The Bruker Papilio5D light-sheet microscope delivers high-sensitivity imaging for organoids and 3D cell cultures with a high degree of flexibility, enabling researchers to adapt the system to specific sample types, study conditions, and experimental designs.
The system’s horizontal lens geometry with dual-view detection and a choice of Gaussian or Bessel beam illumination, combined with support for simultaneous multi-sample and multi-condition imaging, make Papilio5D a flexible solution for achieving deep and high-resolution imaging while maintaining sample integrity — even on challenging samples and in complex, multi-sample, and multi-condition experiments.
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Papilio5D is photo-efficient for the most delicate samples, using a horizontal geometry with two views/dual-sided detection and a perpendicular light-sheet. This setup is capable of capturing twice as many photons as single-objective detection setups and enables a more comprehensive 3D view of samples.
Fast dual-view acquisition and fusion deliver high-resolution images. Water-dipping high-NA lenses ensure the light sheet is ultra-thin. Area- and line-mode detection also allow data acquisition specialized for different sample requirements.
Together, these features deliver:
Papilio5D can also reach Nyquist sampling, which improves subsequent image processing and enables state-of-the-art deconvolution. Destriping further enhances image quality.
Papilio5D was designed with maximum flexibility in mind. This includes multi-beam and multi-acquisition modes, as well as multi-sample mounting and multi-condition comparison.
Papilio5D users get to choose the best light beam for their experiment. The choice of Gaussian or Bessel beam illumination, both with a beam width below one micrometer, ensures outstanding image quality and great depth-penetration for a wide variety of objects, including dense organoids or zebrafish larvae and fish.
These beams are complemented by two modes of acquisition: area or line mode.
The Papilio5D has a generous sample holder that can hold up to three dishes with two separate chambers each, enabling a total of six experimental conditions to be tested at once. Sample health and integrity are further maintained with environmental control that supports long-term imaging (hours to days). Papilio5D provides control over temperature (20 to 37°C), gas concentration (CO₂ and O₂), and humidity.
| Illumination Modes | Gaussian Beam | Bessel Beam |
|---|---|---|
| Beam Waist Thickness [µm] | 1 bis 6 | 1 (main lobe) |
| Beam Length [µm] | 20 | 350 |
| Pivot Scan | ✓ | ✓ |
| Confocal Line Mode | ✓ | ✓ |
| Acquisition Options | - Gaussian beam - Gaussian with line mode - Gaussian with line mode and pivot scan | - Bessel beam - Bessel beam with line mode - Bessel beam with line mode and pivot scan |
| Illumination Optics | 20x 0.6NA | |
| Detection Optics | Dual 25x 1.1NA / 16x 0.8NA | |
| Multi-Sample/Multi-Condition Experiments | 6 seperate compartments | |
| Magnification Changer | 4 seperate steps | |
| Data processing | Content based dual view registration & fusion | |
| Photomanipulation | CW or pulsed, VIS to NIR | |
Papilio5D supports a wide range of biological imaging applications, from deep imaging of complex 3D organoids to long-term in vivo studies. Its dual-view detection and multi-sample mounting enable comparative experiments, such as tracking regenerative processes in zebrafish under different treatment conditions.
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Time-lapse imaging of zebrafish distal tubules after localized photodamaging can provide novel insights into understanding wound healing processes over time. The design of Papilio5D sample mounting within separate chambers enables imaging under different treatment conditions to be studied at the same time. This allows comparative insights into how pharmaceutical components might impact wound healing.
In this example, regeneration of the distal tubule is studied in the embryonic zebrafish kidney using the cdh17:egfp transgenic line after photodamaging with a photomanipulation module.
| Dual Sided Detection with 2x Objectives | Tube Lens | Magnification | Pixel Size [nm] Orca Flash | FOV [µm] | Pixel Size [nm] Orca Fire | FOV [µm] Orca Fire |
|---|---|---|---|---|---|---|
| Detection 2x Nikon CFI APO LWD 25x 1.10 NA | 100 | 12.5x | 520 | 1065 | 370 | 876 |
| 200 | 25x | 260 | 532 | 184 | 436 | |
| 300 | 37.5x | 173 | 353 | 123 | 290 | |
| 400 | 50x | 130 | 266 | 92* (Nyquist sampling) | 218 | |
| Detection 2x Nikon CFI APO LWD 16x 0.8 NA | 100 | 8x | 813 | 1664 | 575 | 1362 |
| 200 | 16x | 406 | 832 | 288 | 681 | |
| 300 | 24x | 271 | 555 | 192 | 454 | |
| 400 | 32x | 203 | 416 | 144 | 340 | |
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