Techniques
28 Jan 2010
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Techniques

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​​Here is an overview of the techniques available at the OCTOPUS facility. Please click on the title of the technique to find out further information and to read case studies. 

Technique and applications

Specifications

Stimulated emission depletion (STED) microscopy

Confocal microscopy: 
Multicolour imaging, Spectral imaging, Time-lapse imaging, 3D imaging, FRAP, FRET, FLIM, FCS, FCCS, FLCS, Mosaic stitching
STED nanoscopy:
3 colour STED imaging, 3D STED, STED FCS and FCCS, TCSPC
Analysis Software:
3D rendering and movie maker, deconvolution, object analysis, single particle tracking

Applications: 
-Biological research,
-Medical research,
-Drug discovery,
-Biophysics
-Physics,
-Chemistry
-Cell Biology (live cell imaging)
-Biosensors, and Biomarkers

Lasers: White light laser (80-10MHz) 470-680nm free choice, 592 and 775nm STED lasers


Detectors: 2x internal photon counting HyD, 2x external cooled HyD, 2x internal spectral PMTs


Objectives: 10x air, 20x IMM, 40x and 63x water, 100x oil immersion


Extra information: Scanner speed 1800Hz, Galvo Z for 4D, Rotation field of view, Adaptive Focus control.

(more...)

Structured illumination microscopy (SIM)

Technique: 
Super-resolution at half the diffraction limit in xyz axes.

Applications:
Biological and medical research, material science, chemistry

Lasers:Sequential multicolour imaging with 405, 488, 561, 642 nm excitation.

Detector: sCMOS

Objectives: 100x and 63x oil immersion, 63x water immersion

Extra information: z-stacks possible. 3 or 5 rotation angles

(more...)

Stochastical optical reconstruction and photo-activated localisation microscopy (STORM/PALM)

Technique: 
Single-molecule localisation super-resolution imaging. Down to 20 nm resolution in xy and 50 nm in z over 1.5 µm z range. z-resolution using phase ramp or biplane methods.
TIRF, HILO and epi-illumination.

Applications: 
Biological and medical research, material science, chemistry

Lasers: Multicolour imaging with 488, 561, 642, 750 nm excitation and 405 nm activation

Detectors: EMCCD and sCMOS cameras

Objectives: Vutara - 60x oil and silicone immersion, Elyra- 63x oil and water immersion,100x oil and silicone immersion

(more...)

Multicolour confocal laser scanning  

Technique: 
Multicolour excitation and emission. Both custom built and commercial, anisotropy

Applications: 
Biological research, Medical research, drug discovery, material science, geological samples

 

Lasers: 405 nm, 488 nm, 543 nm, 561 nm, 632 nm, Broad band NKT and Fianium , 470-900 nm combined with FLIM

Objectives: 4x, 10x, 40x air, 20x NA 0.75, 63x oil immersion, 60x water immersion NA 1.2, 60x air ELWD, 100x oil immersion NA 1.49

Extra information: Nikon EC1, EC2, Leica SP8, Custom with GSI Lumonics Galvos

(more...)

Multiphoton confocal-FLIM

Technique: 
Becker and Hickl SPC830, SPC150 and SPC230

Wide-field employing Andor ICCD, possible combination with TIRF

Applications:
Chemistry solution phase, biomaterial, advanced functional material, catalysis

 

Lasers: 405, 488, 543, 642nm laser wavelengths and white light

Objectives: 10x, 40x air, 20x NA 0.75, 63x oil immersion, 60x water immersion NA 1.2, 60x air ELWD, 100x oil immersion NA 1.49

Extra information: Fluorescence FLIM 25 ps-50 ns
Phosphorescence PLIM 100 ns-ms
Multi-time 25 ps – 100 ms combined FLIM-PLIM
Multi-colour 2-16 channels
Multi-wavelength excitation
Multi-spectral imaging.

(more...)

Confocal-Alternating Light Excitation (ALEX) Microscopy

Technique: 
Alternating Light EXcitation confocal microscope. Quantitative stoichiometry and FRET measurements of single molecules diffusing through a confocal volume.

Applications:
Biological research, drug discovery.

Predominantly used for quantifying conformational changes and ligand binding in single molecule protein structures.

Lasers: 532 nm Donor excitation
640 nm Acceptor excitation

Detectors: Perkin Elmer Single-Photon Avalanche Diodes

Objective: 100x oil immersion

Extra information: Measurable FRET range 3-8 nm.
Optics optimised for Cy3B-Atto647N FRET pair.

(more...)

Multidimensional single molecule microscopy  

Technique: 
Total Internal Reflection (SM-TIRF) Microscopy Widefield multidimensional fluorescence imaging where signal is limited to 100 nm layer adjacent to coverslip.

Applications:
Biological and medical research, material science, chemistry

Lasers: 405, 488, 532, 561, 642 nm or white-light excitation. Three-colour simultaneous imaging.

Detectors: Andor iXon EMCCDs.

Objectives: 100x TIRF oil immersion.

Extra information: Excitation and emission polarisation resolvable.

(more...)

Single molecule microscopy data analysis  

Technique: 
Advanced software pipelines and support for our range of single molecule microscopy techniques (more...)

Optical tweezers  

Technique: 
Laser capture of microscopic particles to measure forces, Raman spectroscopy and Mie scattering. Combined with imaging techniques of TIRF, confocal, multiphoton FLIM and PLIM

Applications:
Colloid and interface science, medical research, atmospheric chemistry, aerosol science, material science, biological research, plant cell studies, catalysis.

Lasers: Laser trapping: 488 nm, 514.5 nm, 532 nm, 700 – 900 nm, 1064 nm, 1090 nm.
Raman spectroscopy : 488 nm, 514.5 nm, 532 nm, 785 nm and 1064 nm.
Broadband Mie scattering: 400 nm to 1700 nm.

Detectors: 450 – 700 nm and 600 - 1000 nm Acton spectrographs\PI Spec 10 detectors
900 – 1700 nm Isoplane spectrograph\PI NIRvana detector.

Objectives: 100x and 63x oil immersion, 63x water immersion. 100x and 50x long working distance air.

(more...)

 


Contact: Martin-Fernandez, Marisa (STFC,RAL,CLF)