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Hi All
I use a DMEM based media with phenol red, for neural cell cultures. Luckily it does not seem to matter in our case (td-tomato and EOS2) and has the advantage that we can monitor pH. In fact, it’s how I realised the gas regulator was not working on the new spinning disc....the media was blue-pink after imaging instead of salmon pink.
Best wishes
Julia
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From: Confocal Microscopy List [[log in to unmask]] on behalf of MODEL, MICHAEL [[log in to unmask]]
Sent: 16 January 2019 00:08
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Subject: 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.
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From: Confocal Microscopy List <[log in to unmask]> on behalf of Kilgore, Jason A. <[log in to unmask]>
Sent: Tuesday, January 15, 2019 3:50 PM
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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
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From: Confocal Microscopy List <[log in to unmask]> On Behalf Of Simon Walker
Sent: Tuesday, January 15, 2019 12:13 PM
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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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> On 15 Jan 2019, at 19:03, Chris O'Connell <[log in to unmask]> wrote:
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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?
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> Chris
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