I am looking for a protocol/procedure to obtain ultra thin histological sections that doesn't involve the use of rotary microtome. Please let me know about any other simple (easy-to-follow) and effective technique(s) or tool(s)/equipment(s) that can be used as an alternative yet better approach.
Use a cubical piece of peeled potato- Cut vertically from top to half length and place the washed plant part by slight widening the slit. Hold tight and use sharp razor or blade in a way to slide repeatedly through the tissue and keep quickly in clean water-select and stain either temp. or permanant.
Dr. Faiq, thank you for the method you mentioned. That's a quite conventional yet effective method.
Some other innovative approaches are also welcome.
The five main stages in the preparation of histology slides are:
Fixing
Clearing and Fixation
Embedding
Block making, Trimming the block, Mounting the block
Section cutting
Mounting and spreading the ribbon
Labeling
Staining Process
Just to add to the previous answer: instead of a potato you can also use carrots or any other similar plant tissue and make sure you are using a sharp razor or interchangeable scalpels. Also, chances are that only 1 in 20 sections will be successful as the method requires good manual precision.
“Ultra thin sections” are usually for transmission electron microscopy (TEM) and the thickeness is around 0.1 micro meter. It is impossible to cut without microtome.
Even in the case of sections for biological optical microscopes, resin-embedding and thin sections around 1-10 micron meters thick cut using a rotary microtome equipped with a glass knife is recommended to obtain superior images.
One of the conventional techniques to cut plant material such as a leaf, a pith of Sambucus spp. is used ad Dr. Faiq described. In these days, styrene foam popularly used for packaging can be used in stead of the pith.
Sharp blades are available from a disposable razor, which you may use daily.
The trick is to select finely cut parts.
The question is clear that the requirement is to obtain a plant material section without microtome (No Rotatory, no sliding). The only option will be hand processed section using a razor and pith to support plant material with fine sliding the razor repeatedly.
Can you understand my suggestion on August 22?
When asking something for a person, you should clarify what you are seeking. This time, I can easily understand that you did not need "ultrathin sections". You should clarify what kind of microscopic images you are seeking and explain your situation without a rotary microtome or a sliding microtome. So that I recommend you a traditional method well-known in plant anatomy.
On the other hand, you should have looked for friends or organizations that could let you use the necessary equipment nearby. It is common sense in a Japanese research society.
Dear Dr. Tomoyuki, thank you for your extended explanation. It was quite clear from your first answer that there is NO WAY to obtain ultra-thin section without TEM or to obtain semi-thin section without microtome. My question was on probable alternative options to get such sections if one doesn't have such instrumentation facilities any way. Eventually, it stands that the only option left is the conventional "thin" sectioning using disposable sharp razors and styrene foam or pith. Your comprehensive statement is appreciable.
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