Starch-stearic acid
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I have a
student that wants to image starch-paste and stearic acid. The aim is to
show the starch granules coated with stearic acid. We have tried FITC to
stain the starch, but that gives a very high background. For the stearic
acid we used All
suggestions and comments will be appreciated. TIA Alan Hall Laboratory for Microscopy & Microanalysis Room1-33, Natural Sciences 2 0002 Tel: +27 12 4202075 Fax: +27 12 3625150 |
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We were imaging amyloplasts (on confocal) isolated from fresh tissue using FM 4-64 to detect intact membranes, quite easy to do, you don’t really have to stain the starch with anything. They were very distinct in the transmitted light image. cheers, Rosemary p.s. go Crusaders! Rosemary White [hidden email] CSIRO Plant Industry ph. 61 (0)2-6246 5475 GPO Box 1600 fax. 61 (0)2-6246 5334 Canberra, ACT 2601 Australia On 30/5/08 5:54 PM, "Alan Hall" <[hidden email]> wrote: Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal |
 
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Re: Starch-stearic acid
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In reply to this post by Alan Hall
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I think Feulgen will turn starch a purplish color. It was a complication we found, when students in my course were trying to stain DNA. I think you could find a reference in old books on histology technique. Carol Heckman Bowling Green State University |
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In reply to this post by Rosemary.White
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For that matter, startch grains show up unmistakably in polarized
light. And if the stearic acid binds in an orderly way to the starch,
who knows, you might get a distinct ring-around-the-grain
effect!
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Why not just stain the stearic acid with nile red and don't worry about the starch. You will be able to see the starch grains in the transmitted light image very easily. --
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In reply to this post by Alan Hall
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If you have acces to a 2-photon system, starch gives a very stong second harmonic signal. See Guy Cox, Nuno Moreno & José Feijó, 2005. Second harmonic imaging of plant polysaccharides. Journal of Biomedical Optics 10, 024013_1-6
Guy Optical Imaging Techniques in Cell Biology
by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net From: Confocal Microscopy List on behalf of Alan Hall Sent: Fri 08/05/30 5:54 PM To: [hidden email] Subject: Starch-stearic acid Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
I have a student that wants to image starch-paste and stearic acid. The aim is to show the starch granules coated with stearic acid. We have tried FITC to stain the starch, but that gives a very high background. For the stearic acid we used Nile red.
All suggestions and comments will be appreciated. TIA Alan Hall
Laboratory for Microscopy & Microanalysis Room1-33, Natural Sciences 2 University of Pretoria Lynnwood Road Pretoria 0002 Tel: +27 12 4202075 Fax: +27 12 3625150 Mobile: +27 82 3319040
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In reply to this post by Alan Hall
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I have tried several techniques to try and get good images
from starch and other polysaccahrides within food samples. If the starch
grains are intact or even fairly intact they can show up using transmission mode
as the mebranes are still visible but it is not that easy to see them in some
cases. I have had some success with lectin probes but it is a bit variable
depending on other substances in the sample. I am not sure with your case. And
there is a feulgens method which you can use if the treatment is not going to
affect your sample as you have to hydrolyse first.
Using polarised light only seems to work well when your
sample is well dispersed and the grains still intact, it is very hard to see
them in denser samples. From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Alan Hall Sent: Friday, 30 May 2008 7:55 p.m. To: [hidden email] Subject: Starch-stearic acid I have a student that
wants to image starch-paste and stearic acid. The aim is to show the
starch granules coated with stearic acid. We have tried FITC to stain the
starch, but that gives a very high background. For the stearic acid we
used All suggestions and
comments will be appreciated. TIA Alan Hall
Laboratory for Microscopy &
Microanalysis Room1-33, Natural Sciences
2 0002 Tel: +27 12
4202075 Fax: +27 12
3625150 DISCLAIMER: |
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