Abstract
Paraffin histology is one of the most important and commonly-used laboratory techniques in diagnostic histopathology. The discovery of paraffin embedding is often attributed to the pathologist Edwin Klebs. Klebs was following the lead of Stricker, who embedded embryos in a mixture of hot stearin and white beeswax. We show that Klebs experimented with paraffin wax for embedding tumour tissue. But he quickly rejected it as unsuitable because paraffin wax did not infiltrate the tissue. One of Klebs’ correspondents, embryologist Wilhelm His, Sr., learned of Klebs’ experiments and decided to try paraffin embedding. His dehydrated chicken embryos in alcohol, cleared them in lavender oil, and dripped hot paraffin wax onto them. This process allowed His to cut good sections. Here, we have replicated His’s paraffin embedding protocol in order to determine whether His had indeed made the landmark discovery of infiltration embedding with paraffin wax. We followed the protocol that he gives in his 1868 monograph on the early development of the chicken. The protocol described by His failed, in our hands, to yield sections of the quality that he illustrates in his monograph. Typically, the tissue disintegrated when sectioned due to poor infiltration of the wax. Usable sections could only be obtained if His’s protocol was modified by melting the embedded embryos in fresh paraffin wax. One explanation for our findings is that we failed to faithfully replicate His’s protocol. Another is that his protocol was incomplete. We suggest that His is likely to have discovered and perfected infiltration embedding with paraffin wax but did not publish a complete protocol.Supplementary InformationThe online version of this article (10.1007/s00292-021-00947-4) contains supplementary material, which is available to authorized users.
Highlights
Embedding is a technique used to prepare tissues for microscopic analysis
When molten paraffin was dripped onto embryos with approximately 20 μL lavender oil adhering, the paraffin failed to penetrate the tissue
We speculated that the paraffin was not molten long enough for it to mix with the lavender oil and infiltrate the tissue
Summary
Embedding is a technique used to prepare tissues for microscopic analysis. It entails the placing of the specimen in a solid mass while it is sectioned using a microtome [22]. There are two types of embedding [7]: peripheral embedding encases the tissue, supporting it only on the outside. Infiltration or interstitial embedding supports the tissue outside and inside, because the embedding mass completely permeates the tissue. Infiltration may require an intermediate reagent: a solvent that is miscible both with the alcohol used to dehydrate the tissue, and with the embedding medium [7, 21, 22]. Many intermediate reagents act as clearing reagents, rendering the tissue optically transparent [22, p. 68]
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