FIGURE 1.4. Examples of microscope system used for in vivo imaging in acute, chronic, and awake preparations.

FIGURE 1.4

Examples of microscope system used for in vivo imaging in acute, chronic, and awake preparations. (left) The imaging setup configured for acute and chronic imaging in anesthetized animals. The system is built around an optics illumination turret of an Olympus BX51WI microscope that is hung from a horizontal X95 optical rail, leaving the underlying area open for the animal and apparatus for stimulus presentation and electrophysiology. The system is shown with the most commonly used 4× macro objective from Olympus (0.28 numerical aperture, 30 mm working distance). Focusing is achieved with a manual focusing ring (LFM-1, Newport). Custom adapters couple the focusing ring to the objective and allow for different objectives to be used while retaining approximate parfocality. The animal preparation, including custom headholder and heating pad, is mounted on translation stages for XY positioning and a vertical linear stage (MVN80, Newport) for rough height adjustment. The stages are robust enough to accommodate a stereotaxic frame and/or micromanipulators for electrophysiology. This entire assembly is mounted on an optical rail and is slid forward of the microscope for the initial surgical procedures. A stereomicroscope can be swung into place for the surgery (not shown). This arrangement minimizes difficulty in arranging the preparation for imaging. For chronic imaging, alignment rods connect the microscope, camera and preparation for precise repositioning of the preparation at different time points (not shown). (right) Configuration used for awake, head-fixed imaging. The microscope and camera setup is identical. A custom head-fixation bar and restraint chamber is positioned below the microscope along with additional apparatus for behavioral training and monitoring. This assembly is mounted on XY translation stages and a vertical jack (Model 281, Newport) for positioning, and is surrounded by a custom enclosure that allows access for the objective. Focusing is achieved manually from outside of the enclosure.

From: Chapter 1, Optical Imaging of Brain Activity In Vivo Using Genetically Encoded Probes

Cover of In Vivo Optical Imaging of Brain Function
In Vivo Optical Imaging of Brain Function. 2nd edition.
Frostig RD, editor.
Boca Raton (FL): CRC Press/Taylor & Francis; 2009.
Copyright © 2009, Taylor & Francis Group, LLC.

NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.