Imaging Sourdough batter
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Colleagues, Here's something to chew on. This is not strictly a confocal question, but applies to widefield as well. Some folks here are interested in analysis of the time course of the maturation of sourdough, and have asked us to help them prepare samples of the batter for analysis of, say, yeast content. Have any of you had any experience with this sort of material? We have thought about making frozen sections of the batter at different times after initiation, and staining with DAPI. I'm curious about fixation (note that there are some interesting pH issues here), and quantitation methods. Any suggestions will be appreciated. Joel Joel B. Sheffield, Ph.D Department of Biology Temple University Philadelphia, PA 19122 Voice: 215 204 8839 e-mail: [hidden email] URL: *https://bio.cst.temple.edu/~jbs/ <https://bio.cst.temple.edu/%7Ejbs/> <http://tinyurl.com/khbouft>* |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** GFP expressing yeast. Happy belated St Patrick's Day. Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY 10016 C: 914-309-3270 [hidden email] http://nyulmc.org/micros http://microscopynotes.com/ -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Joel Sheffield Sent: Tuesday, March 20, 2018 3:48 PM To: [hidden email] Subject: Imaging Sourdough batter Colleagues, Here's something to chew on. This is not strictly a confocal question, but applies to widefield as well. Some folks here are interested in analysis of the time course of the maturation of sourdough, and have asked us to help them prepare samples of the batter for analysis of, say, yeast content. Have any of you had any experience with this sort of material? We have thought about making frozen sections of the batter at different times after initiation, and staining with DAPI. I'm curious about fixation (note that there are some interesting pH issues here), and quantitation methods. Any suggestions will be appreciated. Joel Joel B. Sheffield, Ph.D Department of Biology Temple University Philadelphia, PA 19122 Voice: 215 204 8839 e-mail: [hidden email] URL: *https://urldefense.proofpoint.com/v2/url?u=https-3A__bio.cst.temple.edu_-7Ejbs_&d=DwIBaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=MW_N4CU7UnKTXChJnhnUeuHJsCyqA4kASBs7wnkfg1s&s=2fg-ZR-llePOTXxdTNf33ptgPzQa9xUTu7mLY1HqfBg&e= <https://urldefense.proofpoint.com/v2/url?u=https-3A__bio.cst.temple.edu_-257Ejbs_&d=DwIBaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=MW_N4CU7UnKTXChJnhnUeuHJsCyqA4kASBs7wnkfg1s&s=uS4AtkP7JjxSEcqxPqjAEwztKUFkFG29h2326nOw_Wg&e=> <https://urldefense.proofpoint.com/v2/url?u=http-3A__tinyurl.com_khbouft&d=DwIBaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=MW_N4CU7UnKTXChJnhnUeuHJsCyqA4kASBs7wnkfg1s&s=HoPUZltmFsMe-SkpzCK_xNzPn1VIKc_bWpSPXmu3q08&e=>* ------------------------------------------------------------ This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email. ================================= |
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In reply to this post by JOEL B. SHEFFIELD
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Joel, No experience with sourdough except as a consumer, but it sounds like a cool project. If you want to preserve air pockets, then frozen sections make sense. To simply measure micro-organism content, could you just smear some starter/batter on a slide, as in a Pap or blood smear? Possibly, acridine orange would work to stain the bacterial DNA -- in this paper (found via Google), blood smears were stained after a simple MeOH fixation. http://jcm.asm.org/content/11/3/281.full.pdf Calcofluor should light up the fungal cell walls (specific for chitin; use the DAPI channel). Hope this helps, Theresa ------------------------------------ Theresa Swayne, Ph.D. Associate Research Scientist Manager, Confocal and Specialized Microscopy Shared Resource<http://hiccc.columbia.edu/research/sharedresources/confocal> Herbert Irving Comprehensive Cancer Center Columbia University Medical Center 1130 St. Nicholas Ave., Room 222A New York, NY 10032 Phone: 212-851-4613 [hidden email]<mailto:[hidden email]> From: Confocal Microscopy List <[hidden email]> on behalf of Joel Sheffield <[hidden email]> Reply-To: Confocal Microscopy List <[hidden email]> Date: Tuesday, March 20, 2018 at 3:51 PM To: "[hidden email]" <[hidden email]> Subject: Imaging Sourdough batter ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Colleagues, Here's something to chew on. This is not strictly a confocal question, but applies to widefield as well. Some folks here are interested in analysis of the time course of the maturation of sourdough, and have asked us to help them prepare samples of the batter for analysis of, say, yeast content. Have any of you had any experience with this sort of material? We have thought about making frozen sections of the batter at different times after initiation, and staining with DAPI. I'm curious about fixation (note that there are some interesting pH issues here), and quantitation methods. Any suggestions will be appreciated. Joel Joel B. Sheffield, Ph.D Department of Biology Temple University Philadelphia, PA 19122 Voice: 215 204 8839 e-mail: [hidden email]<mailto:[hidden email]> URL: *https://bio.cst.temple.edu/~jbs/ <https://bio.cst.temple.edu/%7Ejbs/> <http://tinyurl.com/khbouft>*<http://tinyurl.com/khbouft%3e*> |
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In reply to this post by JOEL B. SHEFFIELD
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Joel, We've had one person imaging dough who was using Nile Red and Acridine Orange. It worked well. Cheers, Jacqui -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Joel Sheffield Sent: Wednesday, 21 March 2018 8:48 a.m. To: [hidden email] Subject: Imaging Sourdough batter ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Colleagues, Here's something to chew on. This is not strictly a confocal question, but applies to widefield as well. Some folks here are interested in analysis of the time course of the maturation of sourdough, and have asked us to help them prepare samples of the batter for analysis of, say, yeast content. Have any of you had any experience with this sort of material? We have thought about making frozen sections of the batter at different times after initiation, and staining with DAPI. I'm curious about fixation (note that there are some interesting pH issues here), and quantitation methods. Any suggestions will be appreciated. Joel Joel B. Sheffield, Ph.D Department of Biology Temple University Philadelphia, PA 19122 Voice: 215 204 8839 e-mail: [hidden email] URL: *https://bio.cst.temple.edu/~jbs/ <https://bio.cst.temple.edu/%7Ejbs/> <http://tinyurl.com/khbouft>* |
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In reply to this post by JOEL B. SHEFFIELD
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Joel If you are intending to do this in time-lapse or real time I can set you up with a slick environmental control means where your folks can observe the process. I use this technique for live-yeast demonstrations in the classroom. Let me know what kind of scope you use, upright or inverted, and the objective magnification and I can send you some config notes. It will be easy! Dan On Mar 20, 2018, at 3:48 PM, Joel Sheffield wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Colleagues, Here's something to chew on. This is not strictly a confocal question, but applies to widefield as well. Some folks here are interested in analysis of the time course of the maturation of sourdough, and have asked us to help them prepare samples of the batter for analysis of, say, yeast content. Have any of you had any experience with this sort of material? We have thought about making frozen sections of the batter at different times after initiation, and staining with DAPI. I'm curious about fixation (note that there are some interesting pH issues here), and quantitation methods. Any suggestions will be appreciated. Joel Joel B. Sheffield, Ph.D Department of Biology Temple University Philadelphia, PA 19122 Voice: 215 204 8839 e-mail: [hidden email] URL: *https://bio.cst.temple.edu/~jbs/ <https://bio.cst.temple.edu/%7Ejbs/> <http://tinyurl.com/khbouft>* Dan Focht Bioptechs Inc. 3560 Beck Road Butler, PA 16002-9259 Office: 724-282-7145 Toll Free: 877-LIVE-CELL (548-3235) [hidden email] www.bioptechs.com |
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In reply to this post by JOEL B. SHEFFIELD
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Joel, In addition to the other replies: * near infrared fluorescent dye for DNA (nuclei) * Eosin and/or other "protein(s)" binding dye(s) ... no need to go for uniform labeling. Dough could be mostly unlabeled, some (highly) labeled stuff folded in. * nanoparticles and/or microparticles (an earlier reply mentioned Calcofluor yeast 'biomicroparticles'). * as mentioned in another reply: fluorescent protein expressing yeast etc. ("redder is better" for imaging deep). * autofluorescence ... especially with multiphoton excitation for depth and my #1 recommendation: * reflected light, preferably by confocal detection. Longer wavelength will enable greater depth. enjoy, George p.s. if that goes well, get (or make) a cryostage and make and image during making 'green (and red and NIR) fluorescent ice cream'. This p.s. inspired by My factoid calendar of the day, which today revealed that ice cream was invented in China over 2000 years ago. On 3/20/2018 3:48 PM, Joel Sheffield wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Colleagues, > > Here's something to chew on. > > This is not strictly a confocal question, but applies to widefield as > well. Some folks here are interested in analysis of the time course of the > maturation of sourdough, and have asked us to help them prepare samples of > the batter for analysis of, say, yeast content. Have any of you had any > experience with this sort of material? We have thought about making frozen > sections of the batter at different times after initiation, and staining > with DAPI. I'm curious about fixation (note that there are some interesting > pH issues here), and quantitation methods. Any suggestions will be > appreciated. > > Joel > > > Joel B. Sheffield, Ph.D > Department of Biology > Temple University > Philadelphia, PA 19122 > Voice: 215 204 8839 > e-mail: [hidden email] > URL: *https://bio.cst.temple.edu/~jbs/ > <https://bio.cst.temple.edu/%7Ejbs/> <http://tinyurl.com/khbouft>* -- George McNamara, PhD Baltimore, MD 21231 [hidden email] https://www.linkedin.com/in/georgemcnamara https://works.bepress.com/gmcnamara/75 (may need to use Microsoft Edge or Firefox, rather than Google Chrome) http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650 http://confocal.jhu.edu July 2017 Current Protocols article, open access: UNIT 4.4 Microscopy and Image Analysis http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/abstract supporting materials direct link is http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/full#hg0404-sec-0023 figures at http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/figures |
 
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Richard E. Edelmann |
Re: Imaging Sourdough batter
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In reply to this post by JOEL B. SHEFFIELD
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Joel, Not sure what you have access to, I have done several projects with mayonnaise, ice cream, and cookie dough. Primarily focusing on air and fat size and distribution. This is easily done via cryo-SEM (LTSEM). The low water content, the freezing point depression and high ice nucleation all make for easy freezing with fine crystals. Frozen fracture lets you look at the true matrix structure. But microbial identification would be by morphology unless you get creative with something like inter-cellular QDOTs. Good luck and let us know how it goes! On Tue, Mar 20, 2018 at 3:48 PM, Joel Sheffield <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Colleagues, > > Here's something to chew on. > > This is not strictly a confocal question, but applies to widefield as > well. Some folks here are interested in analysis of the time course of the > maturation of sourdough, and have asked us to help them prepare samples of > the batter for analysis of, say, yeast content. Have any of you had any > experience with this sort of material? We have thought about making frozen > sections of the batter at different times after initiation, and staining > with DAPI. I'm curious about fixation (note that there are some interesting > pH issues here), and quantitation methods. Any suggestions will be > appreciated. > > Joel > > > Joel B. Sheffield, Ph.D > Department of Biology > Temple University > Philadelphia, PA 19122 > Voice: 215 204 8839 > e-mail: [hidden email] > URL: *https://bio.cst.temple.edu/~jbs/ > <https://bio.cst.temple.edu/%7Ejbs/> <http://tinyurl.com/khbouft>* > -- Richard E. Edelmann, Ph.D. Center for Advanced Microscopy & Imaging, Director 9C Upham Hall Miami University Oxford, OH 45056 Phone: 513-529-5712 Email: [hidden email] Web: www.cami.muohio.edu |
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