Laser Microdissection

• PALM DuoFlex Combi System
• Arctursus LCM

PALM DuoFlex Combi System

Room N135, Anderson Stuart Building

IT’S HERE! The P.A.L.M. DuoFlex Combi System purchased from Carl Zeiss is an integrated laser microdissection, optical tweezers and microscope system for live cells and fixed material. For laser microdissection, it makes use of high precision UV laser cutting together with a unique laser microdissection pressure catapulting (LMPC) technology to further eliminate contaminating contact between the sample and the collection device. The end-result is high-purity specimens that can be processed further for DNA, RNA or protein isolation. The operator can choose from a variety of collection vessels for different types of experiments. Selected areas can be transported into the collection vessel via LMPC with just a click of the mouse! Control of experiments is done efficiently with the PALM RoboMover. The system has both high-end fluorescence and live cell isolation capabilities. The AxioObserver microscope platform makes it an ideal arrangement for live cell work.

This dedicated instrument for contact-free manipulation of microscopic particles also combines high performance imaging functionality with optical trapping. This PALM Combi system has “optical tweezers” that requires near-infrared (NIR) laser manipulation (1 fixed, 1 moving) and allows both non-contact and non-damaging micro-manipulation of small particles including cells as well as the possibility of "optically stretching" cells and force measurements. Thus, living cells can be manipulated either in isolation or amongst other cells without damage to their structure or function.

The new Force Measurement software module enables the quantitative measurement of the forces of microscopic specimens, such as cell elasticity, molecular binding properties or plasma membrane features in conjunction with the PALM MicroTweezers.

Funding for this integrated system was provided by Clive & Vera Ramaciotti Equipment Grant 2009, NHMRC Equipment Grant 2009, Cancer Institute NSW Equipment Grant 2009 and the Bosch Institute.

• AutoLPC from glass slides
• RNA extraction from FFPE sections
• RNA extraction from frozen sections
• RNA handling
• Cell Culture, non-contact Laser Capture Microdissection (LCM) and Recultivation
• Chromosome Preparation
• DNA handling
• FISH Hybridization
• Immunofluorescence on frozen sections

Arcturus Laser Capture Microdissection

Room 663a, Blackburn Building

The Laser Capture Microdissection Facility was opened on 18th August 1999. The Arcturus PixCell IIe, valued at $200,000, was funded by the National Health & Medical Research Council and the University of Sydney major equipment fund. In June 2003, a $100,000 upgrade was facilitated by funds from the University of Sydney Sesquicentenary fund, Rebecca L. Cooper Foundation (equipment) and Clive and Vera Ramaciotti Foundation (equipment).

The advanced microscope is equipped with an infra-red laser that removes regions of interest from tissue sections, which can be used in downstream DNA, mRNA or protein analyses. Sections mounted on glass slides can be viewed with bright field or fluorescence.

Optimal specimen preparation is vital in order to achieve successful laser capture of cells. Some general rules apply:

• Tissue processing must be rapid in order to minimise degradation of macromolecules
• Ideally, sectioning, processing and laser capture should be conducted on the same day
• Material must be dehydrated thoroughy using 100% ethanol (with Molecular Sieve) and xylene (to remove the ethanol)

User charges

• The facility operates on a cost recovery basis and provides preliminary training, please refer to current LCM costs

Illumination sources

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

Objectives

• UPlanFl 4x/ 0.13
• UPlanFl 10x/ 0.30
• LCPlanFl 20x/ 0.40
• LCPlanFl 40x/ 0.60 infinity corrected

Fluorescence filters

• Blue, green and red filters, click here for filter specifications

Camera

• Hitachi Digital Model KP-D590P
• 1/2 inch single chip CCD colour camera

• CD
• Zip disc
• Floppy disc 3.5" 1.44MB