How to evaluate a confocal

Hardware
1

Hi All,
What tests do you recommend running when evaluating the performance of a a demo confocal microscope? I’m planning to look at the FWHM of known beads, the evenness of intensity across a field-of-view, and chromatic aberration. Of course I’ll also look at real samples as well. Is there anything else I should test?

Thanks!
Sara

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2

Many of the things you are planning to test are about the quality of the objective and most manufacturer’s have a range of objectives (most of them have some very good ones).

There are many different dimensions across how to evaluate it, image quality is not always the most important one

  • Is this for a facility with many users or a single research group? If it is for a facility, get multiple experienced users to demo it and ask for their feedback ?
  • Live cell imaging: what incubator options do you have with the system? Is the incubator providing consisten temperatures across time and also spatially (no temperature gradients)
  • Automated imaging (if you need it): is it well supported in the software?
  • Does it have a reliable autofocus system ? Does the autofocus system work with the objective and the sample dishes that you are using?
  • Ease and cost of service?
  • Sensitivity and dynamice range of the detector to reduce photobleaching ?

It really depends on your intended application/s. If it is intended for all-round, everyday confocal imaging rather than absolute leading edge resolution it is often the usability (software, controls, etc.) that make the difference between getting good images and bad images for average users in a facility.

3

Those are great things to test for precision, the scopes are so good that they will all test well. I agree with Volker on their points. Testing as many real world samples is important. Try different imaging scenarios, scan methods, push the speed, check sensitivity, try dim and dirty samples, cells, tissues, thick samples, poorly prepped samples. System stability is a big one, image the same sample over a few different days. If timelapsing, focus performance, and repeatability of the stage. How complex is the software? A really BIG one that is often overlooked is managing the image files that some off of the microscope - are they easy to work with across platforms (e.g. Fiji, Imaris, other). It can be a real issue generating 100s of thousands of image files and then having to deal with it. It cannot be stressed enough about the service aspect, many microscopes are great, some may do a few things another doesn’t, and vice-versa - but living with the microscope and dealing with point of contact rep and service will become very important. Upgradability of the microscope, fees to update software, can you purchase additional service contract beyond a certain number of years, etc. Good luck and have fun!

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