Tooth formation in a five-day-old mouse in different histological staining.
The two yellow circles in the image indicate the location of tooth development. On the left the mandible and on the right the maxilla of a five-day-old mouse. Fixed in Bouin and stained with PTAH according to Mallory.
Teeth develop through the interaction between the ectoderm[1] of the oral cavity and the special mesenchyme[1] underneath, which originates from the neural crest and is called ectomesenchyme.
A tooth bud differentiates through the tooth cap stage to the tooth bell with outer and inner enamel epithelium.
All images shown here are of the bell stage.
The odontoblasts form the non-calcified predentin from which the calcified dentin[1] is formed.
Dentin can be formed by the odontoblasts throughout life and, like bone, consists of organic material, mainly type 1 collagen and 70% inorganic hydroxyapatite[1].
The ameloblasts form the enamel prism. Ameloblasts are long narrow cells with an elongated light-colored nucleus.
Enamel is formed only during tooth development by the ameloblasts of the inner enamel epithelium and consists of about 95% inorganic constituents, mainly hydroxyapatite. The enamel forms the outer layer of the dental crown[2].
All
images shown here were taken on a Leitz Orthoplan microscope with Fluotar and
Apo objectives and a Moticam 2300 camera.
1 = enamel pulp;
2 = stratum
intermedium;
3 = ameloblasts
(inner enamel epithelium);
4 = secretory
granulum;
5 = enamel;
6 = dentin;
7 = predentin;
8 = odontoblasts;
9 = dental papilla (early
dental pulp).
Stain-protocol of the applied PTAH staining. [3]
PTAH (Fosforwolfraamheamatoxyline) [3] | |
Bring paraffin coups via xylol and descending alcohol series in water | steps of 4 min |
| Oxidise in kaliumpermanganaat 0,25% | 10 min |
Rinse AD | |
Oxalic acid 5% (removal of mangaandioxide) | 10 min |
| Rinse AD | |
| Fosforwolfraamhaematoxyline | 16 hrs |
Ethanol 96% (do not rinse with AD first) | ≈ 1 min |
Ethanol 96% | 1 min |
| Isopropanol 100% 2x | 4 min |
| Xylol 1 | 4 min |
| Xylol 2 | 4 min |
| Embed in example Depex, Euparal, Malinol | |
Fosforwolfraamhaematoxyline: - A: 1g haematoxyline, solve under heating in 50ml AD; - B: 20g phosphorus tungstic acid solve in 50ml AD; - Mix A and B and allow to mature for several weeks (possibly artificially mature by adding 0.177g of potassium permanganate). (It is convenient to ripen in, for example, an open conical flask and shake occasionally. Cover with an inverted coffee filter, for example). |
Source:
[2] Prof. Dr. med. Dr. rer. nat. Ulrich Welsch, Elsevier GmBH, Urban&Fischer Verlag München, Sobotta Welsch Lehrbuch Histologie (2006, tweede druk), Hoofdstuk 10, pag. 342 - 346, 'Verdauungsorgane', ISBN 978-3-437-44430-2.
[3] Romeis, Prof. Dr. Peter Böck, Urban & Schwarzenberg München 1989, Mikroskopische technik (1989, zeventiende druk), Hoofdstuk 25, pag. 507, 'Binde- und Stützgewebe', par. 1.3.4.4, Phosphorwolframhämatoxylin nach Mallory, ISBN 3-541-11227-1
[4] Romeis, Prof. Dr. Peter Böck, Urban & Schwarzenberg München 1989, Mikroskopische technik (1989, zeventiende druk), Hoofdstuk 8, pag. 235, 'Färben', par. 2.5.1, Doppelfärbung mit Hämalaun-Eosin (H&E), ISBN 3-541-11227-1
[5] Romeis, Prof. Dr. Peter Böck, Urban & Schwarzenberg München 1989, Mikroskopische technik (1989, zeventiende druk), Hoofdstuk 25, pag. 499, 'Binde- und Stützgewebe', par. 1.3.1.4, Trichromfärbung nach Goldner, ISBN 3-541-11227-1
[6] Romeis, Prof. Dr. Peter Böck, Urban & Schwarzenberg München 1989, Mikroskopische technik (1989, zeventiende druk), Hoofdstuk 25, pag. 501, 'Binde- und Stützgewebe', par. 1.3.1.7, Azanfärbung nach Heidenhain, ISBN 3-541-11227-1
[7] Romeis, Prof. Dr. Peter Böck, Urban & Schwarzenberg München 1989, Mikroskopische technik (1989, zeventiende druk), Hoofdstuk 8, pag. 236, 'Färben', par. 2.5.2, Dreifachfärbung nach van Gieson (1889), ISBN 3-541-11227-1