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.
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
What is different between IR Capture and UV Cut?

Type of Dissection IR Capture UV Cut
Type of Laser

IR Laser

  
Slide Required *Plain glass slides *Membrane slide
  *CryoJane slides *Frame slide
  *Membrane slide *LMD film
  *Frame slide  
Mechanism IR Laser transfer cells into a thermoplastic polymer UV Laser cut the membrane or  film with tissue and collected into cap by IR capture
Advantages IR Laser does not damage tissue Fast dissections
  Good for small targets (cells) For big areas of target and clusters cells
    Able to dissect thick section (~20µm)
    Able to dissect large amount of cells
Disadvantages Slow than UV cut Require tissue mounted on special slides or film
    High energy laser could affect biomolecules in very small target dissections
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.

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.


Filters Include

Emission

Excitation

Red

>655 nm

570-630 nm

Green

>565 nm

503-548 nm

Blue

>410 nm

455-495 nm

Yellow

380-420 nm

540-570 nm
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.
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.