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.

Available equipment

  • 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)

Technologies

  • Multi-modality (Raman, TIRF, NSOM, etc.)
  • Environmental Control
  • AFM Tip Functionalization
  • Surface Modifications Staff
Yu GYang ZFu XYung BCYang JMao ZShao LHua BLiu YZhang FFan QWang SJacobson OJin AGao CTang XHuang FChen X
Nat Commun
2018 Feb 22

Zhu GLynn GMJacobson OChen KLiu YZhang HMa YZhang FTian RNi QCheng SWang ZLu NYung BCWang ZLang LFu XJin AWeiss IDVishwasrao HNiu GShroff HKlinman DMSeder RAChen X
Nat Commun
2017 Dec 05

Zhu GMei LVishwasrao HDJacobson OWang ZLiu YYung BCFu XJin ANiu GWang QZhang FShroff HChen X
Nat Commun
2017 Nov 14

Bhat AEdwards LWFu XBadman DLHuo SJin AJLu Q
Appl Phys Lett
2016 Dec 26


Wen PJGrenklo SArpino GTan XLiao HSHeureaux JPeng SYChiang HCHamid EZhao WDShin WNäreoja TEvergren EJin YKarlsson REbert SNJin ALiu APShupliakov OWu LG
Nat Commun
2016 Aug 31

Sousa RLiao HSCuéllar JJin SValpuesta JMJin AJLafer EM
Nat. Struct. Mol. Biol.
2016 Sep


Zhu GLiu YYang XKim YHZhang HJia RLiao HSJin ALin JAronova MLeapman RNie ZNiu GChen X
Nanoscale
2016 Mar 28


Rong PHuang PLiu ZLin JJin AMa YNiu GYu LZeng WWang WChen X
Nanoscale
2015 Oct 21

Huang PGao YLin JHu HLiao HSYan XTang YJin ASong JNiu GZhang GHorkay FChen X
ACS Nano
2015 Oct 27

Herrera RAnderson CKumar KMolina-Cruz ANguyen VBurkhardt MReiter KShimp RHoward RFSrinivasan PNold MJRagheb DShi LDeCotiis MAebig JLambert LRausch KMMuratova OJin AReed SGSinnis PBarillas-Mury CDuffy PEMacDonald NJNarum DL
Infect. Immun.
2015 Oct


Chiang HCShin WZhao WDHamid ESheng JBaydyuk MWen PJJin AMomboisse FWu LG
Nat Commun
2015 Jan 27

Yan XHu HLin JJin AJNiu GZhang SHuang PShen BChen X
Nanoscale
2015 Feb 14

Yan XNiu GLin JJin AJHu HTang YZhang YWu ALu JZhang SHuang PShen BChen X
Biomaterials
2015 Feb


Huang PRong PJin AYan XZhang MGLin JHu HWang ZYue XLi WNiu GZeng WWang WZhou KChen X
Adv. Mater. Weinheim
2014 Oct 08

Choi KYSilvestre OFHuang XMin KHHoward GPHida NJin AJCarvajal NLee SWHong JIChen X
ACS Nano
2014 May 27

Bhirde AAChikkaveeraiah BVSrivatsan ANiu GJin AJKapoor AWang ZPatel SPatel VGorbach AMLeapman RDGutkind JSHight Walker ARChen X
ACS Nano
2014 May 27

Chiang HCShin WZhao WDHamid ESheng JBaydyuk MWen PJJin AMomboisse FWu LG
Nat Commun
2014

Liu DWang ZJin AHuang XSun XWang FYan QGe SXia NNiu GLiu GHight Walker ARChen X
Angew. Chem. Int. Ed. Engl.
2013 Dec 23


Liu DHuang XWang ZJin ASun XZhu LWang FMa YNiu GHight Walker ARChen X
ACS Nano
2013 Jun 25

Shimp RLRowe CReiter KChen BNguyen VAebig JRausch KMKumar KWu YJin AJJones DSNarum DL
Vaccine
2013 Jun 19



Wang ZHuang PBhirde AJin AMa YNiu GNeamati NChen X
Chem. Commun. (Camb.)
2012 Oct 09

Bhirde AAKapoor ALiu GIglesias-Bartolome RJin AZhang GXing RLee SLeapman RDGutkind JSChen X
ACS Nano
2012 Jun 26

Uchime OHerrera RReiter KKotova SShimp RLMiura KJones DLebowitz JAmbroggio XHurt DEJin AJLong CMiller LHNarum DL
Eukaryotic Cell
2012 May

Xing RLiu GQuan QBhirde AZhang GJin ABryant LHZhang ALiang AEden HSHou YChen X
Chem. Commun. (Camb.)
2011 Nov 28

Nagy KJGiano MCJin APochan DJSchneider JP
J. Am. Chem. Soc.
2011 Sep 28

Bhirde AALiu GJin AIglesias-Bartolome RSousa AALeapman RDGutkind JSLee SChen X
Theranostics
2011

Kotova SPrasad KSmith PDLafer EMNossal RJin AJ
FEBS Lett.
2010 Jan 04

Plassmeyer MLReiter KShimp RLKotova SSmith PDHurt DEHouse BZou XZhang YHickman MUchime OHerrera RNguyen VGlen JLebowitz JJin AJMiller LHMacDonald NJWu YNarum DL
J. Biol. Chem.
2009 Sep 25

Tsai CWDuggan PFJin AJMacdonald NJKotova SLebowitz JHurt DEShimp RLLambert LMiller LHLong CASaul ANarum DL
Mol. Biochem. Parasitol.
2009 Mar

Hayakawa ETokumasu FNardone GAJin AJHackley VADvorak JA
Biophys. J.
2007 Dec 01

Jin AJPrasad KSmith PDLafer EMNossal R
Biophys. J.
2006 May 01

Forbes JGJin AJMa KGutierrez-Cruz GTsai WLWang K
J. Muscle Res. Cell. Motil.
2005



Tokumasu FJin AJFeigenson GWDvorak JA
Biophys. J.
2003 Apr