The Laser Capture Microscopy Core offers a laser capture microscopy service for collecting samples from histological sections or cell cultures.

Director: Christian Waeber, Ph.D.
Phone: (617) 726-0768
Fax: (617) 726-2547
Email: waeber@helix.mgh.harvard.edu

Other Staff:

Research Technician: Igor Bagayev
Phone: (617) 724-0588
Fax: (617) 726-5677
Email: ibagayev@partners.org

Location of Core:

The facility is located at Building 149 13th Street, Room 6028, Charlestown.

Major Equipment:

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 10,000 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.

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-microl, 500-microl, or 1,500-microl sizes). The cap of the
test tube rests on the membrane and has no direct contact with the specimen.

To access the core, please contact Igor Bagayev at (617) 724-0588 or by email at
ibagayev@partners.org.

http://www.massgeneral.org/ncs/neuro_core_lasercapture.htm