Atomic Force Microscopes : Bio-AFM
Atomic Force Microscopes (AFMs) are a type of Scanning Probe Microscope (SPM) that map surfaces by measuring interactions between a tiny probe and the sample. BioAFMs extend this capability by integrating fluorescence optics with an inverted microscope, enabling high-resolution imaging of biological samples in liquid. This makes them ideal for studying tissues, cells, hydrogels, DNA, and proteins in near-physiological conditions.
Getting Started and Training Information
Not sure if the bioAFM is the right tool for your project? Reach out to Email SPM Staff to schedule a quick chat.
If you would like to request a staff run BioAFM measurement on your sample, please fill out this intake: Stanford University BioAFM Qualtrics Form
Staff-run measurements can include high-resolution imaging of sample surface features or mechanical property mapping, depending on your project needs. Basic data post-processing will also be provided.
To become a qualified user of the bioAFM, follow these steps:
1) iLab Account: Ensure you have an active iLab account to reserve the instrument. If you don't have one, set up your account here. CSIF iLab Account Registration
2) Complete the edX AFM Training course:
- Join edX and take all of the parts of the Atomic Force Microscopy online training course, including all 4 quizzes.
i) Click on this LINK and search for the nano@stanford course.
ii) Navigate to the Atomic Force Microscopy section and take the required modules, read the content, and take the quizzes.
- New User Intake Form: Complete the SPM New User Intake Form: Click this link: SPM New User You will be contacted shortly to set up your training.
Training typically consists of four 3-hour sessions, with the opportunity to bring your own sample in for the final session:
Session 1: Overview of AFM fundamentals and a demo showing setup and imaging in air.
Session 2: Demo showing setup and imaging in liquid.
Session 3: Hands-on review of session 2.
Session 4: Trainee brings in sample and sets up imaging under supervision.
CSIF houses two Atomic Force Microscopes (AFMs):
JPK NanoWizard 5 BioAFM
1. NanoWizard V NWV BioAFM
The JPK NanoWizard V (NWV) AFM available at CSIF is a state-of-the-art instrument that combines high spatio-temporal resolution with a large scan area, intuitive software, automated setup steps, a comprehensive suite of the latest imaging modes, and seamless integration with optical microscopy. The NWV excels at performing in situ measurements in liquid environments with correlative optical imaging. The versatile NWV is equipped to measure high resolution topography and mechanical properties of samples ranging from thin films and polymers to tissue sections, cells and biomolecules such as DNA. The NWV can also image dynamic molecular events, from molecular dynamics of proteins to kinetics of crystal growth and dissolution!
Features:
● Automated setup steps for fast and easy operation with fully motorized laser and detector alignment.
● Large sample scan range of up to 200x200μm² with HybridStage.
● Easily obtain mechanics of sticky, corrugated, or rough samples such as polymers, tissues, and hydrogels with a Z range of up to 200μm.
● Easy correlative optical and AFM measurements for both transparent/non-transparent samples.
● AFM integration with either: Inverted Microscope, Zeiss AxioObserver: 10x, 20x, 40x phase, 63x oil, and 100x oil objectives with GFP, RFP, and DAPI fluorescent channels
● Stereo Microscope, Zeiss AxioZoom: 0.5X with GFP, RFP, and DAPI fluorescent channels.
● Excels at in situ liquid measurements with sample heating (up to 60°C), perfusion, and CO2 control (from 0% to 20% volume) capabilities. High resolution imaging in air.
● Image dynamics molecular events with the fast scan capability: scan speed up to 200 lines/sec with up to 1.5μm height range.
● Complete suite of AFM modes for topography, mechanics, surface properties, force spectroscopy, and viscoelastic measurements:
○ PeakForce-QI
○ PeakForce Tapping®
○ QI mode
○ TappingMode™ with PhaseImaging™
○ Contact Mode with Lateral Force
○ Fast Force Mapping
○ Force spectroscopy
● Dedicated viscoelastic property mapping software for sine modulation measurements at defined frequencies.
● Equipped with Förster resonance energy transfer (FRET) capabilities to use FRET as a sensor for real-time in-situ proximity monitoring. With GFP/RFP and CFP/YFP pairs.
If you use this instrument in your research, please include the following acknowledgment in your publications:
“The authors acknowledge the Stanford University Cell Sciences Imaging Core Facility (RRID:SCR_017787) for access to instrumentation. This work was supported by the Air Force Office of Scientific Research (AFOSR) through DURIP Grant FA9550-24-1-0063.”
Contact and Location
Nadia Makarova nmak@stanford.edu, Shriram Center, Room B023
2. Bruker BioScope Resolve BioAFM
- Resolving Nanoscale Structures on Living Cells
- Molecular High-Resolution Imaging
- Real-Time Cell Topography, Modulus, and Fluorescence Observation using Zeiss LSM900 AiryScan
- Quantitative Cell Mechanics and Molecular Force Spectroscopy Data
If you use this instrument in your research, please include the following acknowledgment in your publications:
“The authors acknowledge the Stanford University Cell Sciences Imaging Core Facility (RRID:SCR_017787) for access to instrumentation. This work was supported by the National Institutes of Health (NIH) Office of Research Infrastructure Programs (ORIP) under Grant S10 OD021514-01.”
Contact and Location
Nadia Makarova nmak@stanford.edu, Shriram Center, Room B023