Updated on 03-07-2010

Research Projects Organization


XIAOHUA  WANG
 
     INVESTIGATION of the DYNAMICS of CONFINED FLUID-LIKE FILMS using
     SHEAR-FORCE/ACOUSTIC 
NEAR-FIELD MICROSCOPY (SANM)
     Implications to Nanotribology, protein folding/unfolding
  
I.  HANDS-ON TASKS:
 
     I.1. Using the commercial AFM:
  
           - Implementation in lithography-mode. (Document detailed procedure.)
              - Measurement of the AFM-cantilever spring constant (using the Park-AFM software, correlation with the
                 theoretically calculated values.)
              - Drive the Park-AFM externally (for obtaining images using an AFM probe as well as with a TF probe)
              - Implementation of a humidity-controlled small chamber (enclosing just the scanning stage.)
              - Implement Dip Pen Nanolithography (for triggering mechanical swelling of polymer films using electric fields.)
              References
              [1.1]  C. Maedler, S. Chada, X. Cui, M. Taylor, M. Yan, and A. La Rosa, “Creation of nanopatterns by local protonation of P4VP
                        via dip pen  nanolithography.” J. Appl. Phys. 104, 014311 (2008).
              [I.2]  C. Huang, G. Jiang, and R. Advincula, "Electrochemical Cross-Linking and Patterning of NanostructuredPolyelectrolyte-Carbazole
                       Precursor Ultrathin Films,"  Macromolecules  41, 4661-4670 (2008).

     I.2. Using the homemade microscope stages        
              - Approaching-retraction curves monitored by tunneling, acoustic, and tuning fork signals.
                 It involves the use of silicon-substrates and tapered-probes coated with hydrophobic and hydrophilic
                 polymer films.
              - Fabrication of probes of different geometries (sharp and chubby) and materials (tungsten and gold)
              - SEM images of the homemade probes (using the ISI-130 system available in the lab.)

II.  RESEARCH SCOPE:
      Investigation of the dynamics of confined mesoscopic fluid-like films [Ref II.1] at the nanometer scale
      using acousto/shear-force scanning microscopy
[Ref II.2]  
 
       II.1 TECHNOLOGICAL and SCIENTIFIC BACKGROUND   (Why is the thesis topic important, what type of
                problems  it aims to contribute in their solution, ...  etc. See Ref II.1)

               II.1.A  Mesoscopic fluid-like films
                             Surfaces are involved in the strikingly different dynamics that emerge from adsorbed mesoscopic fluids. Shear viscosity
                             is enhanced, viscoelastic relaxation times are prolonged, and fluid phase transitions to solid or liquid are seen to be induced
                             by confinement. Ref [A.1], [A.,3], [A.,4] But, what determines such behavior? What is the molecular organization of the adsorbed
                             mesoscopic liquid layer that leads to the observed differences in dynamics compared to the bulk? What are the dynamics of liquids
                             in intimate contact with a solid boundary?  Which aspects of the behavior of a mesoscopic liquid can be attributed to confinement
                             alone? These questions identify some of the unresolved issues associated with surface phenomena.  Additionally, there is evidence
                             that surfaces and interfaces modify the relaxation rate towards equilibrium of polymeric films when cooled to temperatures
                             below the glass transition temperature; understanding the glassy state and its associated phenomena are also central
                             challenges in condensed matter physics. Ref [A.5] This thesis aims at characterizing
surface, interfaces, and the dynamics of
                             confined mesoscopic fluids using the novel Shear-force/Ultrasonic technique.
                             References:
                             [A.1]  S. Granick, “Motions and Relaxations of Confined Liquids,” Science 253, 1374-1379 (1991).
                            
[A.2]  A. La Rosa, "An investigation of the dynamics of confined fluid-like films under shear-forces at the nanometer scale."  
                             [A.3]  M. Urbakh, J. Klafter, D. Gourdon, and J. Israelachvili, “The nonlinear nature of friction,” Nature 430, 525-528 (2004).
                             [II.4]  J. Klein and E. Kumacheva, "Confinement induced phase transition in simple liquids," Science 269, 816 (1995).
                             [A.5]  R. D. Priestley, C. J. Ellison, L. J. Broadbelt, and J. M. Torkelson; “Structural Relaxation of Polymer Glasses at Surfaces,
                                        Interfaces and in Between,” Science 309, 456 (2005)
                             [A.6]  S. Davy, M. Spajer, and D. Courjon, "Influence of the water on the shear foce damping in  near-field microscopy,"

                                        Appl. Phys. Lett. 73, 2594 (1998).
                             [A-7]  Maria-Pilar bernal, et al, "Theoretical and experimental study of the forces between SNOM probes and chemically treated
                                       AFM cantilevers," Proceeding of the IEEE 88,1460 (2000).
                             [A-8S. Biggs and P. Mulvaney, Measuring of forces between gold surfaces in water by_AFM, J. Chem. Phys. 100, 8501 (1994).
                                       
A long range attraction is seen, consistent with the large Hamaker constant for goldlwater/gold.  Within the Derjaguin
                                        approximation, the force F between the sphere of radius R and the plate at a separation D, due to van der Waals forces,
                                        is given by F/R= -A(D)/6D*D, where A(D) is the Hamaker function at a separation D, obtained from Lifschitz theory.         
             II.1.B  Basics of Intermolecular forces 
                             -  Summarize Chapters 9, 10, and particularly 15, of Ref [B.1].
                                Highlight the concepts of  interfacial energy,  surface tension adhesion, cohesion, ...
                                Include concepts contrasting intermolecular and intersurface forces
                             -  Adhesion between two rigid (incompressible) macroscopic spheres
                                Adhesion  between spheres that deform elastically: the Hertz and JKR models (the latter takes into account adhesion forces).

 

                             References:

                             [B.1] Jacob N. Israelachvili, Jacob N., " Intermolecular and surface forces," Available at PSU Library: QD461 .I87 1991.
 
                             [B.2] C. Rotsch, K. Jacobson, and M. Radmacher, “Dimensional and mechanical dynamics of active and stable edges in motile
                                      fibroblasts investigated by using force microscopy,” Proc. Natl. Acad. Sci. USA 96, 921 (1999).

                                      (It uses Hertzian model to calculate the Young modulus E of fibroblasts. This article  may be useful to learn how to apply the
                                      Hertz model to obtain Young modulus values  from approaching and retraction curves data.)

              
II.1.C  The Hydrophobic and Hydrophilic Interactions
                             What are the hydrophobic and hydrophilic interactions?
                             - Make a review summary of Chapter 8 of Israelachvili's book Ref [C.1]             
                             - In Israelachvili's book Ref [C.1] check section 15.5 and, in  particular, the 3rd paragraph on page 329, where the author
                                highlights the potential 
different molecular rearrangements during the approaching and retraction. The latter may account for the
                                cases where the
Hertz and JKR models (the latter takes into account adhesion forces) fail.
                             - In Ref [C. 1] check section 13.6 that makes reference to the "attractive hydrophobic forces".
 
                             Relevance of the hydrophobic and hydrophilic interactions
                             Water at interfaces is fundamental to the understanding of wetting, foldin/unfolding of macromolecules, tribology, chemical
                             reactivity and dynamics.

                             Hydrophobic and hydrophilic interactions on amorphous and crystalline substrates
                             Amorphous polymer samples
                                      In this thesis we are preparing polymer samples with well controlled hydrophobic and hydrophilic properties. [ Xiaohua,
                                      Dr. Yan told me once that her lab can even control the degree of hydrophobicity or hydrophilicity. But I am not so sure.
                                      Check with Suji if this is correct. If so, it would be a good way to verify whether samples of  higher hydrophobicity or
                                      hydrophilicity produce more or less sound.)
                             Crystalline hydrophobic graphite
                                      For comparison purposes, this thesis will include approaching/retraction curves using commercially available hydrophobic
                                      crystalline graphite samples
                             References:
                             [C.1] Jacob N. Israelachvili, Jacob N., " Intermolecular and surface forces," Available at PSU Library: QD461 .I87 1991. 
                             [C.2] Ding-Shyue Yang and Ahmed H. Zewail, “Ordered water structure at hydrophobic graphite interfaces observed by 4D,
                                      ultrafast electron crystallography,” PNAS
106, 4122 (2009).

               II.1.D  Quantum phenomena 
                             Include a perspective of  future studies to your thesis theme: Review the Casimir-Lifshitz force subject.
                             References: 
                             [D.1]  J. N. Munday, F. Capasso, and V. A. Parsegian,  "Measured long-range repulsive Casimir–Lifshitz forces," Nature 457,
                                       170 (2009).
                             [D.2]  Katharine Sanderson, "Quantum force gets repulsive," Nature 457, 156 ( 2007).
                             [D.3]  J. N. Munday, F. Capasso, V. Adrian Parsegian, and S. M.  Bezrukov, "Measurements of the Casimir-Lifshitz force in fluids:
                                      The effect of electrostatic forces, and Debye screening," Physical Review A 78, 032109 (2008).
                             [D.4]  Philip Ball, "Fundamental physics feel the force,"  Nature 447, 772 ( 2007).

       II.2 CHARACTERIZATION TOOLS
                To study such complex surface phenomena, this thesis will exploit the versatile capabilities of the Near-field Ultrasonic/Shear-force
                Scanning Microscope (NSUM) [Ref. II.10], a novel technique able to concurrently and independently monitor the effects that fluid-mediated
                interactions exert on both the microscope’s probe and the sample to be analyzed.            
                Provide a background of other techniques also being used to study mesoscopic films, like for example, the Shear-force apparatus

                References:
                [II.10]  A. H. La Rosa, R. Nordstrom, X. Cui, J. McCollum,"The Ultrasonic/shear-force microscope: Integrating ultrasonic sensing into a
                           near-field scanning optical microscopemicroscope," Rev. Sci. Instrum. 76, 093707 (2005).



RODOLFO FERNANDEZ

     DEVELOPMENT  of  FAR- and NEAR-FIELD OPTO/ACOUSTO  NANOIMAGING  TOOLS
     for BIO-MATERIALS CHARACTERIZATION at the MOLECULAR LEVEL

 
     Morphology, spectroscopy and si
ngle-molecule optical nano-localization

I. HANDS-ON TASKS

    I.1  Developing acousto/electronic position-control feedback for scanning probe microscopy
             I.1A Implementation of a (homemade) feedback electronic system for controlling the probe-
                     sample distance in near-field ultrasonic/shear-force scanning microscopes (NUSM).
                      - Integrating a linear scaner z-stage (Mad City Lab, MCL) with a xy-scanning stage (PiezoJena).
                        Evaluation of the MCL-stage's time response.
                      - Modification of the LabView-based programming feedback circuit design  and corresponding
                        customization of a FPGA card for automated control of the probe-sample distance.
                      - Optimization of a 16-bit resolution ADC for driving a 65-micron travel-range stage at nanometer steps motion
                         (Strategy: automated re-calibration, while scanning, of the actual z- travel range (1 micron) in order to improve the
                         ADC's resolution.)

                      - Implementing in situ automated adjustment of the proportional and integral gains during the scanning process.
 

             I.1B Integrating frequency-modulation feedback control  (Nanonis) into the NUSM
            
          - To build custom-made pre-amplifiers for proper conditioning of the signals.
                      - Acquisition of approaching/retraction curves while updating the z-dependent resonance frequency (where z is the probe-sample
                        distance.) Monitoring of the amplitude variation should provide
information of the damping force component.
                      - Acquisition of approaching/retraction curves with the sample immersed in water environment.
 
             I.1C Development of an  "Acoustic-based Electronic Feedback System" for probe-sample distance
                    control in tuning-fork based scanning probe microscopes.
                      - To exploit acoustic resonances of the microscope stage for sensitively monitoring the mechanical vibrations of the tuning fork. Look
                         for optimized detection schemes for monitoring the nanometer-size variations of the TF amplitude upon its interaction with the
                         interrogated  sample.
             References
             [I.1.1]  J. Jersch, T. Maletzky, and H. Fuchs, “Interface circuits for quartz crystal sensors in scanning probe microscopy applications,”
                          Rev. Sci. Inst 77, 083701 (2006).
   Comments: It describes PLL control feedback of 1Mhz tuning forks.
             [I.1.2]  U. Rabe and W. Arnold, " Acoustic microscopy by atomic force microscopy, "Appl. Phys. Lett. 64, 1493 (1994)
                          Comment: It uses knife edge detection for optical detection of acoustic waves.
             [I.1.3]    R. L. Whitman and A. Korpel, "Probing of Acoustic Surface Perturbations by Coherent Light," Appl. Opt. 8, 1567 (1969).
                          Comment: It describes the use of knife edge detection for optical detection of acoustic waves.

    I.2  Developing FAR- and NEAR-field Optical Nanoscopy
          I.2A Fabrication of NSOM probes
                      - Fabrication of tapered glass-fiber probes by chemical (HF) etching.
 
          I.2B Far-field optical tracking of individual fluorophores with nanometer lateral resolution
                      - Solid-graphic design for modifying of the IX-71 Epi-Fluorescence Inverted Microscope.
                         (Design of a sturdy sample-stage for alternatively holding a near-field optics scanning stage.
                         Design of a lifted microscope's sample-stage for future laser trapping experiments.)  
          I.2C Far-field Nanoscopy: Stimulated Emission depletion (STED)
                      - To reduce the effective size of a diffraction-limited light spot (light-beam-1) with a pico-second doughnut-shape (beam-2).
                      - Select the proper flurophores for the techniques testing purposes.
                      - Undertake test with samples under aqueous conditions
 
          I.2D Implementation of imaging/spectroscopy capabilities for single molecule studies
                      - STED imaging at the interior of cells
                      - Integrate the EM CCD camera  (PhotonMax 512B) of single-photon sensitivity and the spectrometer XXX with the IX-71 inverted
                        microscope.

 
             References: 
             [I.2.1]   R. E. Thompson, D. R. Larson, and W. W. Webb, "Precise Nanometer Localization Analysis for individual Fluorescent Probes ,"
                          Biophysical Journal 82, 2775 (2002).
                          The image-size of an object is limited by diffraction. However, the center of the object can be determined arbitrarily precisely,
                          given a sufficient number of photons (N) in the spot. Two important source of noise affect this method: a) the shot noise of the
                          photons in the image spot, and b) the background noise created  by out-of-focus fluorescence, charge coupled device (CCD) 
                          readout noise, dark current, and other factors. 
             [I.2.2]   V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, Stefan W. Hell, “ Video-Rate Far-Field Optical Nanoscopy
                          Dissects Synaptic Vesicle Movement,” Science 320, 246 (2008).
             [I.2.3]   S. W. Hell, "Far-field Optical Nanoscopy," Science 316, 1153 (2007).
             [I.2.4]  Yildiz et al, "Myosin walks hand-over-hand, Single fluorophore imaging with 1.5 nm localization" Science 300, 2061 (2003).
             [I.2.5]  Yildiz et al, "Kinesin walks hand-over-hand ," Science 303, 676 (2004).
             [I.2.6]   D. A. Fletcher, and  R. D. Mullins, “Cell mechanics and the cytoskeleton,” Nature 463, 485 (2010).
                          Comments: Article displays neurons and microtubulus and actin filaments.
        
II. RESEARCH SCOPE
      DEVELOPMENT  of  FAR- and NEAR-FILED OPTO/ACOUSTO  NANOIMAGING  TOOLS
      for BIO-MATERIALS CHARACTERIZATION at the MOLECULAR LEVEL

 
      Morphology, spectroscopy and single-molecule optical nano-localization
       II.1 TECHNOLOGICAL and SCIENTIFIC BACKGROUND   (Why is the thesis topic important, what type of
                problems  it aims to contribute in their solution, etc. )

                II.1.A Current developments of nano-optical techniques (See Section II of Ref [II.1]. )
                             Conventional Optical microscopy would be the preferred tool for characterizing biological dynamic
                             events with nanometer spatial resolution given its simple use, relatively low cost, and, quite important, non-invasive character.
                             Unfortunately, diffraction effects prevent conventional  optical microscopy from providing spatial lateral resolution better than l/2
                             (where l ~ 500 nm is the wavelength of the radiation used) as  enunciated by Ernst Abbe in 1873.  In what follows we highlight how
                             new technologies are avolving to surpass
the DIFFRACTION BARRIER.
                                     
                             PIN-HOLE. In 1928, Synge conjectured that the diffraction-limited lateral resolution was not fundamental, but rather an inherent
                             constraints resulting from the (unavoidable at that time) use of lenses, which have a working distance (WD) greater than a few 
                             wavelengths (WD > l meansfar-field.”). Synge argued that IF a metallic pin-hole of sub-wavelength diameter were i) used as a photon
                             collector, ii) brought into the  proximity of the sample (WD <  l; that is “near-field”), and iii) laterally scanned along the sample’s surface,
                             THEN a lateral resolution equal  to the pin-hole aperture-size (i.e. sub-wavelength) could, in principle, be achieved.
 
                             NSOM. Based on an evolution of Synge's ideas, Near-field Scanning Optical Microscopy (NSOM), became the first optical
                             technique to overcome the diffraction barrier, as demonstrated in 1984. NSOM, however, has its own inherent limitations, one of them
                             being the typically low-level signal available for analysis. The smaller the aperture (aiming for a finer resolution), the more stringent the
                             requirements on the NSOM’s  detection system. NSOM beats the diffraction limited resolution by imposing the requirement to place
                             the excitation source very close to the sample. In many cases that is not possible to do (imaging inside living cells, for example.) Optical
                             imaging with subwavelength resolution remained a challenge.
 
                             STED. In 1994 J. Wichmann and Hell outlined a concept to eliminate the resolution-limiting effect of diffraction without eliminating
                             diffraction itself. It requires i) coating the sample with fluorophores, ii) excite the region of interest with a diffraction-limited light beam
                             (l1) iii) simultaneously applying a doughnut shape beam (l2), concentric with the excitation beam,  to quench the fluorophores  located on
                             the periphery. The net effect  is an excitation region of smaller dimension than l1.
 

                             FAR-FIELD FLUORESCENCE NANOSCOPY. A plethora of far field-based alternative solutions for attaining sub-wavelength
                             optical resolution have appeared lately (FIONA, RESOLFT, PALM), altogether referred to as Far-field Fluorescence Nanoscopy [II.2].
                            
They exploit basically the following principles to bypass Abbe’s barrier:
                                        
(1) Diffraction does not prevent finding out the coordinates of a point-source with arbitrary

                                               precision if there is no other similar point-source within a l/2 distance.
                                       
or
                                       
(2) Localization of individual fluorophores positioned within a distance much smaller than l/2
                                               can be  achieved by sequentially imaging them based on either different
                                               spectral response or selectively activating their “ON” and “OFF” states).

 

                             Principle (1 ) exploits the fact that when particles are widely spaced from each other (separation distance greater than the wavelength),
                             their diffraction-limited images may superimpose but the position of their corresponding centroids (spot of maximun intensity) can be
                             estimated with nanometer precision by fitting the experimental data (light intensity) to Gaussian functions. Hence, a direct correlation can
                             be made between the light point source and the center of its diffraction limitted image. Nanometer-sized step motion of the centroid have
                             been obtained with a CCD camera of sufficient dynamic range (i.e able to dictiguish fine difference in intensity levels.) The fitting method
                             has successfully been used to track the  motion of cargo-carrying myosin  with 2 nm precision, helping to figure out that they undergo
                             hand-over-hand motion.

                             This thesis aims at the development of a new Integrated Near- and Far- Field Optics (INFO) method for overcoming the
                             diffraction-limited lateral-resolution that hampers conventional microscopy is proposed. It envisions the use of a) a metal-coated
                             near-field probe, to optically isolate, one at a time, sparse fluorophore markers within a region of sub-wavelength dimension; and
                             b) a far-field localization method for locating the selected fluorophore marker’s position with nanometer precision. For its implementation,
                             INFO turns into an advantage the non-radiative energy transfer that fluorophores experience when placed in the vicinity of a metal wall.
                             Thus, fluorophores closer to the near-field probe’s rim (see figure below) become dimmer, which, by contrast, helps to localize more
                             clearly the fluorophore underneath the probe’s aperture that is less affected by the metal. A sequential repetition of the process a) and
                             b), in synchronization with the lateral scanning of the near-field probe, allows the localization of sparse fluorophores within a region of
                             sub-wavelength size hence overcoming the diffraction barrier. We will investigate the flexibility of implementing INFO with low and high
                             power laser system, as well as standard fluorophores (rhodamine), quantum dots, and photo-switchable fluorophores.


                             References:
                             [II.1] A. La Rosa et al,  "Integrated Near- and Far-Field Optics (INFO)"          
                                      Alternative method for overcoming the diffraction-limited  lateral-resolution that hampers conventional microscopy.
                             [II.2] S. W. Hell, "Far-field Optical Nanoscopy," Science 316, 1153 (2007).
                             [II.3] NALMS microscopy. Nanometer accuracy is demonstrated for two to five single molecules within a diffraction-limited area.
                                      X. Qu, D. Wu, L. Mets, and N. F. Scherer,"Nanometer-localized multiple single-molecule  fluorescence microscopy," PNAS
                                      101, 11298  (2004).
                             [II.4] PALMS MICROSCOPY Samuel T. Hess, Thanu P.K. Girirajan,  Michael D. Mason, "Ultra-High Resolution Imaging
                                      by Fluorescence Photoactivation Localization Microscopy," Biophysical Journal  91, 4258 (2006).  

                                      Authors make an estimation of the the requirements for  imaging with either 80-nm or 20-nm localization precision

JOHN LEDESMA
        DEVELOPMENT  of  PROGRAMABLE OPTO/ELECTRONIC  DEVICES
        for BIOSENSING APPLICATIONS
I.  TASKS:
 
     I.1  Fabrication of probes for Near-Field Scanning Optical Microscopy
          
(Apertures of sub-wavelengths dimensions, for optical imaging with high lateral resolution.
 
           I.1A  Metal coating tapered-fiber probes using a vacuum thermal evaporator. (Rodolfo will train John in the use of the thermal evaporator.)
           I.1B  Modification of probe morphology (to create a metalic aperture at the probe's apex) using a  focused ion beam system. (Rodolfo will
                        train John in the use of the FIB.)
              I.1C  Coupling laser light into the NSOM probes
           Note1:  Be patient with these steps as the thermal evaporator and the FIB  are sometimes temperamental, and coupling light into a fiber
                           sometimes takes 10 minutes and sometime one hour..
           Note2:  The tapered glass-fiber probes will initially be provided by another  graduate student who is already well-trained in handling HF acid)
           Note3: This task is urgently needed at the lab. If we can have those probes yesterday it will be great !!!
           References: 
           -  Application of NSOM: A. La Rosa, Near-field characterization of semicondustor nanostructures and devices
                 
           [Ref] Fabrication of probes by chemical etching R. Stockle, C. Fokas,V. Deckert, and R. Zenobi, " High-quality
                
near-field optical  probes by tube etching,"  Appl.Phys. Lett. 75,  160 (1999).
                        "A method called tube etching for the fabrication of near-field optical probes is presented. Tip formation occurs inside
                         a cylindrical cavity formed by the polymer coating of an optical fiber which is not stripped
away prior to etching in
                         hydrofluoric acid. ... In the case of fiber with permeable jacket, the tip forms
by the same mechanism as in the case
                         of the impermeable polymer coating."

           Introduction: Focused Ion Beam Systems
           Focused Ion Beam Principle
           A. La Rosa, "Near-field Scanning Optical Microscopy."
 
     I.2 Opto/acousto characterization of (antibody/antigen, protein-carbohydrate) molecular interactions

           1.2A Identify the proper  molecules/samples  that can optimize their molecular interactions
                       (We are pursuing the development of acousto detection methods developed by our group.)
 
                       Ref:
Questions: on how to optimize the experimental set up for acousto sensing molecular interactions
                               Find in this file some preliminary questions addressing the potential samples to be used in this project.
   
         
I.2B Optical Tracking of Individual Particles with Nanometer Resolution (LABELED MOLECULES)
                       "The image-size of an object is limited by diffraction. However, the center of the object can be determined arbitrarily precisely,
                       given a sufficient number of photons (N) in the spot. Two important source of noise affect this method: a) the shot noise of the
                       photons in the image spot, and b) the background noise created  by out-of-focus fluorescence, charge coupled device (CCD) 
                       readout noise, dark current, and other factors."
 
                      Task iscludes
                      
- Hands-on work integrating the PhotonMax EMCCD camera  (of single photon sensitivity) into the IX-71 Epi-
                         fluorescence inverted optical microscope.
                      - Identify proper sample for facilitating the application of the tracking techniques Again, sample preparation is key for the success
                         of tracking single fluorophores in in-vitro samples.
                       - Data acquisition tracking the position of  single molecules.
 
                      References: 
                      [I.2B1]  R. E. Thompson, D. R. Larson, and W. W. Webb, "Precise Nanometer Localization Analysis for individual Fluorescent Probes ,"
                                 Biophysical Journal
82
, 2775 (2002).
                      [I.2B2]  Yildiz et al, "Myosin walks hand-over-hand, Single fluorophore imaging with 1.5 nm localization" Science 300, 2061 (2003).
                      [I.2.B3] Yildiz et al, "Kinesin walks hand-over-hand ," Science 303, 676 (2004).
                      [II.2B4] A. La Rosa et al,  "Integrated Near- and Far-Field Optics (INFO)"
 
          I.2C Spectral analysis of single molecules (UNLABELED MOLECULES)
                      Ultimately, our objective is to spectrally identify  single molecules using Raman spectroscopy. This responds to the current interest for
                      characterizing unlabeled molecules, as typical  fluorophores could  affect the natural  dynamics  of the interrogated molecule.
  

     I.3 Build programanble BIO/MEMS devices


MICHAEL HOPKINS
     NEAR-FIELD  PROBES FABRICATION
 

I.  TASKS:
 
     I.1  Fabrication of FIBER-GLASS PROBES for Near-Field Scanning Optical Microscopy
          
(Apertures of sub-wavelengths dimensions, for optical imaging with high lateral resolution.
 
           I.1A  Rodolfo will provide the tapered glass-fiber probes 
          
I.1B  Metal coating tapered-fiber probes using a vacuum thermal evaporator. (Rodolfo will train Mike  in the use of the thermal evaporator.)
           I.1C  Modification of probe morphology (to create a metalic aperture at the probe's apex) using a  focused ion beam system. (Rodolfo will
                       train Mike in the use of the FIB. Subsequently, Mike will train Joshua in the use of the FIB.)
              I.1D  Coupling laser light into the NSOM probes (Gangandeep).
              I.1D  Implementation of acousto-optic modulation setup for amplitude modulation of laser systems (Gandeep).
           Note1:  Be patient with these steps as the thermal evaporator and the FIB  are sometimes temperamental, and coupling light into a fiber
                          sometimes takes 10 minutes and sometime one hour..
           Note2: The tapered glass-fiber probes will initially be provided by another  graduate student who is already well-trained in handling HF acid)
          
           References: 
           -  Application of NSOM: A. La Rosa, Near-field characterization of semicondustor nanostructures and devices
                 
           [Ref] Fabrication of probes by chemical etching R. Stockle, C. Fokas,V. Deckert, and R. Zenobi, " High-quality
                
near-field optical  probes by tube tching,"  Appl.Phys. Lett. 75,  160 (1999).
                        "A method called tube etching for the fabrication of near-field optical probes is presented. Tip formation occurs inside
                         a cylindrical cavity formed by the polymer coating of an optical fiber which is not stripped
away prior to etching in
                         hydrofluoric acid. ... In the case of fiber with permeable jacket, the tip formes
by the same mechanism as in the case
                         of the impermeable polymer coating."

           Introduction: Focused Ion Beam Systems
           Focused Ion Beam Principle
           A. La Rosa, "Near-field Scanning Optical Microscopy."

     I.2  Desingn and fabrication of COAXIAL PROBES for Near-Field Scanning Optical Microscopy
           I.1A   Fabication of tapered metallic probes (tungsten, gold) by etching processes  (Joshua)
           I.1A  Fabication of tapered probes out of commercially available glass pipettes using a commercially available pipette puller.
           I.1C  Fabrication of coaxial probes (Mike, Joshua).
           I.1D  Teraherta characterization of biomolecules (Gagandeep).

       References: 
           -  A. La Rosa, "Combined Terahertz/visible Near-field Optical Microscopy."
                  It includes different type of probes being pursued at Dr. La Rosa's Lab.

     I.3  Molecular layering in confined mesoscpic films
          [Ref-1]   M. Antognozzi, A. D. Humphris, and M. J. Miles, "Observatin of mlecular layering in a confined water film and study of the l
                           ayers viscoelastic properties. Appl. Phys. Lett. 78, 300 (2001).



KEITH PARKER

        1) Design and construction of high-sensitivity (108 gain), dual-stage (for increase bandwidth)  pre-amplifier.

            Focused effort on building a replica of existent design that includes a dual FETs (a design already proven
            to work well.)

                        Reference: R. D.Grober,  J. Acimovic, J. Schuck, D. Hessman, P. J. Kindlemann, J. Hespanha, A. S. Morse, K. Karrai,
                                           I.Tiemann, and S. Manus, “Fundamental limits to force detection using quartz tuning forks ,” Rev. Scientific. Instrum.
                                           71, 2776 (2000).

                    Comments: Our objective is to achieve their sensitivity detection levels
   
         2) Design and construction of a frequency modulation electronics circuit, for monitoring approaching-
             retraction curves along the z-direction. 
(No lateral xy scanning involved here.)                 
             Focused effort in building a replica of existent designs based on Edwards's and/or Kobayashi's papers.

                        Reference:  H. Edwards, L. Taylor, and W. Duncan, "Fast, high-resolution atomic force microscopy using a quartz tuning fork
                                             as actuator and sensor,"  J. Appl. Phys. 82, 980 (1997
).
                        Reference:  K. Kobayashi, H. Yamada, H. Itoh, T. Horiuchi, and K. Matsushige, "Analog frequency modulation detector for
                                              dynamic force microscopy,"
Rev. Scientific. Instrum. 72, 4383 (2001).

         3) Use the previous designs as a basis for a design based on more updated electronic components.



PROTEINS FOLDING/UNFOLDING


                      References:
                      -  Chritoph Kowitz,
The Entropic Force   (April-2008)
                      -   J. S. Graham, "
Mechanical Properties of Complex Biological Systems using AFM-Based Force Spectroscopy,"
                           Ph.D. Thesis (2005)