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Active focus stabilization for upright selective plane illumination microscopy.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Hedde, Per Niklas Gratton, Enrico |
| Copyright Year | 2015 |
| Abstract | Due to its sectioning capability, large field of view, and minimal light exposure, selective plane illumination microscopy has become the preferred choice for 3D time lapse imaging. Single cells in a dish can be conveniently imaged using an upright/inverted configuration. However, for measurements on long time scales (hours to days), mechanical drift is a problem; especially for studies of mammalian cells that typically require heating to 37°C which causes a thermal gradient across the instrument. Since the light sheet diverges towards the edges of the field of view, such a drift leads to a decrease in axial resolution over time. Or, even worse, the specimen could move out of the imaging volume. Here, we present a simple, cost-effective way to stabilize the axial position using the microscope camera to track the sample position. Thereby, sample loss is prevented and an optimal axial resolution is maintained by keeping the sample at the position where the light sheet is at its thinnest. We demonstrate the virtue of our approach by measurements of the light sheet thickness and 3D time lapse imaging of a cell monolayer at physiological conditions. |
| Starting Page | F926 |
| Ending Page | F937 |
| Page Count | 1 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://cloudfront.escholarship.org/dist/prd/content/qt1408876j/qt1408876j.pdf |
| PubMed reference number | 26072829v1 |
| Alternate Webpage(s) | https://doi.org/10.1364/OE.23.014707 |
| DOI | 10.1364/oe.23.014707 |
| Journal | Optics express |
| Volume Number | 23 |
| Issue Number | 11 |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | CDISC SEND Biospecimens Terminology Entity Handling - upright Gradient Hearing Loss, High-Frequency Heating Mammals Microscope Device Component Sectioning technique Time-Lapse Imaging |
| Content Type | Text |
| Resource Type | Article |
