Aloe-leaf epidermal peel - stomata
Aloe-leaf epidermal peel - stomata
Hi all, I've been putting some freshly-cut Aloe-leaf into fixative this evening and thought I'd make a quick & dirty epidermal-peel. Stained briefly with Toluidine-blue (aq) then water-mounted with coverslip, very colourful.
Had a quick try of a cardboard 'Matthias-arrow' I 'made' when reading the great Walter Dioni's articles when I started-out - it's really not very good compared to the simply fantastic oblique-illumination often seen of this forum (to say the least! ) but gives a little relief at least.
Here are a few pictures, apologies for the poor quality, it was a very quick foray as I was actually fixing tissue rather than meaning to do this, but though it'd be nice to have a peek. I'll definitely go back and do some 'proper' ones I think, maybe mounted in glycerin or perhaps an alcohol-based mountant, anyway here are a few pictures to peruse!
A real 'purple-haze' and closer-in, more, and a rather tragic attempt at oblique by way of a handy Matthias-arrow.. That's all I have, good fun having a go though, must do some more live-tissue when I get a chance.
To sleep for me, back soon..
Had a quick try of a cardboard 'Matthias-arrow' I 'made' when reading the great Walter Dioni's articles when I started-out - it's really not very good compared to the simply fantastic oblique-illumination often seen of this forum (to say the least! ) but gives a little relief at least.
Here are a few pictures, apologies for the poor quality, it was a very quick foray as I was actually fixing tissue rather than meaning to do this, but though it'd be nice to have a peek. I'll definitely go back and do some 'proper' ones I think, maybe mounted in glycerin or perhaps an alcohol-based mountant, anyway here are a few pictures to peruse!
A real 'purple-haze' and closer-in, more, and a rather tragic attempt at oblique by way of a handy Matthias-arrow.. That's all I have, good fun having a go though, must do some more live-tissue when I get a chance.
To sleep for me, back soon..
John B
Re: Aloe-leaf epidermal peel - stomata
Very beautiful, John, and absolute image clarity.
Re: Aloe-leaf epidermal peel - stomata
I agree with Gekko,beautiful pictures.
-Reichert Polyvar
-Olympus IX70
-Zeiss Photomicroscope
-Canon 600D
-Olympus IX70
-Zeiss Photomicroscope
-Canon 600D
Re: Aloe-leaf epidermal peel - stomata
Hey John,
Not seeing the poor quality you are apologizing for, great images!
This is more the approach that I see myself undertaking, great to see such excellent results are possible.
Any tips or links on staining specimens under this protocol? I am ham-fisted in my efforts with staining.
Great stuff as usual!
Rod
Not seeing the poor quality you are apologizing for, great images!
This is more the approach that I see myself undertaking, great to see such excellent results are possible.
Any tips or links on staining specimens under this protocol? I am ham-fisted in my efforts with staining.
Great stuff as usual!
Rod
Re: Aloe-leaf epidermal peel - stomata
Thanks Rod! I buy my Toluidine-blue stain, the very best stain for living (e.g. hand-cut) tissue (plant) in my experience, from this e-bayer in the UK for what is frankly a ridiculously low price http://www.ebay.co.uk/itm/Histology-Sta ... jx9U6JCThwrnabholz wrote:Hey John,
Not seeing the poor quality you are apologizing for, great images!
This is more the approach that I see myself undertaking, great to see such excellent results are possible.
Any tips or links on staining specimens under this protocol? I am ham-fisted in my efforts with staining.
Great stuff as usual!
Rod
In the USA perhaps this one would be OK? http://www.ebay.co.uk/itm/EK-Industries ... Sw3xJVZ4Hp
I buy the 0.5% aqueous mix and dilute with water (de-ionised is best) to give my working-solution of 0.05% aqueous. This strength is for my tissue fixed to slides and left in the stain, for 'on-the-fly' staining I use 0.5% from a dropper-bottle straight onto the tissue. It's a superb stain for live tissues and simplicity to use.
Dropper plenty (of the 0.5%) onto your tissue, on a slide or in a dish etc and let the stain act, probably for about 2 minutes, vary as required. When you think it's right, rinse with water to remove excess stain, look at the staining with a 'scope and stain further if necessary or rinse in water or OH to remove some stain (easier just to run a series of tests than to remove over-staining though).
This stain will act metachromatically and usually gives beautiful rich blues, pinks, reds etc as partly seen in my pictures. A brief and gentle rinse in tap-water often adds a little intensity to a stain once at the desired level of staining. I think the alkalinity causes an effect known as 'bluing' - certainly works for me.
Best then to coverslip for a good water-mount, keeps everything flat and stable!
Go to it, you'll really enjoy the results. Have a look at some of my posts re hand-sections, the stain is demonstrated in them with some really beautiful results. Hand-sections can be just as rewarding and beautiful as the full-blown permanent sections that I make, especially now we can photograph them to such high standards for permanent record!
Have fun!
Last edited by mrsonchus on Mon Jan 11, 2016 9:58 pm, edited 1 time in total.
John B
Re: Aloe-leaf epidermal peel - stomata
Thanks Seb & Gekko, you always bring a smile to my face with your kind comments. Pleased you like them.
John B
Re: Aloe-leaf epidermal peel - stomata
Stain definitely brings out the cell contents. Beautiful images and detail. I like live cell sections the best.
JimT
JimT
Re: Aloe-leaf epidermal peel - stomata
You're right there Jim, there's an inherent beauty in the live tissue that I never really see in the processed sections. For sheer detail and information I think the 'full-process' sections are superior, but for anadulterated beauty and a real sense of the plant whence came the tissue, the live sections always come out on top for me.JimT wrote:Stain definitely brings out the cell contents. Beautiful images and detail. I like live cell sections the best.
JimT
Thanks chaps for your compliments, I'm very pleased you like these.
John B
Re: Aloe-leaf epidermal peel - stomata
Thanks for the run down on staining John.
I need to quit my job to have enough time to do all of this cool stuff.
Maybe the Powerball Lottery is the answer, $1billion this week... That would buy a lot of stain.
Thanks
Rod
I need to quit my job to have enough time to do all of this cool stuff.
Maybe the Powerball Lottery is the answer, $1billion this week... That would buy a lot of stain.
Thanks
Rod
Re: Aloe-leaf epidermal peel - stomata
You just convinced me to order some. You had recommended it before but I went for methylene blue instead. I like it but will give this a try.
JimT
JimT
Re: Aloe-leaf epidermal peel - stomata
Go get it Jim - it's definitely superior to Methylene blue, although for processed tissue Methylene blue is capable of very fine detail, it can for some reason cause quite a brutal 'tattering' of some tissue - no idea why though..JimT wrote:You just convinced me to order some. You had recommended it before but I went for methylene blue instead. I like it but will give this a try.
JimT
You'll see in the pictures that the Toluidine blue will give a very nice (blue) nuclear image too, and doesn't tend to simply 'black-out' the details as a lot of stains often do with nuclei.
Toluidine blue is superb for live tissue, nothing I've tried comes close, although second place is undoubtedly occupied by Safranin I find.
[
If you find your aqueous stains are having trouble penetrating some tissue or areas within, a little OH can often remove certain hydrophobic molecules and help with staining.. Rehydrate the tissue after the OH wash and before staining for best results.
]
Good luck!
John B
Re: Aloe-leaf epidermal peel - stomata
My Toluidine-blue is on the way too.
Thanks for the direction John.
Rod
Thanks for the direction John.
Rod
Re: Aloe-leaf epidermal peel - stomata
You'll be pleased with it for certain chaps, fun is coming your way!
You've probably seen these from my earlier posts re hand-sectioning etc but I thought I'd show what Methylene-blue and Safranin can do with live tissue (used separately that is, not combined)..
Here's a hand-sectioned, Methylene-blue stained vascular bundle, epidermis, cortex and parenchyma-cells; Here's the hand-sectioned central vascular-cylinder from a grass-stem stained with Safranin - shows several different kinds of vessel-wall structures.. This is a hand-sectioned (chrysanthemum-stem I think...) vascular bundle tri-stained with Crystal-violet, Safranin and I think Fast-green (although I'm not certain of that one.. ) Finally here's a hand-sectioned stem rather brutally overstained by me with Toluidine-blue, when I was somewhat 'less experienced... ' than I (hopefully) am now - subtlety seems to have eluded me here! Some metachromasia though! Apologies if you've seen these before, just though I'd put them up in case they're new to you.
All were done during my early-days and were just a case of the simple protocol mentioned earlier - a little practice and you're off and running!
Staining is a black-art - and superbly rewarding when it goes well - as it always does after a little practice!
Good luck chaps!
You've probably seen these from my earlier posts re hand-sectioning etc but I thought I'd show what Methylene-blue and Safranin can do with live tissue (used separately that is, not combined)..
Here's a hand-sectioned, Methylene-blue stained vascular bundle, epidermis, cortex and parenchyma-cells; Here's the hand-sectioned central vascular-cylinder from a grass-stem stained with Safranin - shows several different kinds of vessel-wall structures.. This is a hand-sectioned (chrysanthemum-stem I think...) vascular bundle tri-stained with Crystal-violet, Safranin and I think Fast-green (although I'm not certain of that one.. ) Finally here's a hand-sectioned stem rather brutally overstained by me with Toluidine-blue, when I was somewhat 'less experienced... ' than I (hopefully) am now - subtlety seems to have eluded me here! Some metachromasia though! Apologies if you've seen these before, just though I'd put them up in case they're new to you.
All were done during my early-days and were just a case of the simple protocol mentioned earlier - a little practice and you're off and running!
Staining is a black-art - and superbly rewarding when it goes well - as it always does after a little practice!
Good luck chaps!
John B
Re: Aloe-leaf epidermal peel - stomata
Bonjour
Les photos sont très belles et très bien expliquer.
Merci pour le partage
Cordialement seb
Les photos sont très belles et très bien expliquer.
Merci pour le partage
Cordialement seb
Microscope Leitz Laborlux k
Boitier EOS 1200D + EOS 1100D
Boitier EOS 1200D + EOS 1100D