*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Mike,

You may also consider doing Raman (or CARS/SRS) if you have access. Lipids give you quite characteristic Raman peaks.

Greetings			Gabor

-----Original Message-----
From: Confocal Microscopy List <[log in to unmask]> On Behalf Of Model, Michael
Sent: 25 March 2022 14:46
To: [log in to unmask]
Subject: Re: EXT: Re: [External] lipid stains

*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
Post images on http://www.imgur.com and include the link in your posting.
*****

Phil -

that was indeed on the plans. But if we could see a membrane by fluorescent staining that might save us the work of doing TEM

Mike

-----Original Message-----
From: Confocal Microscopy List <[log in to unmask]> On Behalf Of Oshel, Philip Eugene
Sent: Friday, March 25, 2022 8:16 AM
To: [log in to unmask]
Subject: EXT: Re: [External] lipid stains

*****
To join or leave the confocal microscopy listserv or to change your email address, go to:
https://nam11.safelinks.protection.outlook.com/?url=https%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FSUBED1%3Dconfocalmicroscopy%26A%3D1&amp;data=04%7C01%7Cmmodel%40KENT.EDU%7C3eba6ea26b8b4dc32b1108da0e594f1e%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637838073996155455%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&amp;sdata=EVUx2PCuc5DRjinfRxyno14yRgxzZmrK3Tfall%2F1cS8%3D&amp;reserved=0
Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cmmodel%40KENT.EDU%7C3eba6ea26b8b4dc32b1108da0e594f1e%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637838073996155455%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&amp;sdata=VazuBzYHTNLvGu7Vjuf8sKP8iB3oofR9iX%2BiFqCrl7c%3D&amp;reserved=0 and include the link in your posting.
*****

Mike,

The membrane may be too thin to stain strongly enough with DiO or DiI to be seen. You could try TEM methods for membranes. Osmium tetroxide preferentially (+/- preferentially) stains membranes, as it likes the C=C double bound, especially conjugated bonds. This can be enhanced by using OsO4 with monomeric tannic acid or ferrocyanide.
Given your question, you'll likely have to end up using TEM methods anyway, to prove these are membrane-bound organelles.

Phil
-------------
Philip Oshel
Imaging Facility Director
Biology Department
1304 Biosciences
1455 Calumet Ct.
Central Michigan University
Mt. Pleasant, MI 48859
989 774-3576 office

-----Original Message-----
From: Confocal Microscopy List <[log in to unmask]> on behalf of "Model, Michael" <[log in to unmask]>
Reply-To: Confocal Microscopy List <[log in to unmask]>
Date: Thursday, March 24, 2022 at 21:05
To: "[log in to unmask]" <[log in to unmask]>
Subject: [External] lipid stains

    *****
    To join or leave the confocal microscopy listserv or to change your email address, go to:
    https://nam11.safelinks.protection.outlook.com/?url=https%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FSUBED1%3Dconfocalmicroscopy%26A%3D1&amp;data=04%7C01%7Cmmodel%40KENT.EDU%7C3eba6ea26b8b4dc32b1108da0e594f1e%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637838073996155455%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&amp;sdata=EVUx2PCuc5DRjinfRxyno14yRgxzZmrK3Tfall%2F1cS8%3D&amp;reserved=0
    Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&amp;data=04%7C01%7Cmmodel%40KENT.EDU%7C3eba6ea26b8b4dc32b1108da0e594f1e%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637838073996155455%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&amp;sdata=VazuBzYHTNLvGu7Vjuf8sKP8iB3oofR9iX%2BiFqCrl7c%3D&amp;reserved=0 and include the link in your posting.
    *****

    Dear List:

    We have found some strange organelle in cultured cells subjected to stress and trying to determine whether it is surrounded by a lipid membrane. If such nonspecific (are they?) lipid stains as DiI or DiO don't stain it, does it make sense to try other lipid stains?
    Thank you!

    Mike

CAUTION: EXTERNAL SENDER Do not click any links, open any attachments, or REPLY to the message unless you trust the sender and know the content is safe.