Atch_5_-_Salient_Characteristics.pdf

Attachment 5 - Line Item 5

Salient Characteristics

1. AIRYSCAN-Capture image from broad light source improves resolution. System must have

Airyscan technology (32 channel GaSP) with spatial detection. Confocal laser scanning microscopes (CLMs) are renowned for their sectioning capability. This feature is enabled by the use of pinhole, which rejects out-of-focus light. Less appreciated, on the other hand, is the gain in lateral resolution by this type of microscopes for one obvious reason. As the pinhole is closed to improve the resolution, less light can reach the detector leading to images with poor signal-to noise rations (SNR). Therefore CLSMs are operated with a pinhole diameter of around 1 Airy unit (AU), sacrificing resolution for SNR. But with a clever detector design one can solve this conundrum. By using a sub-Airy detector element array instead of a single point detector, it is possible to collect light more efficiently boosting the strengths of confocal. The extra photon budget can be diverted towards increasing the sensitivity, the scanning speed or the resolution of the microscope.

2. ELYRA P.1 --The system must contain an electronically integrated 3D-PALM technology that enables super-resolution photo activated localization microscopy (PALM) in 3D. With

Elyra P.1 and 3D-PALM, researchers can capture highly resolved structures in 3D using photo switchable proteins, while treating the sample so gently it stays fit for long-term observation.

Detection with an effective resolution down to 20 nm shows substructures and patterns where conventional light microscopy will simply show co-localization.

3. Scanhead configuration - The microscope system must be configurable all at the same time with 405nm, 458nm, 488nm, 514nm, 56lnm, 59lnm, and 633nm lasers (all the lasers are capable of being used in real time at the same time).

4. Scanner-Microscope must contain two independent, galvanometric scanning mirrors with ultrashort line and frame fly back.

5. Total Scanning Resolution: The microscope must have 4Xl to 8192x8192 pixels AND for multiple channels be continuously adjustable

6. Scanning Speed and Resolution: Microscope must have 19x2 speed levels and be able to capture up to 13 images with 512x512 pixels)

7. Scanning Zoom: microscope must be able to have a scanning zoom of 0.6x40 zoom which can be digitally adjustable in 0.1 increments (for the Zeiss this capability is contained in the

Axioexaminer (0.67x to 40x)

8. Scanning Rotation must be rotated freely 360 degrees and be adjustable in increments of one degree and possess a freely adjustable xy offset

9. Detectors: the microscope must possess 34 spectral detection channels (GaSP and/or PMT)

10. Spectral Detection: 34 simultaneous confocal reflected light channels and be either GaSP or PMT based, freely adjustable spectral detection area down to 3 nm

11. Real-Time Electronics: 1) The microscope, the scanning module, the adjustable accessory control, data acquisition, synchronization management must be able to be accomplished through real-time electronics 2) the microscope must possess oversampling readout logic and

3) data transfer between real-time electronics and user PC via Gigabit Ethernet with the ability to evaluate the data online during image acquisition

12. Pinholes: The microscope must contain a master pinhole with a preset size and position that can be adjusted as desired for multitracking and short wavelengths such as 405nm

13. Beam Path: the beam path of the microscope must contain exchangeable twin gate beam splitter with up to 100 combinations of excitation w a v e l e n g t h s and outstanding laser line suppression. The microscope must contain a manual interface port for external detection modules (such as BiG.2, Airyscan, third party detectors, internal detection, with spectral signal separation and signal recycling loop for compensation of polarization effects

14. Multitracking: The microscope must change excitation lines when recording multiple fluorescence’s for the purpose of minimizing signal crosstalk and increasing dynamic range

15. Z Drive for Depth: The smallest increment of the Z drive must be able to reach <25 nm

16. Optional Software: The microscope must be able to accommodate all the following additional software: 3D VisArt, Deconvolution, ROI-HDR, Physiology, FRET, FRAP efficiency analysis, Visual Macro Editor, RICS Image Correlation, Experiment Designer and

Macroenvironment.