Imaging Facility

Imaging facilityThe Imaging section is now equipped with 4 confocal microscopes including two recently acquired additions, a Zeiss 780 Zen which is equipped for live cell imaging. Also we now have an IVIS Spectrum which has extended our capability into non-invasive in-vivo imaging in both bio luminescence and fluorescence modes. Widefield epifluorescence and brightfield imaging of histopathology is also available

We ask that new users to contact Rob Shaw before they commence any staining protocols.

To request training go to the MISBU facility on Stratocore start page https://ppms.eu/bham/start/ 

Zeiss 780 Live Cell confocal...

The latest addition is an LSM780 and is the first of its type in the UK. The 780 has high sensitivity and is equipped with a 32 array and 2 galanium arsenide phosphide (GaAsP) detectors giving much higher quantum efficiency of normal PMT’s. The system has the same laser lines as the LSM 510 Meta confocal (405, 458, 488, 514, 561, 594 and 633nm) and can separate dyes efficiently via a spectral AOTF allowing a maximum of 8 channels in sequential imaging mode. Over 100 dye configurations are available in the Zen 2010 confocal software. It can be used for live cell confocal imaging having an inverted heated stage and incubation chamber

Microscope stand

  • Axio Observer.Z1 inverted microscope
  • Interchangeable stage inserts for slides, 30mm dishes and multiwall plates
  • Fully motorised stage in x, y, & z, with joystick x/y control
  • LCD docking station with integrated epifluorescence, course / fine focus controls
  • Pecon incubation system incorporating ducted air, heated stage and CO2 induction for live cell imaging
  • Sample illumination: 
    o  HAL 100W tungsten transmitted light
    o  100W HXP120 long life mercury epifluorescence burner

Epifluorescence filter sets

Filter set

Ex BP nm

Em BP nm

Fs49 wf

365

445/50

Fs38 wf

470/40

525/50

Fs43 wf

545/25

635/70

Optics

Objective lens

Magnification / NA / Immersion

Contrast Method

C-ACHROMAT

10x / 0.45 / W

 

LD LCI Plan-NEOFLUAR

25x / 0.8 / H20 or Oil

DIC II

C-ACHROMATIC

40x / 1.2 / W

DIC III

C-ACHROMAT

63x / 1.2 / W

DIC III

Plan-ACHROMAT

100x / 1.4 / Oil

DIC III

Confocal lasers

  • Lasos 30mW Diode 405nm
  • Lasos 25mW LGN3001 multiline Argon, 458, 488, 514nm
  • Lasos 20mW (DPSS 561)   561nm
  • Lasos 2mW   HeNe 594nm
  • Lasos 5mW   HeNe 633nm

Software and Network Drives

  • Zen 2010 software, with up to 8 confocal fluorescence channels plus transmitted light detection.
  • Z stack with orthogonal view, 2D projection and 3D rendering
  • Data acquisition in x, y, z, & t
  • Multi position referencing
  • Tile scanning and image stitching of large samples
  • Bleaching (FRAP)
  • Networked to the Technology Hub (data) server for optimised image capture and offline retrieval.

Location: IBR level 4, room 411

Accounts / access / training:

Stratocore; https://ppms.eu/bham/login/?pf=4

Carl Zeiss; http://microscopy.zeiss.com/microscopy/en_gb/home.html

Zeiss LSM 510 Meta confocal...

The LSM 510 Meta confocal is based on an upright microscope with an x,y,z motorised stage. Tiling of images is possible as well x, y, z, & t data acquisition. Available laser lines are 405, 458, 488, 514, 561, 594 and 633. The Meta head is capable of spectrally separating dyes thus increasing the number of signals/channels available.

Microscope stand

  • Upright Axio Imager Z2
  • Motorised stage in x,y,& z
  • Sample illumination 
    o  HAL 100W tungsten transmitted light 
    o  100W HB0 short arc mercury epifluorescence burner

Epifluorescence filter sets

Filter set

Ex BP nm

Em BP nm

Fluorophores

49

365

445/50

DAPI, Hoechst

10

450-490

515/565

FITC, Alexa 488, GFP

20

546/12

575-640

Cy3, TRITC DsRed

26

575-625

600-710

Cy5, Alexa 633,647

Optics

Objective lens

Magnification / NA / Immersion

Contrast Method

C-ACHROMAT

10x / 0.45 / W

 

LCI Plan-NEOFLUAR

25x / 0.8 / H20 or Oil

DIC

C-ACHROMATIC

40x / 1.2 / W

 

C-ACHROMAT

63x / 1.2 / W

 

Plan-ACHROMAT

100x / 1.4 / Oil

DIC

Confocal lasers

  • Lasos 30mW Diode (405-30), 405nm
  • Lasos 30mM multiline Argon/2, 458nm, 477nm, 488nm, 514nm
  • Lasos 15mW DPSS (561-10), 561nm
  • Lasos 2mW HeNe, 594nm
  • Lasos 5mW HeNe, 633nm

Software and Network Drives

  • LSM 510, 4 fixed confocal fluorescence channels including 1 variable Meta channel with spectral separation and detection, plus transmitted light detection.
  • x, y, z, & t acquisition, z-stack with orthogonal view, 2D and 3D reconstruction
  • Tile scanning of large images
  • Networked to the Technology Hub (data) server for optimised image capture and offline retrieval.

Location: IBR level 4, room 411

Accounts / access / training:

Stratocore; https://ppms.eu/bham/login/?pf=4

Carl Zeiss; http://microscopy.zeiss.com/microscopy/en_gb/home.html

Zeiss LSM 510 UV-confocal...

The LSM 510-UV confocal has 351, 361, 458, 488, 543 and 633 laser illumination and is based on an inverted microscope that is capable of taking high resolution images in x, y, z & t. The stage is motorised for compiling z stacks producing 2D and 3D images. Certain stage inserts can be heated for live cell work, more…….

Microscope stand

  • Axiovert 100M inverted
  • Stage inserts for multiwall plates and a thermo regulated heated block for live cell imaging (30mm imaging dishes)
  • Motorised stage for z-stack and 2D / 3D reconstruction
  • Sample illumination
    o   HAL 100W tungsten transmitted light
    o   100W HB0 short arc mercury epifluorescence burner
    o   Motorised slider with blue, green and red emission filters

Optics

Objective lens

Magnification / NA / Immersion

Contrast Method

ACHROMAT

10x / 0.45 / W

Phase

LCI Plan-NEOFLUAR

25x / 0.8 / W

DIC

C-ACHROMATIC

40x / 1.2 / W

 

C-ACHROMAT

63x / 1.2 / W

 

Plan-ACHROMAT

100x / 1.4 / Oil

DIC

Confocal Lasers

  • Coherent Enterprise II 80mW UV multiline, 351nm, 361nm
  • Lasos 25mW multiline Argon, 458nm, 488nm
  • Lasos 1mW HeNe, 561nm
  • Lasos 5mW HeNe, 633nm

Software and Network Drives

  • LSM 510 with 4 confocal fluorescence channels plus transmitted light detection.
  • Z stack with orthogonal view, 2D projection and 3D reconstruction
  • Networked to the Technology Hub (data) server for optimised image capture and offline retrieval.

Location: IBR level 4, room 411

Accounts / access / training

Stratocore; https://ppms.eu/bham/login/?pf=4

Carl Zeiss; http://microscopy.zeiss.com/microscopy/en_gb/home.html

Leica LSI Macro Zoom - True Confocal Scanner...

The Leica TCS LSI macro confocal zoom confocal offers high resolution with true confocal scanning incorporating spectral technology plus a large 16mm field of view for small in vivo imaging. The automated freely adjustable optical zoom system allows seamless magnification adjustment, overcoming the limitations of fixed objectives but retaining high resolution imaging of the model organism. More……macro confocal zoom confocal offers high resolution with true confocal scanning incorporating spectral technology plus a large 16mm field of view for small in vivo imaging. The automated freely adjustable optical zoom system allows seamless magnification adjustment, overcoming the limitations of fixed objectives but retaining high resolution imaging of the model organism.

  • Rack mounted Z16a scanner and integral zoom optics for precise motor driven focussing
  • Spectral AOTF with dye separation, and sequential scanning.
  • Imaging chamber with swing open doors, adaptable for full environmental control
  • Multiple stage inserts for slides, dishes and multiwell plates.
  • Additional epifluoescence imaging (100W long life mercury short arc burner)

Epifluorescence filter sets

Filter block

Ex BP nm

Em BP nm

Fluorophores

A4

365

445/50

DAPI, Hoechst

I3

450-490

515/565

FITC, Alexa 488, GFP

N2

546/12

575-640

Cy3, TRITC DsRed

L5

575-625

600-710

Cy5, Alexa 633,660,647

Optics

Objective lens

Magnification / NA / working distance

Contrast Method

Plan Apo

2x / 0.03 / 39mm

Phase

Plan Apo

5x / 0.08 LWD

Phase

 

 

Optical zoom

 Factor 0.5x to 9.29x

 

Confocal Lasers

  • Lasos Diode 405nm
  • Lasos Diode 488nm
  • Lasos Diode 531nm
  • Lasos Diode 647nm

Software and Network Drives

  • LAS AF software with true spectral AOTF serving 4 confocal fluorescence channels plus transmitted light detection.
  • Z stack with 2D projection
  • Tile scanning and image stitching of large samples
  • Leica desktop potentiometers with LCD docking station and ergonomic x/y/z control
  • Epifluorescence image capture via Leica DFC 310 FX digital firewire camera 1392x1040
  • Networked to the Technology Hub (data) server for optimised image capture and offline retrieval

Location: IBR west level 1, room WX1.61

Accounts / access / training

Stratocore; https://ppms.eu/bham/login/?pf=4

www.leicamicrosystems.com

http://www.leica-microsystems.com/products/confocalmicroscopes/details/product/leica-tcs-lsi/downloads/

http://www.leica-microsystems.com/fileadmin/downloads/Leica%20TCS%20LSI/Brochures/Leica%20HCS%20LSI-Brochure_EN.pdf

Leica DM6000B widefield epifluorescence / brightfield microscope...

The Leica DM6000 is multimodality with a 4 channel epifluorescence and bright field system incorporating Leica Application Suite Advanced Fluorescence(LAS AF) software. Rapid tile scanning of large images is achievable with multi position settings and autofocus. High resolution colour imaging of histopathology can also be achieved using the Q Imaging colour camera and Q Capture software. More………

Microscope stand

  • DM6000B upright with LCD touch panel
  • Fully motorised stage in x, y, & z with ergonomic joystick
  • Sample illumination 
    o  100W tungsten transmitted light
    o  100W short arc epifluorescence mercury burner 
    o  Motorised dichroic filter turret

Epifluorescence filter sets 

Filter cube

Excitation nm

Emission

nm

A4

360

Blue 470

L5

480

Green 527

N3

546

Red 600

Y5

620

Far Red 700

Optics

Objective lens

Magnification / NA / Immersion

Contrast Method

HC Plan Apo

10x / 0.45 / Dry

Phase 1

HC Plan Apo

20x / 0.7 / Dry

 

Phase 3

Plan Apo

40x / 1.25 / Oil

 

 

Cameras

  • Leica DFL350 FX firewire camera
  • Q Imaging Micropublisher 5.0 RTV colour camera
  • Software features and Network Drives
  • Leica LAS AF software, 4 channel widefield fluorescence and brightfield 
    o  Multipoint imaging with autofocus at each x/y point
    o  Tile scanning and image stitching of large samples
  •  Q capture imaging software for colour imaging of histopathology
  • Networked to the Technology Hub (data) server for optimised image capture and offline retrieval.

Location: IBR level 4, room 411

Accounts / access / training

Stratocore; https://ppms.eu/bham/login/?pf=4

Leica microsystems; http://www.leica-microsystems.com/home/

Leica DMI6000 time-lapse widefield epifluorescence / brightfield imager...

Based in the Centre for Translational Inflammation Research (CTIR) the Leica the DMI6000 provides 3 colour epi-fluorescence and time-lapse imaging using the Leica Application Suite Advanced Fluorescence(LAS AF) software. More…..

Microscope stand

  • DMI6000 upright with LCD touch panel
  •  Motorised stage in x, y, & z with ergonomic joystick
  • Sample illumination 
    o  100W tungsten brightfield 
    o  100W HXP120 long life mercury epifluorescence burner 
    o  Motorised emission filter turret, (L5 green, N3 Red, A4 Blue).

Optics

Objective lens

Magnification / NA / Immersion

Contrast Method

HCX Pl Fluotar

20x / 0.4 / Corr Dry

Phase 1

HCX Pl Fluotar

40x / 0.6 / Corr Dry

Phase 2

HCX Pl Fluotar

63x / 0.7 / Corr Dry

Phase 2

Camera

  • Leica DFL350 FX firewire camera

Software features and Network Drives

  • Leica LAS AF software, 3 channel widefield fluorescence and brightfield
  • Time-lapse mode
  • Networked to the Technology Hub (data) server for optimised image capture and offline retrieval.

Location: Confocal Microscopy, Centre for Translational Inflammation Research, QEH.

Accounts / access / training:

Stratocore; https://ppms.eu/bham/login/?pf=4

Leica microsystems; http://www.leica-microsystems.com/home/

IVIS Spectrum Whole Animal Imager...

The IVIS Spectrum uses Xenogen’s novel patented optical imaging technology to facilitate non-invasive longitudinal monitoring of disease progression, cell trafficking and gene expression patterns in living animals. The IVIS Spectrum is a versatile and advanced in-vivo imaging system available on the market today. An optimized set of high efficiency filters and spectral un-mixing algorithms lets you take full advantage of bioluminescent and fluorescent reporters across the blue to near infrared wavelength region. It also offers single-view 3D tomography for both fluorescent and bioluminescent reporters that can be analysed in an anatomical context using a Digital Mouse Atlas.

For advanced fluorescence imaging, the IVIS Spectrum has the capability to use either trans-illumination (from the bottom) or epi-illumination (from the top) to illuminate in vivo fluorescent sources. 3D diffuse fluorescence tomography can be performed to determine source localization and concentration using the combination of structured light and trans illumination fluorescent images. The instrument is equipped with 10 narrow band excitation filters (30nm bandwidth) and 18 narrow band emission filters (20nm bandwidth) that assist in significantly reducing autofluorescence by the spectral scanning of filters and the use of spectral unmixing algorithms. In addition, the spectral unmixing tools allow the researcher to separate signals from multiple fluorescent reporters within the same animal.

Main Features

Imaging Chamber

  • Different stage heights allow adjustable field of view, from well plates (3.9 cm field of view = highest resolution at 20µm) to high-throughput animal imaging (5 mice)
  • Integrated isofluorane anaesthesia system
  • LED photographic overlay image
  • Heated stage with for animal optimisation
  • Safety electromagnetic door latch
  • Motorised stage, filter wheels, lens positions, and f-stop controls
  • Laser illuminated stage layout for mouse positioning and alignment
  • Multifunctional filter wheel for bioluminescence, fluorescence and trans-illumination imaging

CCD Camera and Optics

  • Back-illuminated 16 bit CCD is thermoelectrically cooled to -90OC for reduce noise
  • CCD Format 2048 x 2048 pixels
  • High quantum efficiency over the entire visible to near-infrared spectrum
  • f-stop settings from f/1– f/8, 150mm imaging lens
  • CCD Size 26 x 26 mm
  • Effective CCD Format 1920 x 1920 pixels
  • Quantum Efficiency ~85% 400-700nm
  • Dark Current <100 electrons/s/cm2 (-90° C)
  • Minimum Detectable Luminance <70 photons/s/sr/cm2
  • CCD Temperature Nominal -90°

Access

For all bookings email; biomedicalservices@bham.ac.uk

Contacts:

Perkin Elmer home: http://www.perkinelmer.co.uk/default.xhtml

IVIS Spectrum: http://www.perkinelmer.co.uk/Catalog/Product/ID/IVISSPE

http://www.perkinelmer.co.uk/CMSResources/Images/44-135294BRO_SPEC-BR-01-6708.pdf