Hello,
To compare root canal irrigation techniques I am currently doing an experiment where I grow bacteria inside a tooth and then clean it using different irrigation. I then cut it into 500microns slices with a diamond wire saw, stain it with BacLight live/dead stain and image it with a confocal microscope (CLSM).
The problems are :
- The diamond wire saw cannot make thin slices thus the CLSM cannot image through all the sample.
- The debris created by the wire saw contaminate the samples.
Do you have a suggestion on a method of histology I could use to cut the tooth into
Hello
What might possibly work is a saw microtome (Innenlochsaege).
e.g. Leica SP1600 saw microtome
It can cut unimbedded hard materials very thin.
see:
https://vimeo.com/136895449
What you cannot avoid is debris contaminating neighoring soft structures like your bacteria and I supose you would have to cut without water cooling.
It seems that Prof. Mathias Zehnder from Department of Preventive Dentistry, Periodontology, and Cariology, University of Zürich
Center for Dental Medicine had access to such an instrument at one time. Maybe it is still there.
Nice question.
I recently chanced a paper which I believe could provide an answer to your question. Article A new improved protocol for in vitro intratubular dentinal b...
Another one in nature publications
Confocal imaging identified the epoxygenase 2C23 isoform as the responsible isoform for 11,12-EET synthesis that maintains normal blood pressure in the face of salt loading (74).
Kindly find the full article here https://www.sciencedirect.com/topics/engineering/confocal-imaging
Dear Clement,
in our lab, we use a variant of the classical embedding technique to get tooth sections thinner than 50 µm. The epoxy embedded tooth is sectioned in the desired plane with a water cooled, diamond coated cutting wheel. The sectioned surface is polished and mounted to a glas slide. After that, the block is sectioned from the other side and the remaining slab on the glas slide is ground and polished to the final thickness. The exposed section surface could be treated with dyes but probably the whole procedure has the potential to contaminate your samples.
An alternative would be using a laser microtome. A former colleague is now working with a company that build such devices and are also offering customized cuts (https://www.lls-rowiak.de/). The laser pulses turn material into plasma and you are able to create thin slices or blocks. This might be a way of non-contact testing.
Hope this might help
Carsten
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining