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Re: HBSS Autofluoresence? vendor reply

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Kilgore, Jason A.

Re: HBSS Autofluoresence? vendor reply

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** Vendor reply **

We've seen the autofluorescence from riboflavin as well. It's part of what prompted us at Thermo Fisher to release our FluoroBrite DMEM and our Live Cell Imaging Solution (a HEPES derivative), which lack the autofluorescent components as well as phenol red (which can partially quench visible wavelength dyes).

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Life Sciences Solutions

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Simon Walker
Sent: Tuesday, January 15, 2019 12:13 PM
To: [hidden email]
Subject: Re: HBSS Autofluoresence?

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Hi. We’ve seen this a lot and come to the conclusion it’s riboflavin. I think it sticks to the glass/plastic on which the cells are growing and briefly fluoresces before bleaching. It’s replenished from the solution, so once the imaging has stopped the signal recovers. Using low riboflavin medium should resolve the issue.
Simon

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> We are trying to image live cultured cells in Hanks Balanced Salt solution without phenol red. It's widefield imaging with a standard Chroma GFP filter set and I'm seeing something I've never observed before. There's very high background all over the field which quickly fades within a few seconds. If I stop the acquisition briefly and resume, the background goes right back up by about 25%. I've never seen this before. What could be the source of this in HBSS, or is it something else?
>
> Chris

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mmodel

Re: HBSS Autofluoresence? vendor reply

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If they are using HBSS, there shouldn't be any riboflavin. I would try HBSS from another bottle.

________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Kilgore, Jason A. <[hidden email]>
Sent: Tuesday, January 15, 2019 3:50 PM
To: [hidden email]
Subject: Re: HBSS Autofluoresence? **vendor reply**

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** Vendor reply **

We've seen the autofluorescence from riboflavin as well. It's part of what prompted us at Thermo Fisher to release our FluoroBrite DMEM and our Live Cell Imaging Solution (a HEPES derivative), which lack the autofluorescent components as well as phenol red (which can partially quench visible wavelength dyes).

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Life Sciences Solutions

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Simon Walker
Sent: Tuesday, January 15, 2019 12:13 PM
To: [hidden email]
Subject: Re: HBSS Autofluoresence?

CAUTION: This email originated from outside of the organization. Do not click links or open attachments unless you recognize the sender and know the content is safe.

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Hi. We’ve seen this a lot and come to the conclusion it’s riboflavin. I think it sticks to the glass/plastic on which the cells are growing and briefly fluoresces before bleaching. It’s replenished from the solution, so once the imaging has stopped the signal recovers. Using low riboflavin medium should resolve the issue.
Simon

Sent from my mobile