Improved Tracking and Resolution of Bacteria in Holographic Microscopy Using Dye and Fluorescent Protein Labeling
Abstract
Digital holographic microscopy (DHM) is an emerging imaging technique that permits instantaneous capture of a relatively large sample volume. However, large volumes usually come at the expense of lower spatial resolution, and the technique has rarely been used with prokaryotic cells due to their small size and low contrast. In this paper we demonstrate the use of a Mach-Zehnder dual-beam instrument for imaging of labeled and unlabeled bacteria and microalgae. Spatial resolution of 0.3 μm is achieved, providing a sampling of several pixels across a typical prokaryotic cell. Both cellular motility and morphology are readily recorded. The use of dyes provides both amplitude and phase contrast improvement and is of use to identify cells in dense samples.
Additional Information
© 2016 Nadeau, Cho, Kühn and Liewer. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Received: 24 February 2016; Accepted: 31 March 2016; Published: 19 April 2016. This work was supported by the Gordon and Betty Moore Foundation (GBMF) grant numbers 4037 (McGill University) and 4038 (California Institute of Technology). We also thank the Keck Institute for Space Studies for allowing us use of the Tolman/Bacher House, which served as a meeting location for our team on the Caltech campus. We appreciate the contribution of Asphericon, Inc. for fabricating customized optical elements to meet the packaging needs. This work was partially carried out by the Jet Propulsion Laboratory, California Institute of Technology.Attached Files
Published - fchem-04-00017.pdf
Supplemental Material - video_1.avi
Supplemental Material - video_10.avi
Supplemental Material - video_2.avi
Supplemental Material - video_3.avi
Supplemental Material - video_4.avi
Supplemental Material - video_5.avi
Supplemental Material - video_6.avi
Supplemental Material - video_7.avi
Supplemental Material - video_8.avi
Supplemental Material - video_9.avi
Files
Name | Size | Download all |
---|---|---|
md5:63309ba3c7bd00acaa95483721bb593e
|
6.6 MB | Download |
md5:2fa92e1ef535893c82a40417f89e6aa7
|
8.8 MB | Download |
md5:19d886577be81add1aa32799568faeea
|
9.5 MB | Download |
md5:4b331df5d70581ba8553166b97ad0943
|
5.4 MB | Download |
md5:d3bdcd1c3d2b2016f6fbc91b31db72e9
|
1.9 MB | Download |
md5:928e1ab2d4cb69e163637765aa56cb1d
|
4.2 MB | Download |
md5:3d78bccec6e1b9e2440531e720586db9
|
2.1 MB | Download |
md5:4ca2ad5f61ebe4ec6ec53eb00ebb284e
|
6.4 MB | Preview Download |
md5:7bab2eab3abae77bca236b19a57d4822
|
4.8 MB | Download |
md5:3d8ff08245b7301cdbb1bda1a09589d8
|
894.4 kB | Download |
md5:9ff37a63e9aa841e5ce814c78309c920
|
1.2 MB | Download |
Additional details
- PMCID
- PMC4874365
- Eprint ID
- 67854
- Resolver ID
- CaltechAUTHORS:20160610-153959848
- 4037
- Gordon and Betty Moore Foundation
- 4038
- Gordon and Betty Moore Foundation
- Created
-
2016-06-10Created from EPrint's datestamp field
- Updated
-
2021-11-11Created from EPrint's last_modified field