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

​​Multimodality Optical NanoScopy TERminus (MONSTER)​​

​​​​Technique:
Confocal, Airyscan, Structured Illumination Microscopy, Single Molecule Localisation Microscopy (PALM/STORM).

Applications:
Biological and medical research, material science, chemistry

Lasers: Multicolour imaging with 405, 458, 488, 514, 561, 6330 nm excitation.

Detectors: Dual EMCCD, AiryScan detector.

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

Lightsheet Microscopy​
Technique:
Light sheet fluorescence microscopy on Leica SP8

Additional techniques:

Confocal microscopy
Multiphoton (MP) and non-linear microscopy
Fluorescence lifetime imaging (FLIM)
MP FLIM
FCS

Applications:
Biological and medical research Developmental biology Real time live imaging Large samples – spheroids, tissues, small embryos
Lasers:
White light laser (80-10MHz) 470-680 nm free choice
405 nm CW
Ti:Sapph for MP 80 MHz 680-1300 nm free choice

Detectors:
Hamamatsu FLASH 4 Camera,
2 internal photon counting HyD, 2 internal PMTs, 2 external cooled HyD, 2 external PMTs, trans-PMT

Objectives: 
Light sheet detection: 5x W, 10x W, 25x W 
Confocal and multiphoton: 63X W, 63X Oil, 40X W, 25X IR W, 10X, 5X

Extra information:
Leica TCS SP8 DLS​ 

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...)

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)