Atomic Force Microscope BSL-3

AFM / Optical Microscope (BIO-AFM) for L3 imaging of infected living cells and class 3 pathogens (viruses and bacteria). 

Possible samples:

fixed or living cells in their culture medium, bacteria, yeasts, macromolecules (nucleic acids, proteins), polymers, fibers, composites, lipid films, lipid vesicles (GUV, LUV), extracellular vesicles (exosomes), structures purified biologics (viral capsids for example), virus-like-particles (VLPs), class 2 and 3 viruses.

Size of the smallest objects detectable on the surface: a few nanometers X and Y

Height of observable objects: from 0.5nm to 12μm

Dimension of square images: from 50nm to 100μm

Sizes of the observable samples: 30mm x 50mm in X and Y, 5 to 10mm in Z

Possibility of working in liquid or air in a thermostatic chamber.



AFM images :

a) Chikungunya virus

b) Bacteria

c) HEK293T cell


Transportin-1 binds to the HIV-1 capsid via a nuclear localization signal and triggers uncoating.
Fernandez J, Machado AK, Lyonnais S, Chamontin C, Gärtner K, Léger T, Henriquet C, Garcia C, Portilho DM, Pugnière M, Chaloin L, Muriaux D, Yamauchi Y, Blaise M, Nisole S, Arhel NJ.
Nat Microbiol. 2019 Oct 14. doi: 10.1038/s41564-019-0575-6.

Figure: In vitro reconstituted HIV-1 proteins using recombinant capsid were contacted with purified Transportin. The images taken by Atomic Force Microscopy illustrate the deformations and breaks by HIV-1 capsid representing (left to right) 10, 20 and 40 min after injection.


Lethality of Brucellaï¿¿microti in a murine model of infectiondepends on the wbkE gene involved in O-polysaccharide synthesis

Safia Ouahrani-Bettache, María P. Jiménez De Bagüés, Jorge De La Garza,Luca Freddi, Juan P. Bueso, Sébastien Lyonnais, Sascha Al Dahouk, Daniela De Biase, StephanKöhler & Alessandra Occhialini. 2019 , Virulence, 10:1, 868-878, DOI:10.1080/21505594.2019.1682762

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AFM Nanowizard 4 head (JPK-Bruker) customised for L3 environment (no sharp edges, sealing against liquids and vapors)

980 nm laser source

imultaneous fluorescence and AFM imaging

Fast scan mode (up to 70Hz / line for scans up to 2μm)

Imaging methods in air or in liquid:

  • Quantitative QI â„¢ Imagingo Side force contact mode
  • AC mode with phase contrast, force or frequency modulation
  • Force spectroscopy & elasticity methods: unique force curves, force-mapping

XY scan: 100μm x 100μm,

Z scan: 15μm-

BSL3 (JPK PetriDishHeater) heated sample holder (up to 60 ° C) for 35x10mm petri dishes (WPI and TPP, plastic or plastic with glass bottom) or 25 mm glass strips.- S

Optical Microscope NIKON Ti2-U inverted Microscope with Phase Contrast and Fluorescence

  • Nikon CFI Lens Achromat LWD ADL-20x Ph1-
  • Nikon CFI Achromat LWD ADL-40x Ph2
  • NIKON CFI APO VC 100x oil (A.N. 1.4 DT 0.13)

White LED light source and Epi-Fluorescence Intensilight 100w (Hg, manual)

  • Cubes Filters:o GFP (Ex466 / 40, DM 495, stop filter BA 525/50)
  • mCHERRY, mRFP (Ex562 / 40, DM 593, BA 640/75 stop filter)
  • CY5, APC, DiD, Alexa Fluor 647, Alexa fluor 660 (Ex628 / 40, DM 660, BA 692/40 stop filter)

JENOPTIK Fluorescence ProgRes MFcool Camera (2/3 “1.4 MegaPixel CCD, 1350×1024 pixel resolution, 14bit)