BIXAM is a new imaging system under development that combines inverted optical microscopy/confocal scanning laser microscopy and atomic force microscopy (AFM).
It allows for simultaneous observation using optical microscopy fluorescence imaging and AFM video imaging.

The BIXAM’s high resolution images.

New discoveries are being published with the images captured by the BIXAM

New discoveries are being published with the images captured by the BIXAM
An improved unroofing method enabled the BIXAM to view intracellular cytoskeletal actin filaments
in phosphate- buffered saline at a high resolution.
Images courtesy of the Structural Biology Research Center, Nagoya University, Japan
(Professor Usukura and Associate Professor Narita).(*5-1).
“An Unroofing Method to Observe the Cytoskeleton Directly at Molecular Resolution
Using Atomic Force Microscopy“, Eiji Usukura et al. Scientific Reports, 27472 (2016)
doi:10.1038/srep27472 opens in new window
(A)A BIXAM image of actin as a control.
(B)One example of the actin –tropomyosin images.
(A)A BIXAM image of actin as a control. There was an average of nine frames.
(B)One example of the actin –tropomyosin images. There was an average of seven frames.
Images courtesy of the Structural Biology Research Center, Nagoya University, Japan
(Associate Professor Narita and Professor Usukura).(*5-2).
"Direct Observation of the Actin Filament by Tip-scan Atomic Force Microscopy,"
Akihiro Narita et al., Microscopy (Tokyo)(2016)
doi: 10.1093/jmicro/dfw017 opens in new window
(A)Schematic representation of mature DE-cadherin
(B) Representative images of DEEC6-His(above) and DEEC6-GFP-His (below).
White arrows in B point to the presumed GFP portions.
(C) High resolution image (above) and interpretive illustration (below)
(D) Course over time and schematic illustration of a DEEC6-His molecule whose head portion turned.
Yellow arrows in D point to the N-terminal. Scale : 10nm.
Images courtesy of the Department of Biological Sciences, Graduate School of Science,
Osaka University, Japan (Associate Professor Oda)
"Divergence of Structural Strategies for Homophilic E-cadherin Binding Among Bilaterians,"
Shigetaka Nishiguchi et al., Journal of Cell Science (2016)
doi: 10.1242/jcs.189258 opens in new window

Allows for simultaneous video observation using an optical microscope and AFM !

Example of Simultaneous Observation using Confocal Microscope and AFM
Clathrin and caveolin in a living COS7 cell are fluorescent-labeled.
Observing multiple luminescent spots and endocytosis pits simultaneously.
Overlay of an AFM image on a confocal microscope image of
mitochondria in a living COS7 cell (still picture)
Overlay of an AFM image on a confocal microscope image of mitochondria in a living COS7 cell (still picture)
  • AFM imaging of the SURFACE of living cells Observing dynamics on the cell membrane (endocytosis)
  • AFM imaging of the INSIDE of living cells Observing organelle (actin)dynamics through the cell membrane(*3)

TIRF and high-resolution fluorescence microscopes, such as super-resolution microscopes, are often used in the fields of life science, serving as powerful tools for investigating the fluorescent-labeled sites of living cells. They have a resolution that can zoom down to around 20 nm, detect individual molecules, and provide location information.

On the other hand, the conventional AFM is powerful equipment that allows for the highspeed observation of sample morphologies at a resolution of 1 nm or less. However, it is incapable of identifying substances observed. While the AFM is suitable for observing molecules, such as purified proteins with a known structure, it has been difficult to identify the substances observed in samples where various substances co-exist (such as the membrane surfaces of living cells).

BIXAM is a new item of observation equipment that combines the advantages of inverted optical microscopy/confocal scanning laser microscopy and AFM. It allows for the detailed observation of morphologies and events in specific sites by fluorescent labeling and by further observation of the sites using AFM.

Observing Cultured Cells

Powerful system for observing the dynamic behavior of non-fixed cultured cells

Observing endocytosis (A)and undulation (D)etc., on a cell membrane
Observing actins (B)and mitochondria (C)etc., through a cell membrane (without any membrane breakage)

Observing Cultured Cells

  • (A)Endocytosis in a living NRK cell
  • (B)Actin filaments in a living NRK cell
  • (C)Simultaneous observation of a living COS7 cell using confocal fluorescence microscopy and AFM
  • (D)Endocytosis and pseudopodia undulation in a living COS7 cell(*4)

Observing the dynamics of biomolecules on a lipid bilayer membrane

Excellent for observing the dynamics of biomolecules on a mica-supported lipid bilayer(*4)

  • Observing DNA origami

Also ideal for still images

Also powerful for still images
To prepare for observation simply place your sample on a microscope slide

Observing unroofed cells(*5-1)

  • A series images of a clathrin coat in (A) to (C)
captured in every 20 seconds.
    A series images of a clathrin coat in (A) to (C) captured in every 20 seconds. *5-1
  • Caveorae
    Caveorae *5-1

Observing Purified Biomolecules

  • DNA double helix
    DNA double helix
  • Two-dimensional streptavidin crystals
    Two-dimensional streptavidin crystals

GalleryBIXAM video image gallery

  • BIXAM
  • BIXAM

Current Performance of BIXAM prototype

BIXAM features Simultaneous video observation of an optical microscope and AFM
Optical microscopy observation An inverted optical microscope/a confocal scanning laser microscope
AFM characteristics Video observation of living cells etc., using cantileverscanning in an aqueous environment
Sample substrate Slide
Observed rate of AFM 0.05~10fps(frames/second)
Observation range of AFM X:3µm or more, Y:2µm or more, Z:1µm
Cantilever USC-f0.8-k0.1-T12(NanoWorld AG)
BL-AC10FS; BL-AC10DS
Observation environment Aqueous
Observation modeof AFM AC mode

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