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TIRF/PALM/dSTORM Microscope

LS
05/05/2022
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Single-Molecule Super-Resolution Imaging

STORM reconstructs a super-resolution fluorescent image by combining precise localization information for individual fluorophores in complex fluorescent microscope specimens.
This powerful technology enables the visualization of molecular interactions at the nanoscopic level, opening up new worlds of scientific understanding.
TIRF TiE_1

Principle

Total Internal Reflection Fluorescence (TIRF) Microscopy

Various mechanisms are often employed in fluorescence microscopy applications to restrict the excitation and detection of fluorophores to a thin region of the specimen. Elimination of background fluorescence from outside the focal plane can dramatically improve the signal-to-noise ratio, and consequently, the spatial resolution of the features or events of interest. Total internal reflection fluorescence microscopy (TIRFM) exploits the unique properties of an induced evanescent wave or field in a limited specimen region immediately adjacent to the interface between two media having different refractive indices. In practice, the most commonly utilized interface in the application of TIRFM is the contact area between a specimen and a glass coverslip or tissue culture container.

The concepts underlying TIRFM are not new, and much of the recent interest in, and enthusiasm for, the technique have come about due to technological advances that facilitate its use. The availability of complete ready-to-use instrumentation systems for employment of the method, as well as developments in fluorophore technology, such as genetically encoded fluorescent species, have made it possible to investigate a number of cell membrane and other surface processes in a direct manner that was not previously possible.

Configuration

Objectives

Magnification   

Type   

Immersion

N.A

WD

 

100x

PlanApoTIRF

Oil

1.49

0.16-0.09

DIC

Filters

Name

BS

EM

GFP

ZT488rdc

ET525/50m

mCh

ZT561rdc

ET630/75m

GFP/mCh

ZT488/561rpc

ZET488/561m

DAPI/GFP/mCh/Cy5

ZT405/488/561/640rpc

ZET405/488/561/640m

Associated devices

Illumination Detector Software Z motor T°C & CO2 
ROPER laser bench
  • 405nm: 50mW
  • 491nm:100mW
  • 561nm:1000mW
  • 633nm: 300mW
Evolve512 EMCCD camera, Photometrics Metamorph TiE motor + PFS Cube & Brick, Life Imaging Service

Techniques

Applications

  • Live Cell Imaging
TIRF MICROSCOPE
n-storm-nikon
TIRF/PALM/STORM microscope