F R e N c H 3 z Posted November 15, 2010 Report Share Posted November 15, 2010 Here are a few attempts at examining some CPs under a confocal microscope. As firsts I can't complain though I wish I would have changed a few things now. The digestive gland at the very bottom of the first picture was deeper than the rest of the plane which resulted in the unfortunate large dark shadow at the bottom half of the page. However due to the nice deep red color of the gland the ones that did come out did so quite nicely. Would attempt some TEM/SEM work with these but the time requirement to do so just isn't feasible. Scaling will also have to be adjusted in order to be more visible. Dionaea muscipula digestive glands Dionaea muscipula trigger hair base Playing with contrast options, Pinguicula pedunculate and sessile glands Drosera multifida tentacle Contrast/brightness played with Nepenthes digestive gland Stylidium debile petal, pigment shown is the dark purple striping across each petal Not plant related; Mouse kidney tissue under TEM; shown are cristae within mitochondria Thanks for looking! Quote Link to comment Share on other sites More sharing options...
vraev Posted November 15, 2010 Report Share Posted November 15, 2010 Really amazing. My supervisor doesn't allow us to use all these facilities even though we have them available. I guess it is for a good reason... Per hour in the confocal facility costs us like a 150$ or so. Quote Link to comment Share on other sites More sharing options...
mantrid Posted November 15, 2010 Report Share Posted November 15, 2010 Some excellent shots. I would like to see some TEM cross sections through Dionaea digestive gland. Quote Link to comment Share on other sites More sharing options...
F R e N c H 3 z Posted November 15, 2010 Author Report Share Posted November 15, 2010 Thank you, sorry to hear your supervisor restricts the use of yours. Some of those shots actually took about 2 hours so the process can get quite time consuming, and if you're looking to increase the quality you can easily be looking 3-4 hours to scan. The shots look cool because of the emission colors but the future of microscopy lays in EM. These are far from professional shots but I figured at least 1 or 2 people would be interested in close up views! As much as I would LOVE to do some SEM or even TEM work with the plants, the prep time to get a sample ready usually takes anywhere from 1-2 weeks. And if 1 procedure didn't go exactly right, you won't know until your sample is being viewed, then you realize you've wasted 2 weeks... Too time consuming at the moment between classes, projects and a thesis... Hopefully someday... Quote Link to comment Share on other sites More sharing options...
mantrid Posted November 16, 2010 Report Share Posted November 16, 2010 (edited) Thank you, sorry to hear your supervisor restricts the use of yours. Some of those shots actually took about 2 hours so the process can get quite time consuming, and if you're looking to increase the quality you can easily be looking 3-4 hours to scan. The shots look cool because of the emission colors but the future of microscopy lays in EM. These are far from professional shots but I figured at least 1 or 2 people would be interested in close up views!As much as I would LOVE to do some SEM or even TEM work with the plants, the prep time to get a sample ready usually takes anywhere from 1-2 weeks. And if 1 procedure didn't go exactly right, you won't know until your sample is being viewed, then you realize you've wasted 2 weeks... Too time consuming at the moment between classes, projects and a thesis... Hopefully someday... When I had something non project related I wanted to look at I did the prep alongside my project stuff. Nobody knew and it didnt take too much extra time. It was a very large EM suite and I pretty much had the run of the place, my supervisor was hardly ever around so nobody looking over my shoulder asking what I was working on. Im out of the field now so dont have access to such things anymore. Edited November 16, 2010 by mantrid Quote Link to comment Share on other sites More sharing options...
F R e N c H 3 z Posted November 16, 2010 Author Report Share Posted November 16, 2010 Wow I gotta I'm a bit jealous :) Have any shots to share? ;) Our lab time is by appointment only and the supervisors have insane schedules of their own : / I do have a couple more TEM shots from mouse lung tissue but nothing plant related unfortunately... Quote Link to comment Share on other sites More sharing options...
jimfoxy Posted November 16, 2010 Report Share Posted November 16, 2010 Fascinating shots - thank you for taking the time to show them. Quote Link to comment Share on other sites More sharing options...
mantrid Posted November 16, 2010 Report Share Posted November 16, 2010 Wow I gotta I'm a bit jealous :) Have any shots to share? ;)Our lab time is by appointment only and the supervisors have insane schedules of their own : / I do have a couple more TEM shots from mouse lung tissue but nothing plant related unfortunately... I used to have boxes of photographs, digital wasnt used then. I got rid of most as they were taking up too much space. I probably still got the good/important ones in the attic somewhere. If I ever have time to find and scan them Ill post some. The TEM wont mean much to the average person but the SEM are pretty impressive. Quote Link to comment Share on other sites More sharing options...
F R e N c H 3 z Posted November 16, 2010 Author Report Share Posted November 16, 2010 My pleasure Jimfoxy :) @ Mantrid: Both are SEM and TEMs are still on polaroid. We haven't been digitalized yet either. The above shot was actually a scanned polaroid that had the brightness and contrast played with on photoshop. Look forward to seeing your shot when you get a chance. Quote Link to comment Share on other sites More sharing options...
Daniel O. Posted November 16, 2010 Report Share Posted November 16, 2010 Wow, really interesting shots. Best regards, Dani Quote Link to comment Share on other sites More sharing options...
F R e N c H 3 z Posted December 10, 2010 Author Report Share Posted December 10, 2010 Scale is hard to read on those, my apologies. Reads 100μm on most. Sphagnum hyaline cells: Sarracenia seedling lid. Hood hairs (Zone 1) can be slightly distinguished up top though they were hard to get a picture of due to their transparent appearances. At the just the right angle however they can be captured. The lighter area represent white pigmentation while darker pigmentation are actually redish pigmentation: Sarracenia seedling zone 4 trichome hairs. Much more distinguishable than zone 1 hairs. Image was taken from the same pitcher as the above shot. Pinguicula glandular and sessile glands: Purple pigmentation on Stylidium debile petal: Mouse lung tissue, empty spaces are bronchioles *I believe*. (Don't have a histology book handy) Quote Link to comment Share on other sites More sharing options...
Vic2 Posted December 10, 2010 Report Share Posted December 10, 2010 Nice shots! The mouse lung tissue seems to be loaded with mitochondria - I wasn't expecting that. Isn't oxygen transfer across the lungs a passive diffusive process? To add to mantrid's wish list, I'd like to see a TEM section through a Dionaea trap hinge. It's capable of the fastest growth in any Kingdom, apparently... And - again like mantrid - I found the time to embed my Mum's favourite four-leaf clover in TAAB EM resin, while I was sectioning honeybee Nasanov glands, and gave my delighted parent a personalised pendant necklace for Xmas... Well, I was student, and any spare cash was needed for essential liquid nutrition Vic Quote Link to comment Share on other sites More sharing options...
F R e N c H 3 z Posted December 10, 2010 Author Report Share Posted December 10, 2010 Thank you for the kind words. I was not expecting the amount of mitochondria present either. It seems that as if they are extracellular somehow as no cells can be seen let alone a nucleii. Perhaps something occurred during staining. The teaching staff for some reason decided against embedding plant materials. How did you go about the dehydration process since there would be cell walls to take into account? These mouse cells were treated with glutalraldehyde and imbedded with Osmium tertroxide. Will try to find out the procedure and chemicals for plant material. Personally I much prefer the confocal to the TEM, however had our SEM not been down all semester I totally would have been all over it! Quote Link to comment Share on other sites More sharing options...
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