Equipment Information

Optical Microscopy
Confocal Microscopy
Laser Capture Microdissection
Live Cell Imaging

Digital Imaging and Image Analysis
Scanners
Printing
Software

Optical Microscopy

Location Research Building CHW, Level 3 SPOT Room 43 4876
Contact Laurence Cantrill Lab 1 Level 3, Phone 9845 3087

Olympus BX50 microscope (SPOT) with:

• Bright field
• Fluorescence, HBO 100W/2 Hg short arc lamp
• Polarised light
• Phase
• DIC (10x, 20x, 40x, 100x)

• Jenoptik ProgRes CF Scan digital camera with ProgRes V2.5 software

Leica DMLA Automated Microscope (DC500) with:

• Bright field
• Fluorescence HBO 100W Hg short arc lamp

CPF, YFP and far red (CY5) filters can also be supplied. Please check with the microscopist.

• Objectives

• QICAM fast 1394 colour 12 bit camera with Q Capture Software/ Bioquant Nova Prime data capture software
• Leica DC500 colour camera with Leica IM50 image capture software
• File saving, CD/DVD burner, USB port

Leica MZ8 Dissecting Microscope

• Brightfield only (0.63- 5x magnification)
• Leica DC500 colour camera with Leica IM50 image capture software or QICAM fast 1394 colour 12 bit camera with Q Capture Software
• Schott KL 1500 LCD cold light source with goose neck light guides for overhead illumination

Confocal Microscopy

Leica TCS SP2

Location: Research Building CHW, Level 4 Confocal Room 44 4873
Contact: Laurence Cantrill, Lab1, Level 3 Office 9845 3087

The Confocal Microscope was purchased in 2001 by the Oncology Research Unit at the Children's Hospital at Westmead. The microscope was purchased with money raised mostly from donations to the Children's Hospital. The system runs on a Windows XP workstation with dual high resolution 21" screens.

The confocal facility consists of 2 microscopes

DMRBE (Upright microscope)
DMIRBE (Inverted microscope) with Live Cell Imaging Camera

	Upright		Inverted
Illumination	12V/ 100W HLX Xenophot 64625 Halogen lamp, EL6000 long life fluorescence system		12V/ 100W HLX Xenophot 64625 Halogen lamp, HBO 50W Hg short arc lamp
Epifluorescence Excitation filters	A, UV BP 340-380nm (DAPI) I3, blue BP 450-490nm (FITC, Alexa 488) N2.1, green BP 515-560 (Cy3, TRITC, Alexa 543, 555, 568, TR, PI) CGFP, violet/blue BP 436/20 YFP, blue/green BP 500/20 Y5, red BP 590-650 (CY5, Alexa 633, DRAQ5)		A, UV BP 340-380nm (DAPI) I3, blue BP 450-490nm (FITC, Alexa 488) N2.1, green BP 515-560 (Cy3, TRITC, Alexa 543, 555, 568, TR, PI) Y5, red BP 590-650 (CY5, Alexa 633, DRAQ5)
Epifluorescence Emission filters	A, violet LP 425nm- (DAPI) I3, green LP 515- (FITC, Alexa 488) N2.1, red LP 590-(Cy3, TRITC, Alexa 543, 555, 568, TR, PI) CGFP, blue BP 480/40YFP, green/yellow BP 535/30 Y5, far red BP 675-735 (CY5, Alexa 633, DRAQ5)		A, violet LP 425nm- (DAPI) I3, green LP 515- (FITC, Alexa 488) N2.1, red LP 590-(Cy3, TRITC, Alexa 543, 555, 568, TR, PI) Y5, far red BP 675-735 (CY5, Alexa 633, DRAQ5)
Laser Illumination	ArKr (458nm, 476nm, 488nm, 514nm lines), Green HeNe (543nm line), Red HeNe (633nm line). Class 3B lasers.
Laser Emission	Acousto-optical tuneable filters (AOTF)
Detectors	4 Fluorescence PMT+ 1 transmitted detector (for BF, Phase (16x, 25x, 40x, 63x, 100x), DIC (40x, 63x, 100x))
Stage control	Electronic microscope XY stage, Z galvo stage		Electronic microscope stage, Z wide stage
Objective Lenses		Lens/NA	Type	Working Distance
	HC PL Fluotar	4x/0.1	Dry	26.2mm
	HC PL Fluotar	5x/0.15	Dry	12mm
	Plan Fluotar (PH2)	16x/0.5	IMM (Water, Glycerine, Oil)	150μm
	Plan Fluotar (PH2)	25x/0.75	Oil	180μm
	HCX Plan Apo (PH3)	40/1.25	Oil	100μm
	HCX Plan Apo (PH3)	63x/1.32	Oil	70μm
	HCX Plan Apo (PH3)	63x/1.2-0.7	Water Corr	220μm
	HCX Plan Apo (PH3)	100x/1.4	Oil	90μm
	Dipping Lens	10x/0.3	Water
	Dipping Lens	20x/0.5	Water
	Dipping Lens	40x/0.8	Water
	Dipping Lens	63x/0.9	Water
Software	Leica Confocal Software for normal acquisition, including 3D, co-localisation, multidimensional data/series acquisition (avi movies).
Accessories			Temperature control 37-2 digital heating insert for scanning stage (42mm circular cover slips). Incubator, CTI controller 3700 CO2 IR sensor SPOT RT SE camera, Dell PC with Spot Basic and Advanced software, Image Pro Plus 5.1 image analysis software
Applications	Conventional multi-channel imaging, series acquisition, 3D-6D, live cell imaging, time lapse, ratiometric, FRAP, FRET, reflection, reconstruction.
PC and File Saving	Hewlett Packard, 2G RAM, CD/DVD burner, USB access, Floppy 3.5" 1.44MB

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Laser Capture Microdissection

Location: Research Building CHW, Level 3 SPOT Room 43 4871
Contact: Laurence Cantrill Level 3 Office 9845 3087

The Laser Capture Microdissection facility was opened in July 2003. The microscope was purchased with funding from a Wellcome Grant. The microscope is a PALM MicroBeam system that is mounted on a Zeiss Axiovert 200 inverted microscope. The microscope is equipped with an UV laser that allows regions of interest to be dissected from tissue sections so they can be used in DNA, mRNA or protein analyses. Sections mounted on glass slides can be viewed with bright field or fluorescence. Samples from 1μm - 1mm diameter and of thicknesses up to 70μm can be micro-dissected. Using LPC (Laser Pressure Catapulting) the samples can be collected in the cap of a routine microfuge tube. Sample recovery is easily achieved by simply capping the microfuge tube and centrifuging for a few seconds. Since the system works in non-contact mode and against gravity, contamination by cellular debris is completely eliminated. Furthermore, any unwanted cells or cellular components can be selectively ablated.

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Live Cell Imaging Facility

Location: Research Building CHW, Oncology Research Unit Level 4 Room 44 4875
Contact: Laurence Cantrill Level 3 Office 9845 3087

The Oncology Research Unit's Live Cell Imaging Microscope was purchased in December 2005 with funding from a research infrastructure grant from the NSW Cancer Institute and also by a generous donation from the Oncology Children's Foundation. The system is based on an Olympus IX81 motorised inverted epifluorescence microscope which is fitted with a Märzhäuser motorised stage, a Solent 37°C Incubation Chamber, a Lambda SC Smart Shutter and a Hamamatsu Orca CCD camera. Image capture and processing is controlled via Metamorph software run on a Windows XP workstation with dual high resolution flat screens.

• Bright field
• Fluorescence, HBO 100W Hg short arc lamp, illumination controlled by a fast shutter with open/close speed down to 8ms
• Phase
• DIC

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Digital Imaging and Image Analysis

Location: Research Building CHW, Level 3 SPOT Room 43 4876
Contact: Laurence Cantrill Level 3 Office 9845 3087
Computers: 3 high resolution screens, PCs with 1G ram, iMac (OS9) and iMac (Intel dual processor, OSX)

Scanners

Epson Perfection 1640SU scanner

Printers

Connected to PC's: Epson 890C Photo Stylus Colour Printer.

Connected to iMac: Colour 830U Epson Stylus

Software

• Adobe Photoshop 6.0, CS, CS2 graphic processing
• Adobe PageMaker 6.5
• Adobe Acrobat 5.0 (Reader/Writer)
• Adobe After Effects 5.0
• Adobe Premiere 6.0
• Adobe Illustrator 9.0
• Adobe ImageReady CS, CS2
• Adobe Distiller
• Bioquant Nova Prime image quantification and analysis software
• Image Pro Plus 4.0, 5.1
• Leica IM50 (Image Manager V1.20)
• Leica LCS software for Confocal image analysis
• SPOT Basic/Advanced

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