Journal article
Light-microscope specimen holder with 3-axis rotation and small-angle control
Journal of neuroscience methods, Vol.221, pp.15-21
01/15/2014
DOI: 10.1016/j.jneumeth.2013.08.026
PMID: 24025262
Abstract
•Specimen holders were developed for light-microscope observation.•They allow multiple-axis rotation with small-angle control.•Specimen tilting and rotation can be corrected while viewing.•They are easily manufactured and installed on a conventional microscope stage.
Although recent developments in methodologies for light microscopy have enabled imaging of fine biological structures, such imaging is often accompanied by two types of problems. One is a tilting of the specimen with respect to the x–y plane (i.e. rotation around the x- or y-axis) such that the sample is not perpendicular to the optical z-axis, and the other is rotation around the z-axis that precludes optimal orientations for imaging and experimentation. These rotation problems can cause optical aberrations and hamper imaging experiments, even when the angular difference from the ideal position is small.
In order to correct for these practical issues, we have developed a specimen holder with 3-axis (x–y–z) rotation for an inverted light microscope. This allows for full-range rotations of 2–4° for x-, y-axes, ∼24° for z-axis, and a small-angle control of <0.1° for either axis.
Using this device, we observed the cultured hippocampal neurons stained by immunofluorescence for a dendritic marker, or the sub-resolution fluorescent beads plated on a glass coverslip. The rotations and associated problems could be manipulated, while viewing the specimens by laser-scanning confocal microscopy.
This tilting/rotation device is easily manufactured and installed on a conventional microscope stage without requiring changes to the existing optical components. Similar devices with full capability have not been available.
It will be useful for imaging experiments with biomedical applications.
Details
- Title: Subtitle
- Light-microscope specimen holder with 3-axis rotation and small-angle control
- Creators
- Sadahiro Iwabuchi - Department of Molecular Physiology and Biophysics, University of Iowa Carver College of Medicine, Iowa City, IA, USAJin-Young Koh - Department of Molecular Physiology and Biophysics, University of Iowa Carver College of Medicine, Iowa City, IA, USAMichael Wardenburg - Medical Instruments Shop, Engineering Services, University of Iowa Hospitals and Clinics, Iowa City, IA, USAJames D Johnson - Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, United KingdomN. Charles Harata - Department of Molecular Physiology and Biophysics, University of Iowa Carver College of Medicine, Iowa City, IA, USA
- Resource Type
- Journal article
- Publication Details
- Journal of neuroscience methods, Vol.221, pp.15-21
- Publisher
- Elsevier B.V
- DOI
- 10.1016/j.jneumeth.2013.08.026
- PMID
- 24025262
- ISSN
- 0165-0270
- eISSN
- 1872-678X
- Language
- English
- Date published
- 01/15/2014
- Academic Unit
- Molecular Physiology and Biophysics; Otolaryngology
- Record Identifier
- 9984025424802771
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