Re: Comments welcome on how super-resolution techniques have changed localization conclusions made with confocal microscopy

[Brandi Bickford]

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Hi Madison,

I have used the  confocal microscopes many times for localization. You have to take into account stereology effects, which can become a bear. Essentially you  need to measure the distance between the two signals to prove that they are not just close to one another. I would suggest Visiopharm who collaborates with Olympus to help explain this to you, as I have not done confocal for some time. They have an app available for this particular issue. Digital pathology takes up all of my work effort hours. I just found an absolutely wonderful, unquestionable application for localization that we are putting in place. The 3D Histech, at least the P1000, can create 3D images which will confirm irrevocably weather the signal is in fact colocalized and specifically where. This is amazing. One of our researchers asked me about this in the hall one day, he had heard about it and did not believe this capability existed, he was making a joke. I emailed me discreetly that we have one down the hall that can do this , and do it quickly. I would never rely on confocal now that this leaves no question and no discussion about difficult measurements. Hope this is of help.

Sincerely,
Brandi Bickford
WFUBMC, dept. of Pathology
Digital Pathology/Virtual Imaging
 

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Madison Yemc
Sent: Friday, December 20, 2019 11:27 AM
To: [hidden email]
Subject: Comments welcome on how super-resolution techniques have changed colocalization conclusions made with confocal microscopy



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Hello. My name is Madison Yemc, and I am currently preparing my master’s thesis under the direction of Dr. Bob Price. As part of my thesis, I am conducting a literature review to investigate how super-resolution techniques may have changed our analysis and interpretation of the colocalization of spatially related molecules and the resulting functional implications. Previously, many studies have implied functional interaction based on confocal images of red + green = yellow. Now, super-resolution techniques show these molecules may not be as closely related as previously thought.

Any comments on how super-resolution techniques have changed our previous conclusions made with standard confocal microscopy are welcome.



All responses are very much so appreciated in advance! Thank you for your time.



Kind regards,

Madison Yemc



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Madison Yemc

University of South Carolina School of Medicine

Candidate for MS Biomedical Science  |  Class of 2020

412-715-6945  |  [hidden email]

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