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The William Harvey Research Institute

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Imaging Facilities

Microvascular Research Live (cell/tissue) Imaging Suite (MIC-LIS) 

Our Centre houses:

  • LSM800 Laser Scanning Confocal Microscope with Aryscan
  • Single-Point Laser Scanning Confocal Microscopes (x3) allowing in vivo and in vitro time-lapse imaging as well as fixed sample imaging
  • Time-lapse epi-fluorescent Microscope (x1) for in vitro live cell imaging
  • Brightfield/epifluorescent axioscope microscopes (x2) for intravital microscopy
  • Image analysis Platforms (x2) with IMARIS™ or ZEN Blue ™ software.

LSM 880 Laser Scanning Confocal Microscope with AiryScan

This new state-of-the art, fast scanning (27 frames/sec @ 480 x 480 pixels) near super-resolution confocal microscope (max 140nm lateral and 400nm axial resolution) was installed in September 2016. 

It comprises an inverted, fully motorised (X,Y,Z) Zeiss Axio Imager 2 with 10x, 20x, 40x and 63x objectives, which supports epifluorescence (RGB) and brightfield microscopy. The stand is equipped with an incubation system with temperature and CO2 control making it possible to image living material in addition to fixed, sectioned biological material. 

The system is equipped with a Diode 405nm laser, a Multiline Argon laser (wavelengths 458nm, 488nm and 514nm), a DPSS 561nm laser, a HeNe 594nm laser and a HeNe 633nm laser. The confocal has a range of ultra-sensitive detectors, including the Airyscan detector module, that are run through the user-friendly ZEN software platform. The software includes modules for 3D/4D reconstruction and animation; spectral scanning; analysis and quantification of co-localisation; spectral unmixing, advanced tiling and photo-bleaching (FRAP, FRET).


This core confocal microscope facility was made possible by a grant from Barts Charity.


Single Point Laser Scanning Confocal Microscopes 

Zeiss 800 Laser Scanning Confocal Microscope (Inverted)

This inverted confocal microscope was installed in December 2015.

Light source: 4 solid-state laser diodes (405, 488, 561 and 640nm) and a transmitted light detector suitable for overlaying confocal fluorescence images with DIC images.

Detectors: Fitted with 2 ultra-sensitive GaAsP detectors with spectral detection functionality.

Available objectives:

  • EC Plan-Neofluarx, 10 (NA:0.3; WD=5.2), Dry
  • Plan-Apochromat, x20 (NA:0.8; WD=0.55 M27), Dry
  • Plan-Apochromat, x40 (NA:1.3; DIC UVVIS-IR), Oil
  • Plan-Apochromat, x63 (NA:1.4; DIC f/ELYRA), Oil 

Software: ZEN Blue ™ software interface

Other features include:

  • environmental control for temperature and CO2
  • definite focus hardware based drift correction
  • motorised stage suitable for both mark and find and tile scanning
  • Piezo z-drive for accurate high-speed z-sectioning
  • Charges: This system is currently charged at £25/hour or £100/night

Leica TCS SP5 & TCS SP8 up-right Laser Scanning Confocal Microscopes (up-right)

These 2 systems have upright configurations designed for intravital microscopy (VM) but are also able to cater for fast and highly sensitive multi-colour confocal imaging of fixed preparations such as whole-mounted tissues or cell culture samples. Both systems are laser point-scanning devices and will support 4D imaging applications. Both microscopes are configured to scan in two different modes:

1. Standard mode for higher resolution with speed of scanning between 400Hz to 1800Hz.

2. Rapid – Resonancemode for fast acquisition with a speed of scanning of 8000Hz (e.g. ~ 50 frames for 30 sec at a resolution of 1024 x1024 with 3 colours).

Leica systems use a prism/slit technology (e.g. adjustable band-pass filters) to finely tune the recovered wavelengths to your needs.

Available objectives:

  • x10, (NA:0.3), Dry
  • x20, (NA:1), Water dipping
  • x63, (NA:0.9), Water dipping
  • x63, (NA:1.4), Oil

Specific configurations of the two microscopes:

TCS SP5 

Light source: 7 laser lines (458, 476, 488, 496, 514, 561, 633 nm).

Detectors: Channel Spectral Scan-head with 3 SP detectors.

Software: LAS-AF version 3.3.

How to define the spectral bands that collect probe specific emission.  

TCS SP8

Light source: Freely tuneable (1 nm precision) White Light Laser able to generate up to 8 simultaneous excitation lines of 470 – 670 nm continuum.

Detectors: Channel Spectral Scan-head detectors, including two SP conventional detector and 2 Hybrid Detector HyD for high sensitivity/ low noise imaging.

Software: LAS-AF version 2.6.

Charges 
The use of these platforms is currently charged at £25/hour.

Further information 

Time-Lapse EPI-Fluorescent Microscope

This inverted Olympus IX81 microscope is fitted for bright-field and fluorescence imaging of live cells.

Available objectives:

  • x 4 Dry
  • x 10 Dry
  • x 20 Dry
  • x 40 Dry

Fluorescence detection filter set: DAPI/ FITC /TRITC.

Software: Cell M™.

Other features include: The system features environment-controlled chamber accommodating culture dishes and flasks and a fast and high-resolution camera allowing multi-position time-lapse imaging.

Charges: The time lapse epi-fluorescent microscope is currently charged at £100/per night.

Brightfield and EPI-Fluorescent Microscopes

These two upright Zeiss Axioscop microscopes are established for intravital microscopy of transparent tissues (e.g. cremaster muscles, mesentery) as well as epi-fluorescence imaging.

Light source: Halogen and HBO lamps.

Objectives:

Water dipping:

  • x 100 (NA:1)
  • x 20 (NA:0.5)
  • x 40 (NA:0.8) 

Dry:

  • x 10 (NA:0.25)
  • x 20 (NA:0.45)
  • x 40 (NA:0.65)

Filters sets: Green-emitting (GFP, A488, FITC) and red-emitting (TRITC, Cy3, A555) dyes.

Detectors: Hamamatsu Colour Chilled 3CCD, JVC colour 3CCD KY-F55B. 

One of the Axioscope microscopes can do DIC imaging and both are connected to a computer for visualisation and recording of live imaging videos.

Charges: The use of these platforms is currently charged at £10/hr.

4D/3D Image Analysing Platforms

  • Measurement Pro (for all type of measurements such as volumes area, spots detections, intensities)
  • Imaris Track (to track object parameters in 4D)
  • Imaris Coloc (for colocalisation studies)

We have 2 image analysis platforms.

(x1) with IMARIS™ software (version 7.3) including the following modules: 
(x1) with ZEN Blue ™ including the following modules:

  • Colocalisation, Deconvolution, 3D VisArt
  • Image Analysis & Interactive Measurement 

Charges: The Image analysis platforms can be accessed for a fee of £5/hr.

Further information.

With respect to all platforms, where significant advice and training is required, this will be provided on a collaborative basis with members of the Centre for Microvascular Research.

All this equipment is available for researchers within QMUL and beyond. We can provide support, advice and training for anyone interested in using our facility.

For enquiries regarding the use of these platforms, please contact:

Dr Mathieu-Benoit Voisin  - Email
Dr Tom Nightingale - Email

Location
John Vane Science Centre
4th floor, Microvascular Research
Charterhouse Square
London EC1M 6BQ

 


 

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