Light Microscopy

CONTENTS


Nikon Eclipse E800 Macro-Imaging Microscope

Nikon

Location
Room S448, Anderson Stuart Building

Equipment

  • Leica DFC500 colour camera and LAS imaging software (including additional Overlay and Multifocus modules). Supported by Rebecca L. Cooper Medical Foundation Grant 2008

Nikon Eclipse E800 Upright Microscope
Illumination sources

  • 12V/ 100W Halogen lamp
  • HBO 103W/2 Hg short arc lamp

Reflected light filters

  • UV (Ex 330-380, DM 400, BA 420)
  • Blue (Ex 450-490, DM 505, BA 520)
  • Green (Ex 510-560, DM 575, BA 590)

Objectives

  • MACRO Plan 0.5x/ NA 0.025/ WD 7.0mm
  • PlanApo 4x/ NA 0.2/ WD 15.7mm
  • PlanApo 10x/ NA 0.45 DIC L/ WD 4.0mm
  • PlanApo 20x/ NA 0.75 DIC M/ WD 1.0mm
  • PlanApo 40x corr/ NA 0.95 DIC M/ WD 0.14mm
  • PlanApo 100x/ NA 1.40 DIC H/ WD 0.13mm

Leica Stereology Microscope

The following facilities are availble:

Location
Room S448, Anderson Stuart Building

Upright

Leica DMRBE Upright Microscope
The Leica DMRBE upright light microscope that was part of the old Leica TSCNT confocal microscope has been renamed the Leica Stereology Microscope with the addition of a motorised xyz-stage and Stereo Investigator software for Design based Stereology, Image Analysis, and 2D Anatomical Mapping (purchased from SciTech).

The Virtual Slice module allows you to simply create and analyse high-resolution image montages. The microscope is set-up for transmitted light and basic fluorescence microscopy which includes standard DAPI, FITC and Rhodamine filters sets.

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

Images from top to bottom: Whole virtual slice, Virtual slice software, Needle track

llumination sources

  • 12V/ 100W HLX Xenophot 64625 Halogen lamp
  • HBO 50W Hg short arc lamp

Reflected light filters

  • UV (A 51382) - BP 340-380, DM 400, SF LP 425
  • Blue (I3 513828) - BP 450-490, DM 510, SF LP 515
  • Green (N2.1 513832) - BP 515-560, DM 580, SF LP 590
  • 1 blank filter space

Objectives

  • Plan fluotar 5x dry/ NA 0.12/ WD 0.12mm
  • Plan fluotar 16x imm ph2/ NA 0.50/ WD 0.15mm
  • Plan fluotar 40x oil/ NA 1.00-0.50/ WD 0.08mm
  • Plan apo 63x oil/ NA 1.32-0.6/ WD 0.07mm
  • Plan apo 100x oil/ NA 1.40-0.7/ WD 0.09mm
  • Plan apo 100x oil ph3/ NA 1.40/ WD 0.09mm
  • Plan apo 63x water corr/ NA 1.20/ WD 0.22mm

Leica DMIRBE Inverted Microscope
llumination sources

  • 12V/ 100W HLX Xenophot 64625 Halogen lamp
  • HBO 50W Hg short arc lamp

Reflected light filters

  • UV (A 51382) - BP 340-380, DM 400, SF LP 425
  • Blue (I3 513828) - BP 450-490, DM 510, SF LP 515
  • Green (N2.1 513832) - BP 515-560, DM 580, SF LP 590
  • 1 blank filter space

Objectives

  • Neo plan 5x ph0/ NA 0.12/ WD 14.0mm
  • Plan fluotar 16x imm/ NA 0.50/ WD 0.15mm
  • Plan fluotar 40x oil ph3/ NA 1.00/ WD 0.08mm
  • Plan apo 63x oil ph3/ NA 1.32/ WD 0.07mm

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