PALM DuoFlex Combi System
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.
Samples or cells grown on 35 mm or 50 mm culture dishes and microtitre plates can be imaged with this wide-field live cell imaging platform.
The Navigator tool and scan function allows an overview of the entire (or regions of) plate or dish to be imaged (Movie) , saved as a thumbnail (see image of microtitre plate below) and re-loaded for easy observation and relocation of sample/cell areas in dishes or plates.
Click here to see a movie of how the UV laser cuts and catapults a portion of PEN membrane. A square region of the PEN-membrane on a slide is drawn, cut and catapulted off the slide.
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 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.
The newly installed Zeiss MosaiX module for AxioVision allows automatic scanning of entire samples using motorised stage, followed by pixel-precise image combination of overlapping single images to generate a virtual map using advanced "Stitching" algorithms. It also allows extraction of selected regions and displays as high resolution images for further analysis. Recent upgrades also include a new module for CO2 control to the PALM Combi system. Funding for the purchase of these upgrades was provided by Rebecca L. Cooper Medical Foundation Grant.
The left image shows over 48 (6x8) fields of view taken using a x20 objective and the MosaiX feature of AxioVision software. These images can be automatically stitched together seamlessly allowing a detailed image of larger samples (right image after shading correction and stitching). Hence a single image (of larger samples) can be taken over a larger area that can normally be seen with any objective.
- 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
- Short Guide to Laser Microdissection Using the Palm Combi System