Microdissection instrumentation

User Guide Veritas Microdissection Instrument. Arcturus Bioscience, Inc Mountain View, CA. 

This chapter summarizes our experience with this approach and describes some of the critical parameters involved in the preparation of rat neural tissue for LCM, with particular emphasis on protocols applied to the Arcturus Autopix LCM. This instrument has been superceded by more recent models, which are now marketed and distributed by MDS Analytical Technologies (http // www.moleculardevices.com). The principles described here, however, are applicable to tissue preparation for most LCM instruments. We describe two basic protocols for the isolation of samples of rat brain, first from unstained microdissected regions, and second, from brain cells that express specific antigens identified by immunostaining. In addition, we compare the effects of different fixation conditions on tissue recovery and RNA content using real-time QPCR. 

Fig. 1. Roadmap image of the entire slide. Automated Laser Capture Microdissection (LCM) instruments display an image of the entire slide that serves as a reference image for determining the areas of tissue to be microdissected. The tissue shown on the roadmap image is coral. LCM can be performed on a variety of tissue types, including plant, coral, shellfish, and mammalian tissue.

Fig. 2. UV cutting laser tools. (A) The UV cutting laser ablates tissue around the perimeter of the desired area. The capture laser is used in conjunction with a Laser Capture Microdissection (LCM) cap to remove the desired tissue section. The LCM cap is bonded, or tacked, to the cut tissue within the perimeter of the cut section, effectively removing the desired cellular area from the tissue section. (B) A free-form annotation tool is also available to select any size or shaped region for ablation/cutting with the UV laser. (C) Circular regions of defined diameter may be ablated/cut with the UV laser-equipped Veritas instrument.

Fig. 5. Live video microdissection. Veritas instruments have the capacity to microdissect directly from the live video image. Various cell populations may be annotated during a single session, matched to a capture group and microdissected on separate Laser Capture Microdissection (LCM) caps. These features are beneficial for morphological differentiation of cells in heterogeneous sections as well as the ability to select multiple cell populations for microdissection from one tissue section.

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