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Exp Cell Res. 2015 Oct 1;337(2):249-56. doi: 10.1016/j.yexcr.2015.07.011. Epub 2015 Jul 17.

A mechanical microcompressor for high resolution imaging of motile specimens.

Author information

1
University of the Sciences Chemistry and Biochemistry, 600 S 43rd Street, Philadelphia, PA 19104, United States.
2
Marine Biological Laboratory, 7 MBL Street, Woods Hole, MA 02543, United States.
3
Texas A&M University, Department of Biology, College Station, TX 77843, United States.
4
University of the Sciences, Department of Biological Sciences, 600 S 43rd Street, Philadelphia, PA 19104, United States. Electronic address: c.janetopoulos@usciences.edu.

Abstract

In order to obtain fine details in 3 dimensions (3D) over time, it is critical for motile biological specimens to be appropriately immobilized. Of the many immobilization options available, the mechanical microcompressor offers many benefits. Our device, previously described, achieves gentle flattening of a cell, allowing us to image finely detailed structures of numerous organelles and physiological processes in living cells. We have imaged protozoa and other small metazoans using differential interference contrast (DIC) microscopy, orientation-independent (OI) DIC, and real-time birefringence imaging using a video-enhanced polychromatic polscope. We also describe an enhancement of our previous design by engineering a new device where the coverslip mount is fashioned onto the top of the base; so the entire apparatus is accessible on top of the stage. The new location allows for easier manipulation of the mount when compressing or releasing a specimen on an inverted microscope. Using this improved design, we imaged immobilized bacteria, yeast, paramecia, and nematode worms and obtained an unprecedented view of cell and specimen details. A variety of microscopic techniques were used to obtain high resolution images of static and dynamic cellular and physiological events.

KEYWORDS:

High-resolution imaging; Immobilization; Microbes; Protozoa

PMID:
26192819
PMCID:
PMC4696872
DOI:
10.1016/j.yexcr.2015.07.011
[Indexed for MEDLINE]
Free PMC Article

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