Botanic section from paraffin block with unreadable label
Botanic section from paraffin block with unreadable label
Hi together,
I came to a couple of paraffin blocks with unreadable or missing labels.
Here images of the block and a cross section through it. It is actually quite attractive. 10µ sections stained in Alcian blue - Rhodamine - Acriflavine 4:1:1 for 7 minutes cold.
Anybody an idea what this might be. Label sais "Cherwieph."???
Bob
I came to a couple of paraffin blocks with unreadable or missing labels.
Here images of the block and a cross section through it. It is actually quite attractive. 10µ sections stained in Alcian blue - Rhodamine - Acriflavine 4:1:1 for 7 minutes cold.
Anybody an idea what this might be. Label sais "Cherwieph."???
Bob
- Attachments
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- Cherwieph. Foto Block 1024.jpg (157.15 KiB) Viewed 4853 times
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- Cherwieph. 1024.jpg (93.49 KiB) Viewed 4853 times
Re: Botanic section from paraffin block with unreadable label
Aha! Definitely a moss-leaf I'd say. Not sure of an ID though. It's similar to a couple of hand-sections I've taken in the past link to images
John B
Re: Botanic section from paraffin block with unreadable label
Aha! Definitely a moss-leaf I'd say. Not sure of an ID though. It's similar to a couple of hand-sections I've taken in the past link to images
The block looks to contain a complete moss 'branch' or shoot (perpendicular and 'top-first' to the knife's-cutting plane), that has a stem in the centre (yet to be reached by the knife) and leaves around. You appear to have TSs of some leaves at this level into the block. If you keep-going deeper into this block, checking very carefully after say every 100µ or-so, you may be lucky and make a few sections across the actual growing-tip (AKA 'shoot meristem').
If this is the case, you'll likely come first across a large triangular (in TS) single apical cell, which as it's passed by and through with subsequent sections will be surrounded by more cells, comprising the 'meristem' proper. I think in mosses the apical cell, whilst being the 'mother of all cells' in moss, divides infrequently - further more frequent divisions being from the immediately-surrounding 'daughter cells' (AKA 'initials') that are cut-off from each of the 3 faces of the apical cell - i.e. in whorls of 3 at each level.
This should be a very interesting block to 'dig into' - but you'll need to check constantly and try to section at about 5µ to catch the 'sweet spot' which is the meristem.
Looking forward to more!
The block looks to contain a complete moss 'branch' or shoot (perpendicular and 'top-first' to the knife's-cutting plane), that has a stem in the centre (yet to be reached by the knife) and leaves around. You appear to have TSs of some leaves at this level into the block. If you keep-going deeper into this block, checking very carefully after say every 100µ or-so, you may be lucky and make a few sections across the actual growing-tip (AKA 'shoot meristem').
If this is the case, you'll likely come first across a large triangular (in TS) single apical cell, which as it's passed by and through with subsequent sections will be surrounded by more cells, comprising the 'meristem' proper. I think in mosses the apical cell, whilst being the 'mother of all cells' in moss, divides infrequently - further more frequent divisions being from the immediately-surrounding 'daughter cells' (AKA 'initials') that are cut-off from each of the 3 faces of the apical cell - i.e. in whorls of 3 at each level.
This should be a very interesting block to 'dig into' - but you'll need to check constantly and try to section at about 5µ to catch the 'sweet spot' which is the meristem.
Looking forward to more!
John B
Re: Botanic section from paraffin block with unreadable label
Hi John,
thank you for your identification! It really is a moss-leaf. This also fits to the overall look of the plant object. The question is now what "Cherwieph." might mean. It is old hand writing and could either be german or latin, also an abreviation or part of a name. Simple task!
The paraffin it is embedded in is very brittle unfortunately. I have knifes with more acute angles and might try it with them. Knowing that this is a moss I might try to embed mosses myself as this is just the kind of object I like to visualize.
Bob
thank you for your identification! It really is a moss-leaf. This also fits to the overall look of the plant object. The question is now what "Cherwieph." might mean. It is old hand writing and could either be german or latin, also an abreviation or part of a name. Simple task!
The paraffin it is embedded in is very brittle unfortunately. I have knifes with more acute angles and might try it with them. Knowing that this is a moss I might try to embed mosses myself as this is just the kind of object I like to visualize.
Bob
- Attachments
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- Cherwieph..jpg (23.05 KiB) Viewed 4840 times
Re: Botanic section from paraffin block with unreadable label
Bob,
You might try re-heating the block to say 45 (and 50 etc) Centigrade and see whether the wax resets to a more easy state to process.
Best,
deBult
You might try re-heating the block to say 45 (and 50 etc) Centigrade and see whether the wax resets to a more easy state to process.
Best,
deBult
Re: Botanic section from paraffin block with unreadable label
Hi Bob - you've been nibbled by the 'sectioning-bug'!!
As suggested above, I would go a step further though. Put the block into some molten wax of the kind you're using and wait for it to melt completely, at which time the moss will become visible in the liquid wax. Then, simply pass the shoot through say 3 firther baths of new (molten) wax (perhaps 3 hours each stage). This should be enough to displace/replace the old wax throughout the tissue and enable a fresh re-cast of the moss shoot tobe made. This technique definitely works.
When I go to the embedding phase with my tissue, which will of course be in it's final infiltration wax, I cast as many blocks of the tissue I desire, and am invariably left with lots of pieces in the infiltration wax that are simply 'too good to be discarded'. That's where I cast one or two of what I call 'bulk blocks'. I simply dump all the remaining pieces into large blocks and let them set in the usual way, no orientation - just storage.
At a later time I will re-embed (the tissue is already thoroughly infiltrated with wax) and reorient of course for the cut desired, into new blocks.
With your old block you just need to add the extra wax baths as you are going to re-infiltrate the tissue (moss) as well as re-cast the block....
Aha - found some images of bulk-blocks for re-casting from slide making at the beginning of this month, link to dandelion root bulk block and link to coleus bulk block
and link to re-embedded sonchus stem tissue
The coleus tissue, initially cast in 58 deg wax, was not sectioning very well, so I re-infiltrated and re-cast it into 52 deg 'Paraplast +' - from which it has sectioned very nicely. Don't be afraid to re-cast or re-infiltrate and cast - it can save what you may have thought was a poor tissue, or be very good if you need to re-orientate for better sectioning planes.
As suggested above, I would go a step further though. Put the block into some molten wax of the kind you're using and wait for it to melt completely, at which time the moss will become visible in the liquid wax. Then, simply pass the shoot through say 3 firther baths of new (molten) wax (perhaps 3 hours each stage). This should be enough to displace/replace the old wax throughout the tissue and enable a fresh re-cast of the moss shoot tobe made. This technique definitely works.
When I go to the embedding phase with my tissue, which will of course be in it's final infiltration wax, I cast as many blocks of the tissue I desire, and am invariably left with lots of pieces in the infiltration wax that are simply 'too good to be discarded'. That's where I cast one or two of what I call 'bulk blocks'. I simply dump all the remaining pieces into large blocks and let them set in the usual way, no orientation - just storage.
At a later time I will re-embed (the tissue is already thoroughly infiltrated with wax) and reorient of course for the cut desired, into new blocks.
With your old block you just need to add the extra wax baths as you are going to re-infiltrate the tissue (moss) as well as re-cast the block....
Aha - found some images of bulk-blocks for re-casting from slide making at the beginning of this month, link to dandelion root bulk block and link to coleus bulk block
and link to re-embedded sonchus stem tissue
The coleus tissue, initially cast in 58 deg wax, was not sectioning very well, so I re-infiltrated and re-cast it into 52 deg 'Paraplast +' - from which it has sectioned very nicely. Don't be afraid to re-cast or re-infiltrate and cast - it can save what you may have thought was a poor tissue, or be very good if you need to re-orientate for better sectioning planes.
John B
Re: Botanic section from paraffin block with unreadable label
Hi Maarten,
I tried to soften the paraffin by warming it, this section was the result. But the block didn't behave well for more than a few sections.
So I think exchangeing the paraffin is the better way.
@John: Thank you for the pointer towards the re-embedding! This is probably easy to do as a I have little jars with some of the rest of 60°C parffin ready, that I can use up for this.
The new paraffin I got is Paraplast + with DMSO. This must not be heated above 62°C so I have to take care not to spoil it. For heating I have used a hot plate that I can cover with a glass vase. I think about investing in a cooking pot to get a nicer closed warm area for this. I remember you used a tuned pizza oven but have changed to something else lately. What can you recommend for heating purposes?
Bob
I tried to soften the paraffin by warming it, this section was the result. But the block didn't behave well for more than a few sections.
So I think exchangeing the paraffin is the better way.
@John: Thank you for the pointer towards the re-embedding! This is probably easy to do as a I have little jars with some of the rest of 60°C parffin ready, that I can use up for this.
The new paraffin I got is Paraplast + with DMSO. This must not be heated above 62°C so I have to take care not to spoil it. For heating I have used a hot plate that I can cover with a glass vase. I think about investing in a cooking pot to get a nicer closed warm area for this. I remember you used a tuned pizza oven but have changed to something else lately. What can you recommend for heating purposes?
Bob
Re: Botanic section from paraffin block with unreadable label
Hi Bob, I still use the little oven, controlled by a separate PID thermo-switch, with the oven's own thermostat as the backup.
I also now use a 'dry-block' heater, which gives very accurate temperature control indeed - in fact it's all about the max-min curve response of the heater - the shallower the curve the less the over/undershoot of the response of any thermo-switch.
Here's a link to my post with the dry-block heater used... - cost me about £30 on e-bay as I remember! As you can see, the holes in the aluminium blocks are really useful for keeping tools warm - essential for good orientation of tissue.
Here's a link to my post with the mini-oven...
The dry-block is used for embedding, the oven is used for infiltration.
I also now use a 'dry-block' heater, which gives very accurate temperature control indeed - in fact it's all about the max-min curve response of the heater - the shallower the curve the less the over/undershoot of the response of any thermo-switch.
Here's a link to my post with the dry-block heater used... - cost me about £30 on e-bay as I remember! As you can see, the holes in the aluminium blocks are really useful for keeping tools warm - essential for good orientation of tissue.
Here's a link to my post with the mini-oven...
The dry-block is used for embedding, the oven is used for infiltration.
John B
Re: Botanic section from paraffin block with unreadable label
Hi John,
thank you for the pointers towards the equipment you use!
The melting and handling of molten paraffin was a bit difficult for me. For the melting I used the affordable hot plate I showed here: viewtopic.php?f=10&t=8102&p=71108&hilit=oven#p71108
With a rectangular glass vase as a cover. This worked quite well for infiltrating. But when I started embedding it showed that I hadnn't prepared my work well enough, forceps cold, cooling of the block not well prepared, paraffin becoming cold too quickly, paraffin bits everywhere.... This is the area where I have to prepare better solutions.
I have now bought a cast aluminium cooking pot to which I have applied a layer of heat resistant silicone to get best possible heat transmision. This should give a warm chamber and even has a glass lid.
A practical question about your oven: These have a timer - do you have to activate it every 60 minutes or does it heat continuously?
That double temperature shut-off is a very good solution!
Bob
thank you for the pointers towards the equipment you use!
The melting and handling of molten paraffin was a bit difficult for me. For the melting I used the affordable hot plate I showed here: viewtopic.php?f=10&t=8102&p=71108&hilit=oven#p71108
With a rectangular glass vase as a cover. This worked quite well for infiltrating. But when I started embedding it showed that I hadnn't prepared my work well enough, forceps cold, cooling of the block not well prepared, paraffin becoming cold too quickly, paraffin bits everywhere.... This is the area where I have to prepare better solutions.
I have now bought a cast aluminium cooking pot to which I have applied a layer of heat resistant silicone to get best possible heat transmision. This should give a warm chamber and even has a glass lid.
A practical question about your oven: These have a timer - do you have to activate it every 60 minutes or does it heat continuously?
That double temperature shut-off is a very good solution!
Bob
Re: Botanic section from paraffin block with unreadable label
Hi Bob, it has both options, a timer and a 'stay-on' option. I leave this on overnight set at 60 deg, no problems at all.
John B
Re: Botanic section from paraffin block with unreadable label
Ha Bob,
I went to a slide making training session last month, no practice in the wax-cutting area myself: they recommended adding 5% bee-wax to the mix to improve the cutting capability. An option when re-embedding?
I went to a slide making training session last month, no practice in the wax-cutting area myself: they recommended adding 5% bee-wax to the mix to improve the cutting capability. An option when re-embedding?
Re: Botanic section from paraffin block with unreadable label
Hi Maarten,
to which paraffin type did they add the bees wax? I would expect a commercial product to be optimized for the intended purpose. Botanic sections may be a bit outside the intended purpose.
The paraffin of these blocks cuts very bad, I don't think that it is worthwhile to try to improve it when something better is available. I have about two kilgramms of Paraplas Plus here and will cast the first blocks with it today. For the melting I use a cast aluminium cooking pot, with added silicone bottom and Armaflex heat insulation outside. This seems to work well - when I have used this a bit more and am still satified with the solution I can show it here.
Who offered the course you went to and how did you like it? Are you already preparing for paraffin embedding?
Bob
to which paraffin type did they add the bees wax? I would expect a commercial product to be optimized for the intended purpose. Botanic sections may be a bit outside the intended purpose.
The paraffin of these blocks cuts very bad, I don't think that it is worthwhile to try to improve it when something better is available. I have about two kilgramms of Paraplas Plus here and will cast the first blocks with it today. For the melting I use a cast aluminium cooking pot, with added silicone bottom and Armaflex heat insulation outside. This seems to work well - when I have used this a bit more and am still satified with the solution I can show it here.
Who offered the course you went to and how did you like it? Are you already preparing for paraffin embedding?
Bob
Re: Botanic section from paraffin block with unreadable label
I'd agree with you 100% there Bob - additions/modifications to Paraplast would surely be at best needless.
John B
Re: Botanic section from paraffin block with unreadable label
Bob,
Sorry, do not know what paraffin make they were referring to, no practice myself yet.
Still exploring hand cuts using a simple table microtome here.
Best,
DeBult
Sorry, do not know what paraffin make they were referring to, no practice myself yet.
Still exploring hand cuts using a simple table microtome here.
Best,
DeBult
Re: Botanic section from paraffin block with unreadable label
Hi together,
I have heard before that people are mixing bees wax under modern paraffin to make it more suitable for their use. Perfect sectioning depends probably on the combination of specimen, paraffin, knife and handling so I can very well imagine that people end up with different processes. When sommebody regularly uses a very sharp typ a or b knife he might have a different experience than somebody who uses a teflon coated single use blade with its comparatively blunt 35° wedge angle. Apart from that it is only half the fun if there is no room for magical recipes any more!
It would be interesting to speak to an expert old-timer who was used to source paraffin from where he was able to get it. How christmas tree candles feel like that are good to section...
Bob
I have heard before that people are mixing bees wax under modern paraffin to make it more suitable for their use. Perfect sectioning depends probably on the combination of specimen, paraffin, knife and handling so I can very well imagine that people end up with different processes. When sommebody regularly uses a very sharp typ a or b knife he might have a different experience than somebody who uses a teflon coated single use blade with its comparatively blunt 35° wedge angle. Apart from that it is only half the fun if there is no room for magical recipes any more!
It would be interesting to speak to an expert old-timer who was used to source paraffin from where he was able to get it. How christmas tree candles feel like that are good to section...
Bob