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N-SIM E and STORM 4.0

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by

Pablo Hernandez-Varas

on 12 May 2016

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Transcript of N-SIM E and STORM 4.0

Breaking the diffraction limit
2x resolution improvement
over widefield


XY resolution 115 nm
Z resolution 270 nm
8x smaller volume
N-SIM
Nikon's super-resolution systems:
Nikon's
SIM
technology,
now
easier
Widefield
SIM
Detail enhancement
One-click-away
multicolor
imaging
How?

SIM principle
a)
Sample with high frequencies
that cannot be picked up by the microscope

b) Known illumination
pattern
(grid)

c) Interference patterns (
Moire fringes
) are generated when the illumination pattern is impossed on the sample
Super-resolution solutions

SIM technology
STORM

3D-SIM
sectioning power
renders striking Z resolution
N-STORM
Nikon's super-resolution systems:
Powerful
3D rendering
engine built in the latest
NIS-Elements

software version
SIM enables large
signal to noise
improvement
Ceded by Romain Guiet, EPFL, Switzerland, 2015
N-SIM live cell imaging capabilities are improved by
TuCam
Two camera-
simultaneous
imaging
Andor iXon3 897 EMCCD
Andor iXon Ultra EMCCD
Hamamatsu Orca Flash4.0 sCMOS
N-SIM E
(
E
asy)

Nikon's super-resolution systems:
By Incorporating a
488/561
dualband Dichroic, TuCam
doubles aquisition speed
Dedicated laser box
: 488, 561 and 640 nm
Multiband
extraflat
dichroics
exclusively manufactured by Nikon
Robust
3D-SIM
operation
Superior,
SR grade 100x
and
60x
optics
Sensitive sCMOS
Flash4.0
camera

New
SIM2.0
module
for image reconstruction


Thumbnail view
for
intuitive navigation
through reconstruction settings
Easy
evaluation
of image reconstruction quality by FFT
for more advanced users
FFT map view
FFT profile view
A new order
of magnitude in resolution
10x improvement

over widefield
XYresolution: 20 nm
Z resolution: 50 nm
How?

N-STORM principle
Single molecule localization
Dyes are pushed into the "
Dark State
"
Stochastic
dye molecule
blinking
leads to individual molecule localization
Thousands of imaging cycles lead to the
detection
of all molules
The coordinates for each blinking event can be located into a map, rendering a
super-resolution image
Unravel the information concealed in your samples
Widefield
N-STORM
Ceded by Romain Guiet, EPFL, Switzerland
Cylindrical lens for
3D
N-STORM
600 nm Z range
from a single N-STORM acquisition
Huang, Wang, Bates and Zhuang,
Science
, 2008
Reporter-based
multicolor
imaging
BSC-1 cells labelled for Tubulin (Alexa 647) and PMP70 (ATTO 488)

Mike Davidson, National High Magnetic Field Laboratory, The Florida State University
Nikon's super-resolution systems:
N-STORM
4.0

a new generation for N-STORM
New hardware,
fastest performance
New illuminator
Includes motorized
1x
,
2x
and
4x
magnification lenses for laser power concentration
New
100x HP lenses
,
required for handling high laser power while reducing chromatic aberrations
New
diaphragm
to protect the sample from bleaching
New
sCMOS
technology
camera
Orca
Flash4.0
from Hamamatsu
Up to 9 kW/cm2 for 647 nm laser light
Up to 80x80 microns FOV
High sensitivity
Take your
N-STORM experiments
from minutes to seconds!
Maximum
flexibility
to fit your application
Improved
warping
correction
Improved software for a more
powerful analysis
Previous
New
Accurate
drift correction
based in autocorrelation
Possibility to use fiducial markers
Before correction
After correction
New "
Fitting
overlapping peaks
" function...
...To cover gaps in areas with high dye density
STORM
4.0
Get more information out of your sample
Full transcript