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Nanotechnology and diagnosis
A.A. 2011-2012
Enhanced PCR
nAu reduces the formation of erroneous copies of
DNA in error-prone PCR.
M: markers,
nAu:
1: 0.6 nm
2: 0.0 nm
3: 0.2 nm
4: 0.4 nm
5. 0.8 nm
6: 1.0 nm
Enhanced staining of cell.
Thanks to their colloidal nature, the
nanomaterials interacts with the
biological systems.
Nanoparticles enters the cells and
differently accumulate at subcellular
level.
It is possible to target them to different
cell compartments/organelle by
selecting their physical-chemistry.
In the figure, differently sized QDs stain
extracellular space and subcellular
structures.
Enhanced “in vivo” imaging: QDs
A)Functionalized QDs
imaging of a rat with
muscle sarcoma.
B)The same as A: in this
case the QDs did not
have been
functionalized.
(Cai & Chen, Small, 2007, 3: 1840.)
Enhanced “in vivo” imaging: USPIO
A
B
MRI of brain lesion without (A) and with (B) contrasting with USPIO.
Biochemical monitoring
Higher glucose
Lower glucose
 Nanosensors are implanted
into the skin.
 They emit fluorescence
when irradiated by light.
 The fluorescence fades at
higher glucose concentrations,
because of the splitting of
fluorophore by competitive
binding.
The same procedure can be
applied to measure ions, pH,
osmolarity, etc.
Cash & Clark Trends Mol Med. 2010
The Lab-on-a-chip
Tseneva & Zhebrun, Pasteur Institute, St. Petersburg, ICPC Nanonet 3rd
Annual Workshop, St Petersburg, 24-25 May 2011
How does it work?
1. Analytes: blood, urine, saliva, gases in tissues, exhaled
air.
2. Integrated processes: sorption, chromatographic
separation, electrophoresis, thermocycling, (PCR), light
scattering, fluorescence.
Improved methods, implemented with nanotechnologies,
reduces sample volume and handling.
The lab-on-a-chip should save room, time and money.
Advantages: faster, cheaper and more accurate for multiple
analyses.
Main nanostructured
components.
Nanoporous Si-based membrane
A) membrane support
structure (A) with 5 µm
suspended (free standing)
membrane elements
(arrow).
B) Cross section of individual
element (arrow) showing
the porous nature of the
membrane
E,F) Pore shrinking protocol:
larger pores are oxidized
to be reduced by 50%
G) The pores at their final
limit (5 nm)
Johannessen et al. 2010. J Diabetes Sci Technol. 4(4):882-92.
Nanoporous Anodic Aluminium Oxide
(AAO) membrane
The figure shows a
nanporous membrane
of anodic aluminium
oxide (AAO).
Lateral (b) and top (c)
view.
The mean diameter of
nanopores is 200 nm.
The nanoporous
membranes are
implemented in devices
like those shown in the
following slides.
Ai et al. 2010 J Colloid Interface Sci. 350(2):465-70. doi: 10.1016/j.jcis.2010.07.024
Nanoporous membranes
Sensor (optical fiber)
Nanoporous membrane
Substrate
Signal
Complex bio-solution
Dialyzed target
proteins
Modified from: Chang et al. 2008. Nanotechnology. 19(36):365301.
doi:10.1088/0957-4484/19/36/365301
Nanochannels
Green arrow: constriction
of a nanochannel
(diameter = 130 nm)
The DNA strand enters into a nanochannel (1-3); its compression (4-6)
and relaxation (7-9) follow.
Reccius et al., 2008. Biophys J 95: 273–286
Separation of a single molecule of DNA into a silica-based
nanochannel.
+
The electrical field
is applied.
The DNA molecules
are drawn in the
loading zone,
-
The sample containing
DNA is inserted in a
chamber.
and assume an
elongated equilibrium
conformation in the
nanochannel
enter a nanochannel
Modified from: Liu et al., 2011 Electrophoresis, 32: 23: 3308-3318, DOI: 10.1002/elps.201100159.
Nanomixers
1. The fully assembled device
2. The 3D structure of the
mixer
3. The serpentine microfluidic
channel (50>300 nm)
4. The structures at the outlet
of the mixer
5. The structures in the middle
of the mixer
6. Cross-sectional view of a
structure similar to the ones
used in the mixers.
Jeon et al. 2005. Nano Letters 5(7): 1351-1356
The mixing process
b
The the serpentine with (a) and without (b) flow at 13.3 mm/sec.
Jeon et al. 2005. Nano Letters 5(7): 1351-1356
Nanopipette
1. Si-based, inner diameter: 100 nm
2. C-based with nAu coating
3. C-based, crushproof (a) and
injecting a cell (b)
1. Joe et al. 2006. J. Am Chem Soc 128(51): 16462-16463 .
2. Bau, Schrlau, Falls and Zinber, University of Pennsylvania School of Medicine.
3. Bhattacharyya et al. 2009 Adv. Mat. 21(40): 4039-4044 doi: 10.1002/adma.200900673
Nanosensor
A sensor based on nanotechnology (Si and metal oxide, STM 90 nm;
90 nm refers to the transistor gate length).
Johannessen et al. 2010. J Diabetes Sci Technol. 4(4):882-92.
What could the lab-in-a-chip do?
Cell separation
Rapid cancer detection
DNA analyses, PCR
Microtitration
Blood analyses
Microbiology platforms: growth,
identification and classification, antibiotic
sensitivity tests
 Capillary fluorimetric detection






Lymphocyte (WBC) isolation
1. Loading the blood sample
3. Labeling the WBC with fluo
2. WBC isolation, RBC elution
4. Washing the unbound dye,
reading fluo
Modified from: Zimina & Lucinin, 2011. J. Anal. Chem. 66(12): 1136–1158.
Rapid cancer detection
The hmw DNA (high molecular weigth), a marker for leukemia, is
captured from 20 l of untreated blood by the electrophoretic
circuit of a chip, activated with a fluorescent green dye. Wash it.
On the right (A) the result for an healthy subject, on the left (C)
that of a patient with leukemia.
(Heller et al., 2011. doi: 10.117/2.1201007.003153)
DNA analyses, PCR
1. Heater
2. SiC thermosensor
3. Radiator, with the thermosensor
shown in red
4. Separation channel and
reservoir for thermocycling.
5. Pattern of t°C cycles
1
2
3
4
The PCR on-a-chip
The
nanoporous
membrane
Modified from: Kim et al. 2010 Analyst. 135(9):2408-14
Continuous glucose measure on-achip.
Si membrane
Johannessen et al. 2010. J Diabetes Sci Technol. 4(4):882-92.
Testing antibiotic sensitivity
The picture….
and the scheme
Tsou et al. 2010. Biosens Bioelectron. 26(1):289-94.
The results
Partial
resistance
Resistance
Sensitivity
0
60
120
Tsou et al. 2010. Biosens Bioelectron. 26(1):289-94.
180 min
The goal: the microbiology lab
on-a-chip
1. Culture growth platform
2. Pre-filter
3. Sample
4. Image recognition
5. Laser detector (LIF)
6. Colorimeter
7. Microfluidic sorting element
8. CDD camera (low resol.)
9. AST microplatform
10.Pathogen growth
11.Bank of antibiotics
12.Lead
13.Substrate
14.Heater
15.Waste basin
16.Sorting way
Zimina, ICPC NANONET workshop, 24-25 May 2011, St. Petersburg
Another goal: the metabolic
lab-on-a-chip
Chang, 2004. Proc IEEE, 92(1): 154-173.
Another goal: the hematology and
clinical pathology lab on-a-chip
Electrophoresis
White cell count
Emocromo
White cell
histology
Heater
Proteins
pH
Sample well
Coagulation
Metabolites
Reaction centre
Kutuzov, St. Petersburg State Electrotechnical University ICPC Nanonet 3rd
Annual Workshop, St Petersburg, 24-25 May 2011