Internet address(es):

Main address: www.au.dk

Purchase of a Super-resolution TIRF microscope

Aarhus University wants to purchase a Super-resolution TIRF microscope.

The laboratory of Jørgen Kjems under the newly-founded Centre for Cellular Signal Patterns (CellPAT), is doing research on how the spatial configuration of cellular receptors influence the how cells recognize and interact with their environment. An important instrument for characterizing the spatial organization of receptors is a super-resolution TIRF microscope. The microscope is primarily used to analyze paraformaldehyde-fixed cells but will also be used to characterize DNA nanostructures in vitro, and for imaging live cells, tracking receptors and ligands.

The TIRF microscope must be capable of doing the following types of characterization:

— super-resolved TIRF imaging to less than 20 nm (xy) resolution,

— fast diffraction-limited single-molecule live-cell imaging of common fluorophores, including particle-tracking,

— fast single-molecule FRET imaging.

TIRF Microscope requirements:

1. 4 high-powered laser lines suitable for STED/dSTORM/PALM SRLM imaging:

— 640 nm (≥ 800 mW),

— 561 nm (≥ 500 mW),

— 488 nm (≥ 300 mW),

— 405 nm (≥ 120 mW).

2. The microscope must, in addition to TIRF illumination, be capable of epifluorescence, HILO, confocal, and SIM/mSIM illumination/imaging modes.

3. The microscope must have less than 40 nm/hour cumulative drift during acquisition. The microscope must be able to detect excessive external vibrations during imaging acquisition.

4. Acquisition software must provide built-in real-time localization analysis and image reconstruction and visualization. This is required for early evaluation of data quality to determine if the sample and imaging parameters are optimal before completing the full, multi-hour acquisition.

5. Acquisition software must provide functions for single-molecule FRET analysis, including tracing of individual FRET molecule and calculation of FRET efficiency and stoichiometry.

6. The acquisition software must feature the ability to program the acquisition using the Python scripting language and import external python libraries. We have in-house experts with years of Python experience, as well as internal Python libraries spanning thousands of lines of code. Translating our expertise and in-house libraries to a different scripting language is unfeasible.

7. The microscope must come with the optical filters needed to image DAPI, EGFP, Cy3B, and Cy5 fluorophores.

8. The microscope, including the TIRF objective, must be temperature-controlled (20–37 ºC), and must be able to switch from 20 º C to 37 ºC in less than one hour.

9. The microscope must provide CO2/Air inlets to control the atmospheric environment during live-cell imaging.

10. The microscope must feature a 100 x 1,4 NA oil immersion apochromatic TIRF objective.

11. The microscope must feature 2 channel simultaneous imaging, with a dichroic filter between the 561 nm and 640 nm laser.

12. The microscope must feature a fast and sensitive camera, with quantum efficiency of at least 80 %, a frame rate of 80 Hz at 1200 × 1200 pixels.

13. The microscope must have built-in auto-focus with the ability to keep the sample plane in focus for at least 6 hours of acquisition.

14. The microscope must be able to resolve a grid of 12 DNA-PAINT docking sites separated by 20 nm on a DNA origami structure using Cy3B as imaging fluorophore.

15. The microscope must be able to save the imaging data in a format compatible with the Open Microscopy Environment standard.

16. The microscope should have a small form-factor, preferably less than 40 cm on each side, since our available lab space is limited.

17. The microscope must receive free software updates for the life time of the microscope.

Market investigation: We have investigated the market by serching the internet and coomunicating with vendors. after the market investigation we are of that conviction that there is only one supplier for this product, especially when we need the above listet.

The laboratory of Jørgen Kjems under the newly-founded Centre for Cellular Signal Patterns (CellPAT), is doing research on how the spatial configuration of cellular receptors influence the how cells recognize and interact with their environment. An important instrument for characterizing the spatial organization of receptors is a super-resolution TIRF microscope. The microscope is primarily used to analyze paraformaldehyde-fixed cells but will also be used to characterize DNA nanostructures in vitro, and for imaging live cells, tracking receptors and ligands.

The TIRF microscope must be capable of doing the following types of characterization:

— Super-resolved TIRF imaging to less than 20 nm (xy) resolution,

— Fast diffraction-limited single-molecule live-cell imaging of common fluorophores, including particle-tracking,

— Fast single-molecule FRET imaging.

TIRF Microscope requirements:

1. 4 high-powered laser lines suitable for STED/dSTORM/PALM SRLM imaging:

— 640 nm (≥ 800 mW),

— 561 nm (≥ 500 mW),

— 488 nm (≥ 300 mW),

— 405 nm (≥ 120 mW).

2. The microscope must, in addition to TIRF illumination, be capable of epifluorescence, HILO, confocal, and SIM/mSIM illumination/imaging modes.

3. The microscope must have less than 40 nm/hour cumulative drift during acquisition. The microscope must be able to detect excessive external vibrations during imaging acquisition.

4. Acquisition software must provide built-in real-time localization analysis and image reconstruction and visualization. This is required for early evaluation of data quality to determine if the sample and imaging parameters are optimal before completing the full, multi-hour acquisition.

5. Acquisition software must provide functions for single-molecule FRET analysis, including tracing of individual FRET molecule and calculation of FRET efficiency and stoichiometry.

6. The acquisition software must feature the ability to program the acquisition using the Python scripting language and import external python libraries. We have in-house experts with years of Python experience, as well as internal Python libraries spanning thousands of lines of code. Translating our expertise and in-house libraries to a different scripting language is unfeasible.

7. The microscope must come with the optical filters needed to image DAPI, EGFP, Cy3B, and Cy5 fluorophores.

8. The microscope, including the TIRF objective, must be temperature-controlled (20–37 ºC), and must be able to switc

Internet address: https://erhvervsstyrelsen.dk/klagenaevnet-for-udbud