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LABS AT NIBIB
Nanoinstrumentation and Force Spectroscopy
The LCIMB’s Nanoinstrumentation and Force Spectroscopy (NFS) Section develops specialized instrumentations and their applications at macromolecular, cellular and tissue level in areas of biomedical research and medicine. NFS scientists collaborate closely with other intramural and extramural investigators to provide innovative approaches through biophysical modeling, mathematical analysis, and custom instrumentation primarily for nanoscale characterizations. Our current focus includes the development and applications of high-resolution and high-speed atomic force microscopy (AFM) for force spectroscopy and nanometric bioimaging of biological and soft materials. We also develop related technologies such as laser and optical technologies for spectroscopic analysis of biochemical reaction kinetics, multimodal instrumentations, and broader biomedical characterizations.
- Development of AFM Nanotechnologies
NFS Section shares a core biological atomic force microscopy (AFM) facility at NIBIB and extends the functionality of several commercial and developing AFM platforms toward higher speed, higher resolution, and multimodal measurements to yield better insights on biomolecules, their complexes, multifunctional theranostics, cells, and tissues. One example is an integration of optical spectroscopes with an AFM platform for tip-enhanced Raman scattering (TERS) and other multimodal characterizations. In another example, we have started an international collaboration dedicated to development and biomedical applications of a next-generation high-speed AFM. Over the years, our staff has had extensive experience improving such instrumentations to investigate a broad range of biomedical samples and systems.
- Biomedical studies via AFM and force spectroscopy
NFS Section continues to develop our biophysical measurement systems based on AFM imaging and single molecule force spectroscopy (SMFS), quartz crystal microbalance-dissipation (QCM-D), and other technologies, and to apply these technologies to a number of biomedical and biological investigations in collaboration with NIH intramural and extramural researchers. Major collaborations with notable results include: (A) Macromolecular structure and nanomechanical properties of multiple malaria vaccine candidates in collaboration with Dr. David Narum (Laboratory of Malaria Immunology and Vaccinology, NIAID, NIH) and other co-investigators. These protein antigen constructs are being produced via recombinant-protein biotechnology, purified, and characterized in a manner suitable for human trials and scale-up productions. We have focused on using AFM and QCM-D to understand the structural properties of these developing vaccines toward enhanced immunological response and eventual eradication of malaria. (B) Multifunctional nanomedicine probes with Dr. Shawn Chen (LOMIN, NIBIB), Dr. Ashwin Bhirde (LOMIN, NIBIB), Dr. Peng Huang (LOMIN, NIBIB), and many co-investigators. New results on several multi-functional nano-drug, photothermal therapy (PPT), siRNA delivery and imaging systems have been published and developed further toward cellular and medical applications. (C) Protein domain structure and interactions clathrin and clathrin assemblies in receptor-mediated endocytosis and intracellular trafficking in collaboration with Dr. Ralph Nossal (NICHD, NIH), Prof. Eileen Lafer (Univ.TexasHealthSciencesCenter, San Antonio), and coworkers. (D) A number of other intramural and extramural collaborations involving Bio-AFM, QCM-D, and related nanotechnologies for macromolecular and cellular studies.
- AFM and spectral analysis of biochemical reaction kinetics
Bacteriorhodopsin (BR) is a bacterial, membrane-bound proton pump that is a prototype for mammalian cytochrome oxidase of respiratory chains. It is a well characterized protein with spectral signatures in both the visible and infra-red (IR) regions of the electromagnetic spectrum. Previously in collaboration with NHLBI/NIH and NIST investigators, we produced a multi-channel visible spectrum analyzer that had a time resolution of 5 microseconds, which enabled characterization of changes to the spectral signature that occurred following initiation of the BR photocycle by an actinic laser photolyzing pulse. Using this new instrumentation and a linear algebraic approach following a singular value decomposition of a matrix of the raw data, we have isolated the absolute visible and infrared spectra of all of the photocycle intermediates. Recently, our team has developed a new system to perform the same analyses and compare the behavior BR in its native purple membrane environment (PM) with that in isolated crystals. This new mutli-channel analyzer, which offers sub-microsecond time resolution, incorporates a fiber optically coupled spectrograph comprising a frame-shift ccd camera, and a time-gated image intensifier, with an IR/visible microscope that has its own IR detector. The sample, whether PM or a crystal is contained in a defined 50 micron by 50 micron space. The ultimate goal is two-fold. First, to validate the biological relevance of BR crystals; and secondly to use the same linear algebra deconvolution approach to isolate the X-ray diffraction maps and real structures of the photocycle intermediates. This should help in understanding how protein conformational changes during the pumping of protons across the membrane form the electrochemical potential that drives ATP synthesis. In a parallel study, we are combining optical spectroscopy with AFM imaging to better understand conformational changes and polymerization of amyloid beta protein in Alzheimer's disease (AD).
Biological AFM capabilities
Combining high-resolution AFM under physiological conditions with sensitive force measurements and mathematical modeling to gain greater understanding of complex biological systems.
Imaging and force measurement at atomic resolution
0.1 nm special resolution under physiological conditions, dynamic changes, and temperature studies; 5 pN force and force modulation resolution.
- Multimode PicoForce AFM
- TIRF AFM (Bioscope Catalyst)
- Raman (LabRam) AFM (XE-120)
- Single Molecule Force Spectroscopy (SMFS) AFM (ForceRobot)
- Open-source platforms for nanotechnologies (AFM Workshop & developing High-speed AFM)
- Multi-modality (Raman, TIRF, NSOM, etc.)
- Environmental Control
- AFM Tip Functionalization
- Surface Modifications Staff
- Postdoc Fellow Visiting Fellow
- Staff Scientist
- Post-fusion structural changes and their roles in exocytosis and endocytosis of dense-core vesicles. Chiang HC, Shin W, Zhao WD, Hamid E, Sheng J, Baydyuk M, Wen PJ, Jin A, Momboisse F, Wu LG. Nat Commun. 2014.
- Acetylcholinesterase-catalyzed hydrolysis allows ultrasensitive detection of pathogens with the naked eye. Liu D, Wang Z, Jin A, Huang X, Sun X, Wang F, Yan Q, Ge S, Xia N, Niu G, Liu G, Hight Walker AR, Chen X. Angew Chem Int Ed Engl. 2013 Dec 23.
- VEGF-loaded graphene oxide as theranostics for multi-modality imaging-monitored targeting therapeutic angiogenesis of ischemic muscle. Sun Z, Huang P, Tong G, Lin J, Jin A, Rong P, Zhu L, Nie L, Niu G, Cao F, Chen X. Nanoscale. 2013 Aug 7.
- Gold nanoparticle-based activatable probe for sensing ultralow levels of prostate-specific antigen. Liu D, Huang X, Wang Z, Jin A, Sun X, Zhu L, Wang F, Ma Y, Niu G, Hight Walker AR, Chen X. ACS Nano. 2013 Jun 25.
- Development of a Pfs25-EPA malaria transmission blocking vaccine as a chemically conjugated nanoparticle. Shimp RL Jr, Rowe C, Reiter K, Chen B, Nguyen V, Aebig J, Rausch KM, Kumar K, Wu Y, Jin AJ, Jones DS, Narum DL. Vaccine. 2013 Jun 19.
- Further studies with isolated absolute infrared spectra of bacteriorhodopsin photocycle intermediates: conformational changes and possible role of a new proton-binding center. Hendler RW, Meuse CW, Smith PD, Kakareka JW. Appl Spectrosc. 2013 Jan.
- Unraveling protein-protein interactions in clathrin assemblies via atomic force spectroscopy. Jin AJ, Lafer EM, Peng JQ, Smith PD, Nossal R. Methods. 2013 Mar.
- Fabrication of hydrogels with steep stiffness gradients for studying cell mechanical response. Sunyer R, Jin AJ, Nossal R, Sackett DL. PLoS One. 2012.
- A nanoscale graphene oxide-peptide biosensor for real-time specific biomarker detection on the cell surface. Wang Z, Huang P, Bhirde A, Jin A, Ma Y, Niu G, Neamati N, Chen X. Chem Commun (Camb). 2012 Oct 9.
- Nuclear mapping of nanodrug delivery systems in dynamic cellular environments. Bhirde AA, Kapoor A, Liu G, Iglesias-Bartolome R, Jin A, Zhang G, Xing R, Lee S, Leapman RD, Gutkind JS, Chen X. ACS Nano. 2012 Jun 26.
- Analysis of the conformation and function of the Plasmodium falciparum merozoite proteins MTRAP and PTRAMP. Uchime O, Herrera R, Reiter K, Kotova S, Shimp RL Jr, Miura K, Jones D, Lebowitz J, Ambroggio X, Hurt DE, Jin AJ, Long C, Miller LH, Narum DL. Eukaryot Cell. 2012 May.
- A hematoma detector-a practical application of instrumental motion as signal in near infra-red imaging. Riley JD, Amyot F, Pohida T, Pursley R, Ardeshirpour Y, Kainerstorfer JM, Najafizadeh L, Chernomordik V, Smith P, Smirniotopoulos J, Wassermann EM, Gandjbakhche AH. Biomed Opt Express. 2012 Jan 1.
- Functional MnO nanoclusters for efficient siRNA delivery. Xing R, Liu G, Quan Q, Bhirde A, Zhang G, Jin A, Bryant LH, Zhang A, Liang A, Eden HS, Hou Y, Chen X. Chem Commun (Camb). 2011 Nov 28.
- Infrared and visible absolute and difference spectra of bacteriorhodopsin photocycle intermediates. Hendler RW, Meuse CW, Braiman MS, Smith PD, Kakareka JW. Appl Spectrosc. 2011 Sep.
- Enhanced mechanical rigidity of hydrogels formed from enantiomeric peptide assemblies. Nagy KJ, Giano MC, Jin A, Pochan DJ, Schneider JP. J Am Chem Soc. 2011 Sep 28.
- Quantitative principal component model for skin chromophore mapping using multi-spectral images and spatial priors. Kainerstorfer JM, Riley JD, Ehler M, Najafizadeh L, Amyot F, Hassan M, Pursley R, Demos SG, Chernomordik V, Pircher M, Smith PD, Hitzenberger CK, Gandjbakhche AH. Biomed Opt Express. 2011 Apr 1.
- AFM visualization of clathrin triskelia under fluid and in air. Kotova S, Prasad K, Smith PD, Lafer EM, Nossal R, Jin AJ. FEBS Lett. 2010 Jan 4.
- Structure of the Plasmodium falciparum circumsporozoite protein, a leading malaria vaccine candidate. Plassmeyer ML, Reiter K, Shimp RL Jr, Kotova S, Smith PD, Hurt DE, House B, Zou X, Zhang Y, Hickman M, Uchime O, Herrera R, Nguyen V, Glen J, Lebowitz J, Jin AJ, Miller LH, MacDonald NJ, Wu Y, Narum DL. J Biol Chem. 2009 Sep 25.
- Nanoscintillator conjugates as photodynamic therapy-based radiosensitizers: calculation of required physical parameters. Morgan NY, Kramer-Marek G, Smith PD, Camphausen K, Capala J. Radiat Res. 2009 Feb.
- Simultaneous measurements of fast optical and proton current kinetics in the bacteriorhodopsin photocycle using an enhanced spectrophotometer. Kakareka JW, Smith PD, Pohida TJ, Hendler RW. J Biochem Biophys Methods. 2008 Apr 24.