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| Format: | Preprint |
| Published: |
2024
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| Online Access: | https://arxiv.org/abs/2406.15621 |
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| _version_ | 1866929395438977024 |
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| author | Chan, Antony C |
| author_facet | Chan, Antony C |
| contents | The 96-eyes instrument is capable of computational extended depth of focus (eDOF) of up to +/- 30 micrometer in the phase channel, and conventional depth of field (DOF) of +/- 5 micrometer in the fluorescence channel. However, it requires minimal plate-to-plate cover glass depth variation to function. Plate depths are measured using a third-party plate scanner (Opera Phenix) grouped by plate types (Greiner UV-Star, Cell-Star, and Eppendorf meniscus-free). The two-dimensional (2D) depth dataset is aggregated through principal component analysis to obtain the top eight dominating 2D surface deformation modes. More than 90% of the variation can be explained by the plate's absolute depth and tilt (Pitch, Gradient-Y, and Gradient-X), followed by (~= 2%) the cover glass's curvature (Curve-Y and Curve-XY). Plate-to-plate average depth and tilt variations are suppressed by a customized kinematic mount anchoring the plate's cover glass at the instrument's imaging plane. The plate's average curvature is compensated by manually aligning all 96-eyes microscope objective lenses to track the plate's surface; an one-off calibration procedure aided by the backlash-free piezo-flexure z-stage. Design validation is conducted in silico, with the proof of concept experiment conducted on the 96-eyes with new mounting bracket retrofits. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_2406_15621 |
| institution | arXiv |
| publishDate | 2024 |
| record_format | arxiv |
| spellingShingle | On the 96-well plate coverglass tilt and curvature suppression in 96-camera imaging system Chan, Antony C Instrumentation and Detectors The 96-eyes instrument is capable of computational extended depth of focus (eDOF) of up to +/- 30 micrometer in the phase channel, and conventional depth of field (DOF) of +/- 5 micrometer in the fluorescence channel. However, it requires minimal plate-to-plate cover glass depth variation to function. Plate depths are measured using a third-party plate scanner (Opera Phenix) grouped by plate types (Greiner UV-Star, Cell-Star, and Eppendorf meniscus-free). The two-dimensional (2D) depth dataset is aggregated through principal component analysis to obtain the top eight dominating 2D surface deformation modes. More than 90% of the variation can be explained by the plate's absolute depth and tilt (Pitch, Gradient-Y, and Gradient-X), followed by (~= 2%) the cover glass's curvature (Curve-Y and Curve-XY). Plate-to-plate average depth and tilt variations are suppressed by a customized kinematic mount anchoring the plate's cover glass at the instrument's imaging plane. The plate's average curvature is compensated by manually aligning all 96-eyes microscope objective lenses to track the plate's surface; an one-off calibration procedure aided by the backlash-free piezo-flexure z-stage. Design validation is conducted in silico, with the proof of concept experiment conducted on the 96-eyes with new mounting bracket retrofits. |
| title | On the 96-well plate coverglass tilt and curvature suppression in 96-camera imaging system |
| topic | Instrumentation and Detectors |
| url | https://arxiv.org/abs/2406.15621 |