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Thursday, 28 February 2013 11:11

Membrane and Cytoplasm Calcium Neuronal Imaging

Neuronal Response to Caffeine Using a Chemical Probe in SPM 

 

 
Functional Calcium Imaging - An Optical Alternate to Patch Clamping

 

  • In this series of images a Nanopipette probe containing OrangeGreen is held in place with AFM control over a neuronal cell membrane of a cell which is filled with Fluo3.
  • The probe tip and the cell are excited in epi-illumination.
  • Each frame in the image is one confocal scan with 250 millisecond intervals between frames.  
  • A puff of caffeine is injected just before the fourth image (top row).
  • The cell and tip respond to the caffeine peaking in the center of the second row. Some of the pixels are red indicating that calcium has flowed out of the membrane and can be found at the tip which is in close proximity to the cell membrane.
  • Only Nanonics produces such probes for chemical imaging.
     
Thursday, 28 February 2013 11:05

Monkey's Kidney

Monkey Kidney Cells 
Fluorescence NSOM Images
  
 

     
35x35 micron AFM image    35x35 micron Transmission FluorescenceNSOM image

These images were produced using in the Intermittant Contact mode in a liquid environment using the Nanonics Liquid Cell with the MultiView 1000 system.

 

These topographic and fluorescence images were obtained simultaneously with cantilevered NSOM fiber optic probes. Labelled with rabbit anti-fibronectin primary antibody which was followed by a goat anti-rabbit Alexflour 488 secondary antibody. The Alexaflour probe is excited at about 488 and emits approximately 520nm. Thus, all fibronectin should be fluorescent. Fibronectin is part of the extracellular matrix which attaches the cell to the glass substrate.

The cell is visible in the AFM topography image but the fluorescent image shows mainly fibers. The fibers are primarily under the cell and therefore are difficult to image with AFM unless the cell is thin enough and these cells are thick since they were imaged at a more advanced stage in their growth. However, the fibers are fluorescent so they show up in the fluorescence NSOM optical image. 

Thursday, 28 February 2013 10:05

Current Mapping of Au Electrodes on Silicon

 

   
 AFM height Image of gold electrodes on Silicon substrate  Current mapping correlated image performed with MV4000 tuning fork feedback shows conductivity distribution of the sample shown below.

 

 

  sample description 
   

 

 

Performed with Nanonics unique ultrastable Nano-wire glass insulated electrical probe.

 

  • Ultrastable solid wire electrical probes q Ultra-Sensitive Tuning Fork  Normal Force feedback
  • Geometrical friendly for online multiprobe all under active AFM feedback.
  • Low contact resistance and full insulation with glass upto the probe tip for high electro-potential resolution
  • Glass coating insulation can be overcoated with metal to emulate coax geometries for ultrahigh sensitivity electrical imaging
  • High cantilever design that minimizes cantilever electrical interference

 

 
 
 Left: AFM height image of gold electrodes on silicon substrate performed with Multiview 4000TM using Tuning Fork intermittent feedback.

Right: Electrical image shows current conductivity on the gold electrodes. The non- continuity points are due to dust accumulated on the surface. 

  

Performed with Nanonics unique ultrastable Nano-wire glass insulated electrical probe

 

  • Ultrastable solid wire electrical probes q Ultra-Sensitive Tuning Fork  Normal Force feedback
  • Geometrical friendly for online multiprobe all under active AFM feedback.
  • Low contact resistance and full insulation with glass upto the probe tip for high electro-potential resolution
  • Glass coating insulation can be overcoated with metal to emulate coax geometries for ultrahigh sensitivity electrical imaging
  • High cantilever design that minimizes cantilever electrical interference
Thursday, 28 February 2013 10:01

Au etched groove on silicon

 
 
Left: AFM Height image of an etched groove of Au surface on Silicon substrate performed with two probe system.

Right: Electrical conductivity image performed with one a scanned probe while a second SPM probe is used to provide a nanometric local bias electrode.

 
Thursday, 28 February 2013 09:56

Protein Writing on Si Surface

 

BSA protein writing on Si surface using the Fountain Pen Nanolithography technique. On the left is an AFM image presenting two lines of the BSA. The plot on the right shows a height line profile of these lines. A pipette of 100nm aperture has been used with contact mode of 20µm /ms writing speed.

WSxM software has been used for image processing of the pictures above: I. Horcas et al. Rev. Sci. Instrum. 78, 013705 (2007). 
 

A silicon surface has been selected for writing due to its significance in the semiconductor industry. Using FPN for patterning on silicon could open the door for this technique to be integrated into different fields of semiconductors and materials.

Patterning with biological materials on silicon demonstrates the ability of FPN in biochip technology. FPN allows for patterning with biological materials onto Si surfaces, commonly used in integrated circuits and semiconductor devices.

Ideal systems for this application:

MV4000
MV2000
MV1000

 

Thursday, 28 February 2013 09:50

Protein Nanoprinting

 
     
 40 x 40 micron Topgraphy  

3D image

 

 

   
 
60 x 80 micron optical image of the same region.
 
 
Video of protein printing in real time
     

  
Only Nanonics can deliver chemicals or gas onto the sample on line, with no need to remove the tip from the sample.

Protein printing is made possible by the Nanonics Chemical Delivery System and Nanopipette

Using any of the Nanonics MultiView systems, chemicals, in liquid or gas form, can be fed into the pipette via a silicon tube. Apart from Protein Printing, applications of this setup include metallic nano-etching and nanolithography: an etchant can be introduced into the sample and scanned across it with nanometer precision using our 3D FlatScanning™ technology. The nanopipette is engaged by the sample using standard contact-mode atomic force microscopy, and our integrated system makes it possible to view the etching simultaneously through any optical microscope.


 

    Nanofountain Pen reviewed by Nature Materials

In the August 2003 edition of Nature Materials. The Nanofountain Pen was reviewed in the Material Update section.

Click to open application note on the uses of the nanofountain pen (331kB)

Thursday, 28 February 2013 09:48

AFM Imaging of TMP

 

X: 3.0 μm

 

X: 3.0 μm

 

Thursday, 28 February 2013 09:43

AFM Image of Tip

Standard Silicon Tip 
Imaged by an Optical Fiber AFM Probe

   
 
2D AFM image of the tip area approximately 15µm x 15µm
     
3D Topographic Image of standard Silicon tip measured by AFM in the intermittent contact mode of operation  
 
These images were produced by the Nanonics MultiView 1000™ with an optical fiber probe in the intermittent contact mode. 

Only the Nanonics optical fiber AFM probes have high enough aspect ratios (10:1) and are narrow enough to image the structure of a standard Silicon Probe.
 
 
Cantilevered AFM Probe: note that the angle of the cantilever enables viewing the sample underneath the AFM tip
 
 
Sketch of  AFM Probe
Thursday, 28 February 2013 09:38

Silicon NanoIndentation

 

 

Figure 1: NanoIndentation obtained on SiO2 thin layer in Si obtained with 50nm Berkovich Diamond tip.

A) AFM topographic image of a nanoindentation obtained with a load of 100µN.

B) Line profile crossing the nanoindentation areas shows a depth of 36.5nm.

 

C) AFM topographic image of a nanoindentation obtained with a load of 50 µN.

D) Line profile crossing the nanoindentation areas shows a depth of 15.08nm.

WSxM software has been used for image processing of the pictures above: I. Horcas et al. Rev. Sci. Instrum. 78, 013705 (2007)

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