of so-called stem cells is release certain desired substances, such as growth
particularly interesting in this context. Stem cells hormones, in a time-programmed manner.
are cells that have not reached a high degree of In the rapidly expanding tissue engineering field
specialization, but can ‘‘choose’’ to differentiate in the questions and challenges are to some extent
different specialized directions depending on the similar as for the in vivo situation with medical
local environment. Depending on the latter stimuli implants. There are however also entirely new
the cell can thus differentiate into different func- challenges connected with the fact that tissue en-
tions. It has also been shown that some stimuli can gineering demands that a given cell or tissue culture
make a cell ‘‘go backwards’’ from a specialized to shall evolve into a desired tissue without the sup-
less specialized state. port from the living organism that is the host for
The chemistry and topography of surfaces on in vivo implants. Thus tissue engineering demands
which cells grow/divide are part of the total envi- scaffolds that cooperatively with other factors
ronment for the cells and can potentially influence (nutrients, extracellular chemicals, force fields,
their differentiation, i.e. they are part of the total temperature, . . .) steer the tissue culture in the re-
cell–cell signaling/communication system. One key quired direction. In some sense one can say that
research area for the future is therefore how tai- the bioreactor must do for the tissue engineering
lored surfaces should be designed to act as stimuli scaffold þ cell culture, what the living organism
to guide cell differentiation, e.g. in tissue engi- does for the medical implant during the healing-in
neering, bioelectronics and cell-based sensors. For period. However, the total demand is much higher,
the surface scientist the chemical and topographic namely to make the orininal cell/tissue culture de-
patterning of surfaces, and the associated charac- velop into a functioning organ. This may require
terization is a major opportunity and challenge for special 3D architectures ranging from macroscopic
future research. Another is to use SPMs to probe dimensions to micrometer and nanometer dimen-
or stimulate specific cell functions. Yet another sions, with superimposed chemical patterns and
one is non-invasive, spatially resolved spectro- (bio)chemical signals, and probably with a time-
scopies and microscopies to study special fractions programmed degradation rate of the templates as
of living cells or cell membranes at high spatial the tissue matures. The research opportunities for
resolution (bioimaging) and sensitivity (single surface scientists in this area are similar to those for
molecule detection). cell–surface interactions (see above).
674 B. Kasemo / Surface Science 500 (2002) 656–677

3.1.8. Microorganisms terns derives from the very advanced recognition


Microorganisms at surfaces are important e.g. power of biological systems discussed earlier.
in marine biofouling and bacterial infections on Native material surfaces are very unsophisti-
implant surfaces or in tissue cultures. They are also cated compared to the biomolecular chemical þ
potentially of interest for growing simple organism topographic architectures they are intended to in-
cultures as model systems for a variety of biolog- teract with, and will in most cases at best cause a
ical experiments e.g. related to food production, mild negative response in the biological system. In
development and evaluation of pharmaceuticals, order to obtain a positive and selective response
and biomimetic energy production. Colonization the molecular architecture should in general match
of surfaces by e.g. bacteria (biofouling) is surface some recognition sites of biomolecules on the bi-
specific and can be influenced by surface chemis- ological side of the hybrid interface, so that it
try, surface topography, and surface visco-elastic- generates desired signal patterns to adjacent bio-
ity. The settling down of microorganisms on a molecules or cells or sensing devices. This type
surface is––as in the case of cell adhesion––almost of topographic þ chemical microarchitectures are
always preceded by protein adsorption. There is necessary both for applications and understand-
thus a causal relationship between protein ad- ing-oriented basic research of biointerfacial pro-
sorption and biofouling by microorganisms, in a cesses.
similar way as sketched earlier for cell adhesion on Consequently the future development of bio-
a surface. Also the basic research opportunities logically functional surfaces demands a very so-
and tools are similar as for cell culture growth and phisticated machinery for surface preparation and
tissue engineering, while the more applied research characterization, including dry and wet chemical
may require much more specific and specially adlayer depositions, and nano- and microfabrica-
adapted tools, e.g. to study ‘‘real’’ marine bio- tion.
fouling or bacterial colonization on medical im-
plants.
5. Methods

4. Surfaces Since this article is more concept and idea ori-


ented than method oriented this paragraph is de-
The material surfaces include almost all ma- liberately very brief. A general statement about
terial types; metals, ceramics, carbon materials, surface science methods and biointerfaces would
polymers and composite materials. Although be too trivial, since it would conclude that essen-
many native materials are currently used in prac- tially all methods of surface science come into play
tice, such as titanium for dental implants, stainless in some contexts.
steel for orthopedic implants, PTFE for blood Well-established examples on the analytical side
vessel replacements, silicones for internal drainage, are the use of established surface spectroscopies
PMMA for intraocular lenses, and so on, future (XPS, AES, SIMS,. . .) to characterize medical
more advanced materials and applications will implants after manufacturing and sterilization, but
often require the build up of sandwich type over- before clinical insertion, and also after varying
layers, with specific topographies and patterns, on periods in vivo (retrieved implants). Synchrotron
the native surfaces, to obtain the desired function. based spectroscopies will no doubt be important
This is actually already the case e.g. for intraocular tools both for spectroscopic characterization,
lenses made from PMMA and almost always the structure determination, and for imaging of bio-
case for advanced biosensors. In addition we know logical components on surfaces. With regard to in
that surface patterns, topographically and chemi- situ characterization of surface bound biomole-
cally, on different length scales generate different cules and larger entities, and especially for real-
responses of the biological system. This require- time recording of kinetic processes, the optical
ment of sophisticated surface structures and pat- techniques are of prime importance. This includes
B. Kasemo / Surface Science 500 (2002) 656–677 675

both linear and non-linear (laser) techniques, and probe techniques and time resolved laser spec-
all spectral regions from UV and down in wave- troscopies will be central tools in the development
length to the far IR. There is no doubt that scan- of BioSS.
ning probe techniques will continue to increase in
importance for studies of biomolecules at surfaces.
This includes both STM and AFM [64,65] and 6. A look into the future
spatially resolved optical techniques like SNOM.
Complementary techniques that provide unique When surface science started to take off as a
information in special cases, when sufficient sur- research field on its own, over thirty years ago,
face sensitivity is achieved are e.g. NMR, neutron little was known even about the simplest model
scattering and ESR. systems and the number and sophistication of the
Preparation by thin film deposition methods experimental and theoretical tools were very lim-
(PVD, CVD) and by glow discharge plasma ited compared to today. The electronic structure of
methods are common in R&D on hybrid surfaces was largely unknown, we did not know
biointerfaces. Wet chemical methods such as e.g. how low energy electrons were scattered by
self-assembled monolayers, colloidal chemistry, surface atoms, hampering the use of LEED as a
microemulsions, and micelles and vesicles come quantitative tool, the positions and orientation of
into play because they have the ability to produce CO on any surface were totally unknown, and we
interesting chemical and topographic patterns on were very far from any quantitative potential en-
surfaces. In addition they are usually fast and ergy surfaces for dissociation or reaction dynam-
often inexpensive. Synthetic polymer, protein and ics (even the 1D cuts were unknown). Total
oligonucleotide chemistry are increasingly impor- energies of chemical accuracy were impossible to
tant because they can produce ligands that can calculate.
selectively anchor desired polymer chains, pep- For simple systems the situation is today re-
tides, oligonucleitides or whole proteins to sur- versed; the total energies can be calculated, the
faces. electronic and atomic structures, and the lattice
The whole range of nano- and microfabrication dynamics of surfaces are known in great detail.
methods constitute important tools for the func- Potential energy surfaces can be calculated for
tional patterning of surfaces. They include electron dissociation and reaction of simple molecules, and
beam and photolithography and the soft lithog- the understanding is conceptually transparent even
raphies, like stamping and imprinting. for much more complex systems. Important fac-
Interesting to note are the special demands on tors have been the development of new experi-
methods for BioSS, set by the fact that one deals mental probes, advanced preparation of well
with very fragile matter, often requiring a liquid defined model systems, development of theoretical
environment for meaningful studies. This calls for methods and simulation schemes to describe both
new developments and/or to adaptations of ex- the studied model systems and how the experi-
perimental techniques to work non-invasively with mental probes (electrons, photons, ions, SPM tips
e.g. cells under liquid environments, or perform etc.) interact with surfaces, and a vast increase in
sophisticated freeze drying to retain the features to computational power. This development has al-
be studied by methods that require a vacuum lowed a broad and systematic, theoretical and
environment. For smaller entities like adsorbed experimental, exploration of model systems of
proteins and biomembranes, both the microscopic varying complexity, which successively paved the
inspection by SPMs and the spectroscopic char- way for a genuine, quantitative understanding to-
acterization under water are key issues. In order to day of the simpler model systems, and a semi-
address the important and challenging task of quantitative and/or conceptual understanding of
dynamics of e.g. proteins and supported mem- quite complex systems.
branes, femtosecond techniques are required, in This is the platform from which BioSS launches
the aqueous environment. Consequently scanning just now. We can foresee an exciting development
676 B. Kasemo / Surface Science 500 (2002) 656–677

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