SSN-2025-063-JS Super-Resolution Spinning Disc Confocal Microscopy System.pdf

SSN-2025-063-JS Super-Resolution Spinning Disk Confocal Microscopy System 1

Notice of Proposed Sole Source SSN-2025-063-JS

Super-Resolution Spinning Disk Confocal Microscopy System

Colorado State University (CSU) hereby gives notice of its intent to purchase a 3i Marianas Super- Resolution Spinning Disk Confocal Microscopy System via sole source. The spinning disk confocal microscopy system will enable CSU to achieve research objectives of multiple research teams within the department of Biochemistry and Molecular Biology that require rapid fluorescence image acquisition from live cells with super resolution applied rapidly on acquired images. CSU will integrate components from our current 3i spining disk microscope put in service in 2006 with upgrades to integrate the new system with these items which are a laser stack system, v3 fiber switcher, and Phasor which will be upgraded to a new orientation on the new Marianas microscope, the compatibility of the new system with these components enriches capabilities of total internal reflection (TIRF) as well as HiLO imaging and photomanipulation with multiple laser lines. The microscope will become part of a CSU open access core research facility which will serve the major users on the grant as well as CSU’s community of researchers.

RESPONSE REQUIRED FROM VENDORS

RESPONSES TO THIS PROPOSED SOLE SOURCE MUST INCLUDE THE

FOLLOWING:

1) A “Point by Point” response to the requirements as outlined below. This would include but is not limited to; a narrative for each specification/feature, documentation to certify and validate vendor experience (PDF’s or other documents) containing vendor information which can be evaluated by CSU personnel and any other pertinent information concerning the vendor’s equipment. Detailed information is indicated.

2) Vendors must structure their response in the exact order of the requirements above. Responses must be formatted as outlined, followed by a “point by point” discussion of each specification/feature.

3) Colorado State University will be the sole judge of equivalence.

4) All questions pertaining to this request must be submitted online through Rocky Mountain E-

Purchasing System solicitation number SSN-2025-063-JS. Jeb Stuart is the sole point of contact, Jeb.Stuart@colostate.edu

5) FAILURE TO RESPOND AS INSTRUCTED ABOVE WILL AUTOMATICALLY

RESULT IN DISQUALIFICATION.

Firms interested in providing these specialized services must read and be familiar with the below requirements before responding. Responses must be submitted via Rocky Mountain E-Purchasing System (RMEPS)/BidNet Direct, no later than 4:00 P.M. MT on Friday, September 26, 2025.

https://www.bidnetdirect.com/colorado/colorado-state-university. For vendor guides on utilizing RMEPS, please visit: Rocky Mountain E-Purchasing Information. The file name(s) must include “SSN-2025-0063-JS Super-Resolution Spinning Disk Confocal Microscopy System.”

SSN-2025-063-JS Super-Resolution Spinning Disk Confocal Microscopy System 2

UNDER NO CIRCUMSTANCES WILL A TELEPHONE, EMAIL, FAX, OR PAPER

RESPONSE BE ACCEPTED.

Business Need The department of Biochemistry and Molecular Biology requires a spinning disk confocal microscopy system with rapid fluorescence image acquisition from live cells with super resolution applied rapidly on acquired images. The system will enable CSU to achieve research objectives of multiple research teams capabilities of total internal reflection (TIRF) as well as HiLO imaging and photomanipulation with multiple laser lines. The microscope will become part of a CSU open access core research facility which will serve CSU’s community of researchers. Current and future investigations require a technically challenging combination of capturing low-intensity signals from single biological molecules at high spatial resolution and rapid speeds. The system must integrate both a very rapid CMOS camera system and highly sensitive EMCCD camera to provide wide-field, TIRF, SDC, and HILO imaging, along with high magnification and super-resolution of intracellular molecular interactions. The spinning disk illumination system must also provide superior brightness with less phototoxicity than laser scanning microscopes, a critical feature for the long-term live-cell experiments which CSU is investigating. The new system will enhance our imaging capabilities for confocal capture of low-intensity specimens and deliver greater resolution and speed for this imaging.

Required Features Colorado State University requires a super-resolution spinning disk confocal microscopy system that has the following required capabilities which are critical to CSU research requirements.

1. Must provide wide-field, total-internal reflection fluorescence (TIRF), spinning disk confocal

(SDC), and highly inclined and laminated optical sheet (HILO) imaging capabilities for versatile applications, along with super-resolution for tracking intracellular molecular interactions.

2. Must be capable of advanced photomanipulation to provide precise targeting of regions, lines, and points with a diffraction-limited steerable laser spot.

3. Must allow for holographic photomanipulation within TIRF or SDC experiments and provide increased speed by stimulating multiple regions of interest simultaneously rather than by raster scanning in sequence.

4. CSU researchers must be able to have flexibility to choose between holographic stimulation for speed or raster-based approaches when higher laser power is required and these capabilities must be integrated seamlessly, allowing system cameras to be combined with SDC, TIRF, fluorescence recovery after photobleaching (FRAP), z-stacks, and multi-channel imaging within a single experiment, critical for monitoring single molecules in the context of cytoplasmic or nuclear markers with high fidelity.

5. The SDC system must provide capabilities with a large field of view (up to 16mm x 16mm), super-resolution, and a motorized magnification turret to allow users to select the best camera and modality for each experiment.

6. Must be capable of capturing signals from single biological molecules down to single fluorophores at high spatial resolution and rapid speeds.

7. The spinning disk illumination system must provide superior brightness with less phototoxicity than laser scanning microscopes, a critical feature for long-term live-cell experiments.

SSN-2025-063-JS Super-Resolution Spinning Disk Confocal Microscopy System 3

8. Must have a minimum of 4 laser lines including: 405 nm, 488 nm, 561 nm, and 640 nm.

9. Must provide confocal XY resolution ≤ 120 nm to resolve intracellular structures, such as single-mRNA translation sites and actomyosin fibers.

10. Must have confocal Z resolution ≤300 nm to resolve in the axial direction intracellular structures, such as actomyosin fibers.

11. Must have a confocal acquisition speed ≥ 200 frames per second to track individual proteins as they diffuse in the cytoplasm.

12. Must allow for rapid confocal 4D acquisition with 13 z-planes, 3 laser lines, ≤ 1.5s per series to track single mRNA with minimal photobleaching and phototoxicity for live cell imaging.

13. Must have an autofocus system for drift compensation during live imaging.

14. Must have temperature, humidity and CO2 control.

15. Must provide multi-channel photomanipulation for FRAP/photoactivation of any user defined areas in different acquisition modes (TIRF/HILO/Confocal) to monitor the dynamics of transcription factors and single-mRNA translations sites.

16. The system must include training and maintenance on all components to avoid downtime if problems arise.

Market Analysis CSU performed market research and evaluated multiple competing platforms, including Evident, IXplore with Abbelight super-resolution, Zeiss Airyscan systems, and other spinning-disk confocal systems offered by Nikon and Leica. While these alternatives provide useful features, none fully meet the required specifications. The Evident IXplore with Abbelight super-resolution achieves the highest resolution for single-molecule imaging, but it does so only on fixed samples. Multiple CSU users require live-cell imaging, so this system does not meet the required features. Direct comparison of single mRNA molecules on a 3i Marianas system versus a Zeiss Airyscan showed that the 3i system produces approximately three times higher signal brightness. This translates into reduced laser power requirements and significantly less phototoxicity, which is critical for long-term live-cell experiments. Despite their high spatial resolution, the Airyscan instrument CSU already owns could not capture the low-intensity single-molecule samples with the same acquisition speed as the Marianas. The Nikon, Leica, and other vendors using Yokogawa SoRA spinning-disk units cannot provide the dual photomanipulation laser capability required.

Summary The 3i Marianas Super-Resolution Spinning Disk Confocal Microscopy System will enable CSU to achieve research objectives of multiple research teams within the CSU Scientific and Research community that require the features referenced in this sole source notice.