Re: Live cells in oxygen-scavenging buffer **Vendor response**

[Kilgore, Jason A.]

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**Vendor response**

Hi, Zdenek,

As we say in the ProLong Live manual, it "utilizes Oxyrase technology and contains enzymes from the plasma membrane of naturally occurring E. coli. These enzymes are known for their ability to reduce photobleaching and photodamage in live-cell fluorescent imaging applications."  But we can't say any further about its composition, or how it differs from OxyFluor, beyond saying that we further optimize their technology to meet our standards.

I think it is reasonable to believe the uptake of ProLong Live will affect ROS in the cells in the short term, though I don't think we've confirmed this (such as combining with a ROS indicator like a CellROX dye or H2DCFDA).  And any processes that are dependent upon the cellular ROS state could potentially be affected.

As for how "happy" the cells will be, we've performed testing for viability and cell proliferation with a variety of assays and found no significant effect from ProLong Live even with 48 hours of continuous exposure.  If anyone reading this would like that data, please email me offline and I'll be happy to send it your way.

We haven't tried RI matching, though we have performed imaging at high magnification without any apparent issues.

Hopefully that helps. Cheers,

Jason

Jason A. Kilgore
Technical Application Scientist
Molecular Probes / EVOS Tech Support
Life Sciences Solutions
 
Thermo Fisher Scientific
29851 Willow Creek Rd.
Eugene, OR  97402-9132
1-800-955-6288 then option 4, then option 3, then option 2.
Or dial direct at [phone]
[hidden email]
www.thermofisher.com 

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Dear Listers,
when going through the product sheets of Prolong Live (an anti- photobleaching agent for live-cell fluorescence imaging) and OxyFluor (an oxygen-consuming membrane fraction of E. coli), I couldn't help but notice some similarity...

https://assets.thermofisher.com/TFS-Assets/LSG/manuals/ProLong_Live_Antifade
_Reagent_UG.pdf


https://urldefense.proofpoint.com/v2/url?u=https-3A__www.oxyrase.com_upload_product-5Finserts_lab.0093.v006-5F-2D-5Foxyfluor-5Fpi&d=DwIFaQ&c=q6k2DsTcEGCcCb_WtVSz6hhIl8hvYssy7sH8ZwfbbKU&r=MVp-2yJ1A-yQtCbZ-DE9xd0W6E7srQQpV-yioYjTLyQ&m=eO7GVz7SWDRkFE75lOEIFXLw8hn19BGbHNAsj1ZseFA&s=Ml3Wd3vRT-fwbKsPgzjIT68rYC26h7L6mLaB94jLggg&e=.
pdf
Can someone confirm that Prolong Live is indeed based on OxyFluor?

Which brings up the question: How happy are the cells gonna be in an oxygen- scavenging environment?

Has anyone tried combining this with refractive index matching, such as iodixanol (OptiPrep) or Histodenz or other?

Thanks for any tips or comments!

No commercial interest.


Best, zdenek

--
Zdenek Svindrych, Ph.D.
Research Associate - Imaging Specialist
Department of Biochemistry and Cell Biology Geisel School of Medicine at Dartmouth
email: [hidden email]