Laser Capture Microscopy
PI: Cristopher Bragg
For reservation, contact the PI or
Phone: 617 - 726-0817
Pager: 617 - 339-6455
or online (http://ppd.partners.org, Pager ID: 22999)
Location: room 6.501A
The SL µCut system consists of an inverted microscope (a Nikon TE-2000S identical to the one used in the Image Analysis Core), a high precision XY-stage, solid state UV-laser (with a wavelength of 350 nm, which guarantees precise cutting without damaging the tissue; it will run maintenance-free for approximately 10000 hours), a fast low-noise digital color camera (for viewing live video and capturing images), computer, and software.
Frozen or paraffin sections are cut (a wide range of tissue thickness can be used) onto a special PET (polyethylene terephthalate) membrane mounted on a 25x75 mm metal frame. Smears can also be prepared on this membrane for cell capture. In addition, cells (including neurons) can be grown on this membrane forming the bottom of special tissue culture dishes to capture live cells for sub-culture.
One of the main advantages of SL µCUT is its stable and easy-to-use software interface. The high precision, motorized XY-stage is controlled through computer mouse, keyboard or stylus/touch screen. An overview of the whole slide is first acquired with the 4x objective and allows easy and fast navigation within the slide. In addition, several positions of the stage can be stored in order to easily return to interesting areas of the section. The laser beam parameters can be adjusted by scrollbars and the parameters are automatically stored and recalled when the system is started again. Other parameters such as focus and camera settings (light intensity) are also stored and automatically recalled when the magnification is changed.
The cutting path of the laser is outlined using either a free hand drawing tool or geometric figure tools such as circles, squares and ellipses (see Figure 1, below). An unlimited number of objects can be outlined, even outside of within the viewing field can be marked and will be automatically cut. It is possible to delete one or several outlines before performing the actual laser capture.
a) Marking several areas for automatic dissection
b) Computer-controlled dissection with UV laser
c) The dissected areas can be easily examined on the collecting cap, for quality control purposes.
Up to four different groups can be defined for cutting. For instance, neurons labeled with a specific antibody can be marked for cutting as group A, and unmarked neurons as group B. The cutting is then done group by group, simply changing the cap in between. The isolated samples are cleanly and efficiently removed using an adhesive film located on the cap of Epperdorf tubes (available in 200-ml, 500-ml, or 1,500-ml sizes). The cap of the test tube rests on the membrane and has no direct contact with the specimen.
More information can be found on the Molecular Machines Inc web site.
Click here to download the SL µCut MMI user manual.
Click here to download a price list for MMI consumables (note that our service contract entitles us to a 5% discount until 2008).
Purchase of this instrument was supported by NIH grant 1S10RR021191-01.