Light Microscopy

CONTENTS


Nuance Multi-Spectral Imaging System

Nuance

Location
Room E401, Anderson Stuart Building

Equipment

The Nikon Eclipse E800 Macro-Imaging Microscope has been upgraded and renamed to the Nuance Multi-Spectral Imaging System. The old Leica DFC500 camera and LAS software has been replaced with the Nuance camera and multi-spectral acquisition software for both bright-field and fluorescence imaging.

This Nuance camera allows multi-spectral data acquisition using a liquid crystal tunable filter that provides fast, accurate spectral imaging from 420 to 720 nm (VIS) and 500 to 950 nm (NIR), covering a range of available dyes and fluorophores. This multi-spectral imaging technology can therefore be used to acquire images at many wavelengths and thus can better determine fluorophore/stain distribution in cells or tissue. The Nuance system can even separate spatially overlapping chromogens and help to remove autofluorescence from tissues.

Nuance Multi-Spectral Imaging System
Wavelength ranges:

  • FX: 420 to 720 nm (regular microscopy)
  • EX: 450 to 950 nm (in vivo imaging and validation)

Features:

  • Electronically tunable bandpass filter
  • Excellent image quality – no pixel shift, no moving parts
  • Equivalent to approx. 150 filters in a single filter wheel!
  • Rapid tuneability
  • Excellent spectral quality
  • Software-selectable bandpass: 20 nm or 40 nm
  • Minimum step size down to 1 nm

Funds for this upgrade were provided by a Bosch/RIBG 2014 small equipment grant.


Olympus MBF/Stereo Investigator Transmitted Light Microscope

Location
Room S448, Anderson Stuart Building

StereoInvestigator

The old Leica DMRBE upright light microscope stand for the Sterology Microscope has been replaced in 2013 with a new Olympus BX53 Stand. It then been renamed the Olympus/MBF Stereo Investigator Transmitted Light Microscope with the motorised xyz-stage and MBF Stereo Investigator software for design based Stereology, Image Analysis, and 2D Anatomical Mapping (purchased from SciTech).

The Virtual Slice module that automatically creates a montage by collecting a series of contiguous images from a tissue specimen. The software aligns, stitches and blends images into a seamless montage. This microscope is set up for bright-field imaging and
enables fast navigation to any region of the slide.

Funding for the purchase of this upgrade was provided by NHMRC Equipment Grant 2008 and Rebecca L. Cooper Medical Foundation Grant 2009.

llumination sources

  • 12V/ 100W Halogen lamp

Objectives

  • Plan fluo 5x dry/ NA 0.12/ WD 0.12mm
  • Plan apo 20x dry / NA 0.50/ WD 0.15mm
  • Plan apo 40x dry/ NA 0.75/ WD 0.08mm
  • Plan apo 100x oil/ NA 1.40/ WD 0.09mm

Zeiss "Deconvolution" Microscope

Bringing microscopes up-to-date: The old AxioPlan microscope stand for the Zeiss "deconvolution" microscope purchased in 2001 that was showing signs of wear and tear was replaced in March 2012 with a new Zeiss AxioImager Stand. Funding for this upgrade was provided by RIBG funding in 2012. This latter stand has a motorized transmitted-light beam path and focus drive and can be operated via a touch-sensitive TFT display, by pressing buttons directly on the microscope or via the ZEN (ZEISS Efficient Navigation) software. This new Axio Imager.M2 offers a higher level of convenience and performance over the old stand. The system is ideal for imaging specimens using different transmitted-light contrasting and fluorescence techniques with reproducible settings.
In 2012, a new imaging system, named ApoTome.2, was purchased from Carl Zeiss Australia and installed onto the Zeiss "Deconvolution" Microscope. The ApoTome.2 creates optical sections of the fluorescent samples by using structured illumination. This imaging system recognizes the magnification and moves the appropriate grid into the beampath. The system then calculates the optical section from three images with different grid positions without time lag. It’s a totally reliable way to prevent scattered out-of-focus light, even in thicker specimens. Funding for this equipment was supported by the Rebecca L. Cooper Medical Research Foundation.

Location
Room S443, Anderson Stuart Building

Zeiss Decon 2013

Zeiss "Deconvolution" Microscope System with the new ApoTome.2

The facility
The system comprises:

  • Zeiss Axio Imager.M2 upright microscope, automated
  • ZEN - ZEISS Efficient Navigation software (includes Deconvolution and 4D rendering modules)
  • Zeiss AxioCam HRm digital monochrome CCD camera
  • Zeiss AxioCam HRc digital colour CCD camera
  • Zeiss ApoTome.2

Zeiss Axio Imager.M2 Upright microscope
Illumination sources

  • 12V/ 100W HLX Xenophot 64625 Halogen lamp
  • HBO 103W Hg short arc lamp

NEW reflected light filters

  • DAPI - 365, 395LP, 420LP
  • Alexa 488 - 480/40, 505LP, 535/50
  • Texas Red - 560/55, 595LP, 645/75
  • CY3 - 545/30, 570LP, 610/75
  • CY5 - 620/60, 660LP, 700/75

Objectives

  • Plan Apochromat 5x dry/ NA 0.16
  • Plan Neofluar 10x dry/ NA 0.3/ WD 5.6mm
  • Plan Neofluar 20x dry/ NA 0.50/ WD 2.0mm
  • Plan Neofluar 40x oil/ NA1.3/ WD 0.17mm
  • Plan Apochromat 63x oil/NA 1.4/ WD 0.09mm

ZEN - ZEISS Efficient Navigation software

Acquisition

  • Multiple-channel images
  • Image z-stacks

Processing

  • Processing of 14-bit image (brightness, contrast, gamma)
  • Image deconvolution
  • 3D-rendering (x, y, z) and 4D-rendering (x, y, z, t) with Inside 4D module

File saving

  • Network
  • CD or DVD+R/ RW
  • USB devices