Imaging of small biological specimens and microorganisms that are living and moving is often hampered by a traditional microscope’s small field of view at high resolution. This paper discusses a new optical microscope design, called the Adaptive Scanning Optical Microscope (ASOM), which uses a deformable mirror combined with a custom scanner lens to effectively enlarge the field of view. Using a high speed scanning mirror in a post-objective configuration, the ASOM captures a complete image (not a single point) at each scan position and assembles image mosaics on the fly. Consequently, this microscope offers advantages when compared to moving stage based approaches or confocal microscopes. Whereas previous work on imaging motile organisms has primarily focused on tracking only one temporally challenging specimen at a time within a single field of view, this microscope is well suited for tracking multiple moving organisms or monitoring larger organisms at both the full animal and single cell levels simultaneously. In studies requiring manipulation, probing, or sensing, the ability of the microscope to automatically monitor several regions of the specimen without agitating the workspace is particularly advantageous. Using a low cost prototype of the ASOM, we illustrate the basic capabilities of the instrument by imaging multiple living and freely moving Caenorhabditis elegans nematode worms. In addition to transmitted, reflected, and epifluorescent illumination modes, we have also integrated an LED light source that can be rapidly turned on and off in synchronization with the scanning to minimize unnecessary light exposure to the specimens.
Photonics West, San Jose, Jan, 2007.