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Confocal microscopy equipment and specifications

Below is a description of the major items of equipment available. We strongly encourage you to seek our advice in the early planning of your project.

To use our equipment you will need to register your project and undergo training.

Confocal microscopy registration

For advice contact:
Email andrew.mcnaughton@otago.ac.nz

Nikon A1+ Inverted Confocal Laser Scanning Microscope

Nikon A1+ microscope imageAn inverted system, the confocal is suitable for small petri dishes, well plates and conventional slide-mounted specimens. It has laser excitation at 405nm, 488nm, 561nm and 640nm, plus DIC and an RGB camera, suitable for recording conventional fluorescence images.

Specifications

  • Model: Nikon A1+
  • Microscope Frame: Nikon TiE
  • Manufacturer: Kurobane Nikon Co., Ltd. 1434, Kurobanemuko-machi, Otawara, Tochigi 324-0241, Japan

Objectives

  • 10× Plan
  • 20× PlanApo
  • 40× PlanApo
  • 60× PlanApo (oil)

Camera

Nikon DSiR2 colour 16MP

Laser (solid state) excitation lines

  • 405nm
  • 488nm
  • 561nm
  • 640nm

Fluorescence filters

  • UV excitation (emission filters 450 / 50)
  • Blue excitation (emission filters 525 / 50)
  • Green excitation (emission filters 595 / 50)
  • Red excitation (emission filters 700 / 75)

Software

Nikon Elements C, running on Windows 7

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Nikon A1R Multi-Photon Confocal Laser Scanning Microscope

Nikon A1R microscope imageFeaturing state of the art optics and detectors, the A1R has the potential to capture both fleeting physiological events and probe deep into both fixed and in vivo tissues using its multi-photon features. Highly customisable and comprehensive software allows each experiment to be fine tuned to the user’s requirements.

This microscope requires careful operation and training is essential.

Specifications

  • Model: Nikon A1R MP
  • Microscope Frame: Nikon Eclipse
  • Manufacturer: Kurobane Nikon Co., Ltd. 1434, Kurobanemuko-machi, Otawara, Tochigi 324-0241, Japan

Objectives

  • 4× Plan 4× 0.1 NA
  • 25× Apo LWD 1.0 NA (water immersion)
  • 40× Plan Fluor 0.75 NA w/d 0.66mm

Laser (solid state) excitation lines

  • 405nm
  • 488nm
  • 561nm
  • 635nm

Fluorescence filters

  • UV excitation (emission filters 450 / 50)
  • Blue excitation (emission filters 525 / 50)
  • Green excitation (emission filters 595 / 50)
  • Red excitation (emission filters 700 / 75)

Software

Nikon Elements C, running on Windows 7

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Olympus AX70 Light Microscope

Olympus AX 70 microscope imageA standard research level light microscope with brightfield, phase contrast, dark field, and fluoescence. The AX70 has an upright configuration and is suited to fixed material on standard microscope slides.

Fluorescence ranges from uv, blue, green and red excitation using a xenon burner. Epi-fluorescence is also available and is useful for such techniques as silver enhanced immuno-gold labelling.

Specifications

  • Model: Olympus AX70
  • Manufacturer: Olympus Corporation, Shinjuku Monolith, 2-3-1 Nishi-Shinjuku, Shinjuku-ku, Tokyo 163-0914, Japan
  • Camera: QImaging Micropublisher 5.0 RTV. 2560×1920 pixels, 3.4µm pixel size. Diagnostic Instruments, 19535 56th Avenue, Suite 101, Surrey, BC, Canada V3S 6K3

Objectives

  • 1.25× PlanApo 0.04NA
  • 4× UPlanFl 0.13NA
  • 10× UPlanFl 0.3NA
  • 20× UPlanSApo 0.75NA
  • 40× UPlanSApo 0.95NA
  • 60× PlanApo 1.40NA (oil immersion)

Fluorescence filters

  • Blue excitation bandpass 460-490nm / longpass emission filter 515nm
  • Green excitation bandpass 520-550nm / longpass emission filter 580nm
  • Red excitation bandpass 540-580nm / longpass emission filter 610nm

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Olympus BX51 Light Microscope with StereoInvestigator

Olympus BX51 microscope with stereoinvestigator software imageA sophisticated stereology software package has been integrated onto an Olympus BX51 light microscope with motorised xy stage and z stepper motor. This system is capable of complex stereological analysis of your light microscopy images, including those collected on other light and the confocal microscopes. This instrument is located in room 107 on the first floor of the Scott Building.

StereoInvestigator is a powerful stereology software package which can take you from simple point counting to exceptional analysis of your images. It can be used in a two dimensional mode to estimate the frequency of say cell types, collating them into categories and performing appropriate statistical analysis. It can also be used in three dimensional analysis, focusing through a thick section and marking the change in cell frequency through the planes of focus.

It is strongly advised you obtain advice in advance of beginning your investigation.

StereoInvestigator version 2017 overview (YouTube)

StereoInvestigator version 2018 information (MBF Science website)

Specifications

  • Model: Olympus BX51
  • Microscope Manufacturer: Olympus Corporation, Shinjuku Monolith, 2-3-1 Nishi-Shinjuku, Shinjuku-ku, Tokyo 163-0914, Japan
  • Camera Manufacturer: Ximea xiD, XIMEA GmbH, Hansestraße 81, 48165 Münster, Germany
  • Camera Specifications: Ximea xiD series (MD061CU-SY), 6 Megapixel Color CCD (Sony ICX694) Camera pixel size 4.54 μm

Objectives

  • 4× 0.16 NA, PlanApo
  • 10× 0.4 NA, UPlanApo
  • 20× 0.7 NA, UPlanApo
  • 20× 0.5 NA, Phase 1, UPlanFl
  • 40× 0.7 NA, Phase 2, UPlanFl
  • 100× Plan 1.25 NA (oil immersion)

Fluorescence filters

  • UV excitation bandpass 330-385nm / longpass emission filter 420nm (WU filter cube)
  • Blue/Violet excitation bandpass 400-440nm / longpass emission filter 475nm (WBV filter cube)
  • Blue excitation bandpass 460-490nm / longpass emission filter 515nm (WIB filter cube)
  • Green excitation bandpass 520-550nm / longpass emission filter 580nm (WIG filter cube)
  • Yellow excitation bandpass 540-580nm / longpass emission filter 610nm (WIG filter cube)

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Olympus BX61 Montaging Light Microscope

Olympus BX61 Montaging Light microscope image

Specifications

  • Model: Olympus BX61
  • Manufacturer: Olympus Corporation, Shinjuku Monolith, 2-3-1 Nishi-Shinjuku, Shinjuku-ku, Tokyo 163-0914, Japan
  • Camera: QImaging Micropublisher 5.0 RTV. 2560×1920 pixels, 3.4µm pixel size. Diagnostic Instruments, 19535 56th Avenue, Suite 101, Surrey, BC, Canada V3S 6K3

Objectives

  • 2× PlanApoN 0.08NA
  • 4× UPlanSApo 0.16NA
  • 10× UPlanSApo 0.4NA
  • 20× UPlanSApo 0.75NA
  • 40× UPlanSApo 0.95NA
  • 100× UPlanSApo 1.40NA (oil immersion)

Fluorescence filters

  • UV excitation bandpass 330-385nm / longpass emission filter 420nm (WU filter cube)
  • Blue excitation bandpass 460-490nm / bandpass emission filter 515-550nm (WIBA filter cube)
  • Green excitation bandpass 520-550nm / longpass emission filter 580nm (WIG filter cube)

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Skyscan µCT Scanner

Skyscan micro computed tomography scanner imageA high resolution µCT scanner, the Skyscan 1172 uses X-ray tomography to generate 3D data sets of specimens measuring up to 55×55×55mm. Resolutions of 2µm are possible and a wide range of specimens can be examined, from food to teeth. Soft tissues usually require a contrasting agent to be used.

Large amounts of data (4–8 GB per sample) are produced and if volume rendering is required, a large amount of computing capacity too. Ask about the best approach while planning your investigation.

Specifications

  • Model: Skyscan 1172
  • Manufacturer: Bruker-MicroCT, Kartuizersweg. 3B, 2550, Kontich, Belgium
  • Camera: 10 megapixel, (4000×2300), 12 bit digital, cooled CDD coupled by fibre optic to scintillator (50mm field of view)

Pixel size and log file

This varies depending upon the settings used to obtain images. It's the number you need to know if you want to make spatially calibrated measurements of your data, otherwise they are just so many pixels in an area, rather than say square millimetres. Check your log file (which is automatically generated with your data) line 25 ‘Image pixel size (µm) = xx’ shows this parameter’s value for your data.

When your raw data is processed to produce the slice data used in analysis, a second log file is generated. This appends processing parameters to the scan parameters used to acquire the image, it’s found among the processed slice images. The easiest way to find it is to sort your folder by size as it's very small compared to the images. The details after line 48 ‘[Reconstruction]' relate to generating the slice images, details before this line relate to image acquisition in the scanner.

We strongly recommend you read the log file, it is very informative.

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Zeiss LSM 710 Upright Confocal Laser Scanning Microscope

Zeiss LSM 710 microscope imageUpright configuration on an Axio Imager Z2 frame with 405 diode laser, argon laser, green HeNe and red HeNe laser, phase objectives and mercury burner.

This configuration is best suited for fixed material on standard microscope slides. It can unmix overlapping spectra that are difficult to separate on the Zeiss 510 confocals. It can also perform tile scans over the z axis which is useful for large samples.

Specifications

  • Model: Zeiss LSM 710 confocal laser scanning microscope
  • Microscope Frame: Zeiss AxioImager Z2, upright microscope with motorized stage
  • Manufacturer: Carl Zeiss Microscopy GmbH, Carl Zeiss Promenade 10, 07745, Jena, Germany

Objectives

  • 5× PlanNeoFluar 0.16NA
  • 10× PlanApo 0.45NA
  • 20× PlanApo 0.8NA
  • 40× PlanApo 1.30NA (oil immersion)
  • 63× PlanApo 1.40NA (oil immersion)

Laser excitation lines

  • 405nm diode laser
  • 458, 488 and 514nm argon laser
  • 543nm helium neon (HeNe) laser
  • 633nm helium neon laser (HeNe) laser

Fluorescence filters

  • UV excitation (filter set 49)
  • Blue excitation (filter set 38)
  • Green excitation (filter set 43)

Software

Zeiss ZEN 2009, running on Windows Vista

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