Traditional Microscopic Techniques Employed in Dental Adhesion Research—Applications and Protocols of Specimen Preparation
Abstract
:1. Introduction
2. Search Strategy
3. Confocal Laser Scanning Microscopy (CLSM)
3.1. Historical Background
3.2. Principles of Technique
3.3. Specimen Preparation
3.4. CLSM in Dental Adhesion Research
3.5. Limitations
3.6. Future Perspectives
4. Scanning Electron Microscopy (SEM)
4.1. Historical Background
4.2. Principle of the Technique
4.3. Specimen Preparation
4.4. SEM in Dental Adhesion Research
4.5. Limitations
5. Transmission Electron Microscopy (TEM)
5.1. Historical Background
5.2. Principle of Technique
5.3. Specimen Preparation
5.4. TEM in Dental Adhesion Research
5.5. Limitations
5.6. Future Perspectives of SEM and TEM
6. Atomic Force Microscopy (AFM)
6.1. Historical Background
6.2. Principle of the Technique
6.3. Specimen Preparation
6.4. AFM in Dental Adhesion Research
6.5. Limitations
6.6. Future Perspectives
7. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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Technique | Optical Microscopy | Electron Microscopy | Scanning Probe Microscopy | |||
---|---|---|---|---|---|---|
Conventional | Fluorescent | Confocal Scanning Microscope | Transmission Electron Microscope | Scanning Electron Microscope | Atomic Force Microscope | |
Source of energy | light (380–750 nm) | Specific light wavelength (absorbed by the fluorophores in the specimen) | Localized laser excitation and fluorescence effect | The interaction of beam of electrons that is transmitted through the specimen | Beam of high-energy electrons that interact with the specimen surface and generate a variety of signals | Deflection of the tip caused by atomic and electrostatic forces between the tip and specimen |
Magnification | To 100–1000× | 2,000,000–50,000,000× | To 100,000,000× | |||
Resolution | 100–1000 nm | 1–10 nm | 0.2 nm (horizontal) 0.05 nm (vertical) | |||
Main implementations |
|
|
|
Search Strategy | Inclusion Criteria | Exclusion Criteria |
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Type of paper | Systematic reviews, metanalyses, original research papers published | - |
Form of publication | Full-text or book chapter | Abstract only |
Language of publication | English | Non-English publications |
Publication content | Relevant to the subject of dentistry and dental research | Not including the content associated with dental tissues or dental materials |
Procedure | CLSM | SEM, TEM | AFM |
---|---|---|---|
Sectioning | Water-cooled diamond wafering blade | ||
Cleaning and polishing |
| ||
Specific recommendations according to the method | Labelling with fluorescent dye (i.e., rhodamine B) | Dehydratation and fixation Covering with electro-conducting coating layer | Flat specimen surface is necessary (Specimen labelling, dehydration, fixation and covering is not required) |
Specimen mounting | To the glass slide with cyanoacrylate glue | To the aluminum stub with double-sided carbon or copper tape | With a double-sided tape/glue to the steel disk that is finally mounted into magnetic specimen |
Area of Dentistry | Number of Items in PubMed Database (Year 2020) | Subject of SEM Research |
---|---|---|
Restorative dentistry, prosthetics and dental materials | 206 |
|
Biomaterials and guided tissue regeneration | 26 |
|
Implantology | 56 |
|
Orthodontics | 12 |
|
CLSM [4,11,12,17,78] | SEM [17,30,69,78] | TEM [39,41,42] | AFM [3,7,66,69,75] | |
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Advantages |
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Disadvantages |
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Author, Year | Research Topic | Microscopic Techniques | Observed Parameters | Authors’ Conclusions |
---|---|---|---|---|
Wu, et al. (2020) [81] | Remineralization of caries lesions in dentin after application of bioactive glass (BAG) | AFM, CLSM | AFM—surface topography, microhardness, the depth of the remineralization CLSM—fluorescence on the superficial layer of the lesion (decrease in fluorescence correlated with remineralization) | BAG obtains a promising remineralization effect |
Olley, et al. (2020) [82] | Dentin tubule patency and surface roughness after novel dab-on or brushing abrasion | CLSM, AFM, SEM, EDX, contact profilometry (CP) | CP—dentinal surface roughness CLSM—comparison of tubular patiency, AFM-roughness in intertubular region, SEM –the penetration of dentifrice EDX—constituents of deposits existing in dentinal tubules | Dab-on applications of either SnF2 or NaF dentifrice reduce the patency of dentine tubules and therefore reduce dentinal hypersensitivity |
Šugár et al. (2020) [83] | Laser machining of Ti-graphite composite for dental application | CLSM, SEM, EDX | CLSM, SEM—visualization of laser-prepared surface, EDX—the elemental composition | The thermal energy from laser put to the Ti-graphite composite has a positive effect on surface properties of dental implant |
Bastos-Bitencourt, et al. (2021) [16] | The effect of bioactive scaffolds of nanofibers on dentin remineralization | CLSM, SEM | CLSM—identification and comparison the dentinal tubule obliteration (magnification 40×) SEM—revealing the dentin morphology (magnification 3000×) | Desensitizing agents with nanofibers are potentially effective in dentin remineralization |
Pandele, et al. (2020) [84] | The method of synthesis new composite films based on polylactic acid and micro-structured hydroxyapatite particles (HA) | SEM (AFM) microscopy, FT-IR and, Raman spectroscopy, thermogravimetry (TG) | SEM, AFM—morphological analysis FT-IR and Raman spectroscopy- structural analysis TG—assessment of thermostability of the polymer. | The crystallinity of the composite films was decreased in comparison to the pure polymer. The presence of hydroxyapatite crystals did not have a significant influence on the degradation temperature of the composite film. |
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Nawrocka, A.; Piwonski, I.; Sauro, S.; Porcelli, A.; Hardan, L.; Lukomska-Szymanska, M. Traditional Microscopic Techniques Employed in Dental Adhesion Research—Applications and Protocols of Specimen Preparation. Biosensors 2021, 11, 408. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11110408
Nawrocka A, Piwonski I, Sauro S, Porcelli A, Hardan L, Lukomska-Szymanska M. Traditional Microscopic Techniques Employed in Dental Adhesion Research—Applications and Protocols of Specimen Preparation. Biosensors. 2021; 11(11):408. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11110408
Chicago/Turabian StyleNawrocka, Agnieszka, Ireneusz Piwonski, Salvatore Sauro, Annalisa Porcelli, Louis Hardan, and Monika Lukomska-Szymanska. 2021. "Traditional Microscopic Techniques Employed in Dental Adhesion Research—Applications and Protocols of Specimen Preparation" Biosensors 11, no. 11: 408. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11110408