hair
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Does anyone have suggestions for imaging hair? We have someone who
wants to get a cross section. Is there some fluorescent
label? I am having difficulty with the density of the hair (light
doesn't penetrate very far).
Cytometry Facility Huck Institute of the Life Sciences 319 Life Sciences Building Penn State University University Park, PA 16802 http://www.huck.psu.edu/facilities/cytometry-up/ 814-863-2762 |
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Hi Elaine,
A simple way to get hair cross sections is to pull some of the hair and some cotton fibres through a small hole in a metal plate, then cut across with a sharp razor blade on either side of the plate (which shouldn¹t be too thick...). Place on a slide, add immersion oil or water, coverslip and you¹ll see a good cross-seciton of the hairs. Your workshop should be able to drill some small holes (can't remember dimensions, but small...) in a small piece of metal sheeting. This was part of a zoology prac. a long time ago, comparing hair from different animals, including members of the class. cheers, Rosmary Dr Rosemary White CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia ph 61 2 6246 5475 fx 61 2 6246 5334 On 4/12/08 8:27 AM, "Elaine Kunze" <[hidden email]> wrote: > Does anyone have suggestions for imaging hair? We have someone who wants to > get a cross section. Is there some fluorescent label? I am having difficulty > with the density of the hair (light doesn't penetrate very far). > > Elaine Kunze > Cytometry Facility > Huck Institute of the Life Sciences > 319 Life Sciences Building > Penn State University > University Park, PA 16802 > http://www.huck.psu.edu/facilities/cytometry-up/ > <http://www.huck.psu.edu/facilities/cytometry-up/> 814-863-2762 |
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Hi Elaine,
While not a confocal technique, I have gotten beautiful images of hair in transmitted light DIC. The quality of the image doesn't deteriorate too badly as you focus through and the pigment granules are beautiful because they're highly birefringent. I've used objective lenses with magnifications as high as 100X for this. Your students may want to compare different hair colors against their bleached and/or dyed equivalents. It's also interesting to look at the differences between the brown and white hairs of familiar animals. It will be difficult to look at hair in cross section with transmitted light DIC unless you can cut very thin sections. Jeffrey Larson Product Manager Nikon Confocal Systems Nikon Instruments Inc. (631) 547-8540 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Rosemary White Sent: Wednesday, December 03, 2008 4:52 PM To: [hidden email] Subject: Re: hair Hi Elaine, A simple way to get hair cross sections is to pull some of the hair and some cotton fibres through a small hole in a metal plate, then cut across with a sharp razor blade on either side of the plate (which shouldn¹t be too thick...). Place on a slide, add immersion oil or water, coverslip and you¹ll see a good cross-seciton of the hairs. Your workshop should be able to drill some small holes (can't remember dimensions, but small...) in a small piece of metal sheeting. This was part of a zoology prac. a long time ago, comparing hair from different animals, including members of the class. cheers, Rosmary Dr Rosemary White CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia ph 61 2 6246 5475 fx 61 2 6246 5334 On 4/12/08 8:27 AM, "Elaine Kunze" <[hidden email]> wrote: > Does anyone have suggestions for imaging hair? We have someone who wants to > get a cross section. Is there some fluorescent label? I am having difficulty > with the density of the hair (light doesn't penetrate very far). > > Elaine Kunze > Cytometry Facility > Huck Institute of the Life Sciences > 319 Life Sciences Building > Penn State University > University Park, PA 16802 > http://www.huck.psu.edu/facilities/cytometry-up/ > <http://www.huck.psu.edu/facilities/cytometry-up/> 814-863-2762 |
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In reply to this post by Elaine Kunze
Dear Elaine,
vital stains will work in the region of the hair bulb where you have some live cells. Incubate the freshly plucked hair with labels diluted in culture medium, rinse, then fix with paraformaldehyde. Some general protein stain such as eosin might be helpful in the acellular shaft. Your bigger problem will be the spherical aberration arising from refractive index of the hair. You may need to try different clearing agents such as glycerol dilutions, immersion oil or even higher RI reagents. Regards, Glen On Dec 3, 2008, at 1:27 PM, Elaine Kunze wrote: > Does anyone have suggestions for imaging hair? We have someone who > wants to get a cross section. Is there some fluorescent label? I > am having difficulty with the density of the hair (light doesn't > penetrate very far). > Elaine Kunze > Cytometry Facility > Huck Institute of the Life Sciences > 319 Life Sciences Building > Penn State University > University Park, PA 16802 > http://www.huck.psu.edu/facilities/cytometry-up/ > 814-863-2762 > Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 [hidden email] ****************************************************************************** The box said "Requires WindowsXP or better", so I bought a Macintosh. ****************************************************************************** |
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Hi Elaine,
TDE treatment (ending in 96% TDE) seems to work well with hair follicles; I imaged these with 20x oil and 40x oil on a Leica SP5 and was able to see through the bulb. The hair shafts that I used were autofluorescent at 561 nm excitation with a rather spiky emission spectrum (anyone knows where this arises from?), giving great images even w/o staining. I can confirm that DIC Z stacks look great as well, I used the 561 and/or the 488 lines to record this on the SP5. A bit of deconvolution with theoretical PSF improves the DIC stacks even further.
Best, Zoltan On Wed, Dec 3, 2008 at 10:54 PM, Glen MacDonald <[hidden email]> wrote: Dear Elaine, -- Zoltan Cseresnyes Facility manager, Imaging Suite University of Cambridge, UK
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In reply to this post by Larson Jeffrey M.
I agree that DIC can work well for hair imaging, and it's an
excellent complementary technique for SEM of hair. There are some examples on the Michigan Mammals hair ID pages: http://www.cst.cmich.edu/users/swans1bj/Hair%20ID/Species.html Phil >Hi Elaine, > >While not a confocal technique, I have gotten beautiful images of >hair in transmitted light DIC. The quality of the image doesn't >deteriorate too badly as you focus through and the pigment granules >are beautiful because they're highly birefringent. I've used >objective lenses with magnifications as high as 100X for this. Your >students may want to compare different hair colors against their >bleached and/or dyed equivalents. It's also interesting to look at >the differences between the brown and white hairs of familiar >animals. It will be difficult to look at hair in cross section with >transmitted light DIC unless you can cut very thin sections. > >Jeffrey Larson >Product Manager >Nikon Confocal Systems >Nikon Instruments Inc. >(631) 547-8540 > >-----Original Message----- >From: Confocal Microscopy List >[mailto:[hidden email]] On Behalf Of Rosemary White >Sent: Wednesday, December 03, 2008 4:52 PM >To: [hidden email] >Subject: Re: hair > >Hi Elaine, > >A simple way to get hair cross sections is to pull some of the hair and some >cotton fibres through a small hole in a metal plate, then cut across with a >sharp razor blade on either side of the plate (which shouldn't be too >thick...). Place on a slide, add immersion oil or water, coverslip and >you'll see a good cross-seciton of the hairs. Your workshop should be able >to drill some small holes (can't remember dimensions, but small...) in a >small piece of metal sheeting. > >This was part of a zoology prac. a long time ago, comparing hair from >different animals, including members of the class. > >cheers, >Rosmary > >Dr Rosemary White >CSIRO Plant Industry >GPO Box 1600 >Canberra, ACT 2601 >Australia > >ph 61 2 6246 5475 >fx 61 2 6246 5334 > > > >On 4/12/08 8:27 AM, "Elaine Kunze" <[hidden email]> wrote: > >> Does anyone have suggestions for imaging hair? We have someone who wants to >> get a cross section. Is there some fluorescent label? I am >>having difficulty >> with the density of the hair (light doesn't penetrate very far). >> >> Elaine Kunze >> Cytometry Facility >> Huck Institute of the Life Sciences >> 319 Life Sciences Building >> Penn State University >> University Park, PA 16802 >> http://www.huck.psu.edu/facilities/cytometry-up/ >> <http://www.huck.psu.edu/facilities/cytometry-up/> 814-863-2762 -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 |
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In reply to this post by Elaine Kunze
Hii
Just try to SEM and a simple surgical blade to section hair,
Let me know teh purpose i will be able to suggest you something
I work a lot on hair cross sections and Imaging hair bulbs etc
Rgds
On Thu, Dec 4, 2008 at 2:57 AM, Elaine Kunze <[hidden email]> wrote:
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