Detection
Flamingo microscopes feature fast & low-noise camera, such as the pco.panda 4.2. We integrate several detection lenses, e.g. selected models of the Nikon CFI60 and CFI75 water dipping series. Each lens can be combined with tube lenses of different focal length to adapt magnification to the imaging needs. Emission filters for extracting fluorescence signals are mounted in custom-built motorized filter wheels in the detection path.
FIELD OF VIEW & SPATIAL SAMPLING
Several detection lenses can be used to adjust the field of view and the spatial sampling. Representative images and numbers shown here assume a camera chip with 2048×2048, 6.5 μm pixels. The field of view can be reduced by cropping the camera chip to increase acquisition speed or reduce data size, and several fields of view can be combined to cover a larger area (tiling).
| Detection Lens | Lateral Resolution | 200 mm Tube Lens | 300 mm Tube Lens | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Magnification | Field of View | Pixel Size | Spatial Sampling | Magnification | Field of View | Pixel Size | Spatial Sampling | ||
| Nikon 10x/0.3 | 833 nm | 10x | 1330 µm | 650 nm | 1.3 | 15x | 887 µm | 433 nm | 1.9 |
| Nikon 16x/0.8 | 313 nm | 16x | 831 µm | 406 nm | 0.8 | 32x | 416 µm | 203 nm | 1.5 |
| Nikon 40x/0.8 | 313 nm | 40x | 333 µm | 163 nm | 1.9 | 60x | 222 µm | 108 nm | 2.9 |
SAMPLE MOVEMENT
We facilitate PI L-505 or PI M linear stages for motorized sample translation and enable motorized sample rotation using PI U-651 or U-628 stages. The Flamingo can record z-stacks and supports multi-position recordings and tiled acquisition. In an L-, T- or X-SPIM configuration, the vertically mounted sample can be rotated around its central axis. In a V-SPIM configuration, the surface with the sample resting on top can be rotated and two orthogonal z-stacks can be recorded.
ILLUMINATION
Laser illumination in Flamingo light sheet microscopes is achieved by using fibre-coupled laser modules, such as a Toptica iChrome CLE. Static Gaussian light sheets are formed by cylindrical lenses and feature the options to pivot the sheet around the center of the field of view (mSPIM) and illuminate the sample from two sides in T- and X-SPIM configurations. We use an RGB LED for brightfield illumination.
MICROSCOPE CONTROL
Each Flamingo has an integrated computer controlling the hardware and coordinating imaging experiments. The researcher connects remotely to the Flamingo to manually control the microscope and to set up experiments. A computer running macOS 10.12 or later is required for this step.
DATA STORAGE
Flamingo image data is streamed from the camera to computer memory and, in a parallel thread, saved to external USB data storage. For reliable imaging sessions we recommend SSDs mounted in a USB 3.1 enclosure.
DIMENSIONS
A Flamingo in T-SPIM configuration is about 350 mm in diameter and 500 mm tall. The control box is 480 x 380 x 275 mm in size. Microscope and control box each fit in a Pelican 1660 Protector Case. Additional accessories are kept in a Pelican 1485 Air Case. All cases comfortably fit in a compact car, such as a Subara Crosstrek or a Volkswagen Golf, with the rear seats folded down.
The Flamingo #2 requires a workspace of at least 90 x 70 cm (35 x 28″). We recommend a workspace of at least 130 x 70 cm (50 x 28″) to have additional space for samples and laptop. The L-/T-/X-SPIM needs about 510 mm (20″) of clearance.
