What is Laser Capture Microdissection (LCM)? Three LCM machines are available in UCHC Core. Two of they are ArcturusXT™ Laser Capture Microdissection (LCM) System. The LCM system uses a laser beam to activate a special transfer film. The film bonds to cells that have been identified and targeted by microscopy. From this film, macromolecules can be isolated and studied in downstream analyses. An infrared (IR) laser is used for single cells and small populations of cells, while an ultraviolet (UV) laser can be used for large populations of cells. Starting tissue sections (5-7um) can be frozen or fixed/paraffin embedded, however it is ideal to use frozen sections for most downstream applications. |
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Why use LCM in Gene Expression Study? To reveal accurate, cell-type specific expression profiles that otherwise might be obscured in mixed cell samples. Technology isolates homogeneous populations of cells from tissue sections. Cells can be collected based on morphologic or immunostaining or enzyme staining. Machine can recognize color (stains) or fluorescence Cells are harvested for RNA, DNA or Protein analysis |
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What is different between IR Capture and UV Cut?
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What kind tissue section and slides can be used for LCM? Both of Frozen tissue section (fresh and fixed frozen section) And Paraffin section can be used for LCM. Both of mineralized tissue (bone and tooth) and non-mineralized tissue can be used for LCM. Types of slide for LCM: • No charged pre-cleaned plain glass slide Required dehydrated section and IR capture only. • CryoJane tape system Required dehydrated section and IR only capture. Particularly useful for mineralized tissue. • LMD Flim non-dehydrated section preparations and UV capture only. Particularly useful for mineralized tissue. • PEN membrane glass slide Required dehydrated section and IR and UV capture. • PEN Membrane Frame slide. non-dehydrated section preparations and IR and UV capture. |
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What LCM machines are available to use in LCM Facility? There are 3 LCM microscopies in UCHC LCM Core. The ArcturusXT™ microdissection system (life Technologes, Inc) a unique combination of IR laser-enabled LCM and UV Laser Cutting in one platform. The open, modular design of the ArcturusXT™ system provides unparalleled research flexibility and versatility. The LCM microscope has brightfield (including Phase, DCIM) as well as fluorescent capabilities.
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What LCM services are available in LCM Core? Services available include: 1) Tissue processing (Tissue isolation, tissue embedding, section, staining) 2) Laser capture of specific cell types or cellular structures 3) Extraction of RNA, DNA or protein 4) Consultation for experimental design 5) Assistance with grant preparation. Hourly service charges are based on the complexity and duration of the job. We will train you to use the machine, and provide technical assistance and reagents for dealing with LCM-captured material for subsequent molecular analysis. |
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How to use the Laser Capture Microdissection Service: Schedule an initial appointment with the service director or co-director to discuss your project, including what the ultimate downstream application will be and the general feasibility of your project. Registration for first time users, establish an account by filling out and submitting a Customer Information Form that you can download or fill the form from our FORM page. Sign-up for a three or six–hours training session to learn how to prepare section for LCM and how to use the LCM Machine and (or) how to do RNA evaluation. If you have frozen or paraffin sections of your sample prepared, you can use one of them during your training session. If not, we will provide tissue for you to use for training. Schedule Machine usage for independent user who finished training session. Schedule usage time sign in LCM facility website via the website of UConn ResearchCores. |