Biomod/2011/TeamJapan/Sendai/Result Atomic Force Microscope/detail Observation 2D and 3D structure

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Older design of the 2D and 3D robot

We used High-speed AFM(RIBM, Nano Live Vision).
silicon nitride cantilever:
  • Olympus BL-AC10EGS-A2
resonant frequency = 1.0-2.0 MHz
spring constant = 0.1-0.3 N/m
EBDTip radius < 15 nm
  • Olympus BL-AC10DS-A2
resonant frequency = 1.5 MHz
spring constant = 0.1 N/m
EBDTip radius = 24nm
Scale bar : Figure.1-26, 200 nm

 Old2D 

2D_noncut
2D_noncut


This is a figure of 2D structure before cutting M13. There is surplus M13(about 5000 bp) from the middle of the 2D structure since whole of M13 was used.


  • Condition

Buffer:1×TAE
Anealing time:2 hours
m13:staple=1:100(ratio of concentration)


When Nickel was Put in,We observed AFM pictures clearly.
However, Nickel makes agregation increase while it makes adsorption power of mica heighten.
So we dicided not to use Nickel.


We confirmed M13:staple=1:10(ratio of concentration)
In this concentration,it is easy to observe structures since agregation decrease.

 Old3D 

We can't observe the 3D structure using whole of M13.
As this factor,there are 3 points.

(1) The strucuture was not able to fold because of surplus M13.
(2) It was difficult to confirm the 3D structure in hispeed AFM.
(3) The 3D structure is similar to small agregation.

To observe the 3D structure,We need

(1)Few surplus M13(We succeeded after this experiment)
(2)Purification skill of agregation


  • Condition

Buffer:1×TAE
Anealing time:2 hours

m13:staple=1:10(ratio of concentration)

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