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| Microdissection Applications |
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Laser microdissection from Carl Zeiss provides an intelligent tool in molecular analysis at DNA, RNA and protein levels, as it highly improves sampling of cell-specific tissue.
An innovation in the field of laser microdissection is the isolation of live cells from culture. Individual or small groups of cultured cells, even from primary cultures or stem cell preparations, can be used for direct molecular analysis or recultivation.
Single or few cell analysis on Microarrays: Development of molecular markers for personalized medicine
Malignant cells or other cells of interest might represent a minority in a preparation, so that their gene expres-sion signatures might be hidden by investigating this material. Isolation of a certain single cell or cell population mostly is a prerequisite for a successful description of a cell-type specific gene expression. By Microarray-based gene expression analysis of low cell numbers, even by starting with a minimum amount of only one cell, allows for building the basis for reliable and successful development of a molecular marker. Final goal is a tailored clinical management of the individual patient or groups of patients.
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Proteom analysis: Biomarker discovery and identification in human tumors
The analysis of the proteome is one of the most challenging tasks for the next 20 years. The complexity of the proteome is extremely high, as every organ or even part of it can differ considerably in its protein composition. Performing high quality proteomic studies therefore requires the separation of these functional different tissue areas before analysis. The separation of functional tissue areas can be easily achieved by laser microdissection. Laser microdissected sections from different kind of tumors have been analyzed on ProteinChip arrays. Results have been compared with adjacent “non-tumor” samples. Two differentially expressed proteins were isolated. This technical approach opens up the possibility to find, identify and characterize tumor relevant biomarkers. This way proteins could be identified that are responsible for invasion and metastasis.
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Plant Research: Application of LCM to characterize molecular markers in plant genome analysis and breeding
During the early period of research, classical strategies including comparative anatomy, physiology and embry-ology were employed in genetic analysis to determine inter- and intra-species variability. In the past decade, however, molecular markers characterized by LCM have very rapidly complemented the classical strategies. These markers once mapped enable dissection of the complex traits into component genetic units more precisely thus providing breeders with new tools to manage these complex units more efficiently in a breeding pro-gramme.
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Forensics
DNA typing of biological material has become one of the most powerful tools for suspect identification in forensic medicine and in criminal investigations. STR analysis of DNA is highly discriminating. A complete STR profile of an individual is virtually unique, but even an incomplete profile is powerfully informative.
The use of laser microdissection technology in forensic applications is gaining more and more acceptance around the world. In forensic medicine contact-free microdissection and cell collection without any further manipulation is the ideal approach, particularly to avoid sample contamination and top enrich available material. PALM MicroBeam is the ideal platform to perform that task.
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Chromosomes
Molecular Cytogenetics encompasses all aspects of biology of human, animal, plant and bacterial genomes.
Microscopically visible chromosome rearrangements cannot be determined in any case by conventional cytoge-netic techniques. Complex chromosome rearrangements, small ring chromosomes, marker chromosomes as well as unidentifiable de novo unbalanced translocations for example often need to be investigated on a molecu-lar level. Laser Microdissection of chromosomes is a powerful tool to proceed rapidly from cytogenetic observa-tion to molecular analysis.
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Stem Cell Applications
Stem cells express a variety of specific pluripotency markers, whose expres- sion levels are significant for an undifferentiated state. Manipulation of embryo- nic or adult stem cells may trigger differentiation accompanied by changes in morphology and pluripotency marker expression.
By employing PALM MicroBeam with its unique laser microdissection technology, stem cells can be isolated and propagated without changing their marker expression. Furthermore, stem cell cultures with diversely differentiated cells can be sepa- rated into homogeneous subpopulations, and even embryoid bodies can be sorted.
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Individual Cell Analysis
The structural and functional characteristics of cells are dependent on their specific gene expression profile. The ability to study and compare gene ex- pression at the cellular level will provide valuable insights into the physiology of the cell and is useful in a wide range of research and clinical applications. Even pure cell populations can be far from homogeneous. New methods for single cell analysis have improved the possibilities for detailed analysis of processes inside cells. LMPC combined with different analysis methods allows versatile investigations of individual cells.
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Immunostaining of Living Cells
Many relevant cells in research are commonly complexed with different types of surrounding cells or are rather difficult to distinguish in a mixed culture. The discrimination between those cells often is performed by immunostaining with specific surface markers like fluorescent-labeled antibodies or GFP transfec- tion. Unlike FACS or MACS methods, laser microdissection from Carl Zeiss allows individual adherent cells to be sorted from a mixture under fluorescent illumination and affords the enrichment through clonal expansion after LMPC. Under microscopic control of the specific marker, the selected cell type becomes accessible for characterization within one single purification step without enzyme treatment i.e. of trypsin or harmful selective reagents. Also time consuming repetitive enrichments can be avoided.
Note: Not all applications authorized in all countries.
For details please contact your local dealer. |
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Overview
Laser Microdissection &
Optical Tweezers
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