Introduction to confocal microscopy

Adaobi Nwaneshiudu, Christiane Kuschal, Fernanda H. Sakamoto, R. Rox Anderson, Kathryn Schwarzenberger, Roger C. Young

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Confocal microscopy is a technique in optical imaging that uses point illumination via a spatial pinhole to eliminate outof-focus signals. The excitation light in confocal microscopy is usually provided by a laser to generate high intensities of fluorescence or reflectance from the focal spot. Fluorescence confocal microscopy is the most used in dermatology to analyze ex vivo and in vitro samples. Reflectance confocal microscopy can be used for real-time microscopy and uses melanin as a natural contrast agent. Confocal microscopy has many advantages, including increasing the optical resolution and contrast of an image of a specimen; facilitating reconstruction of 3-D images; enabling collection of serial optical sections from thick specimens; and enabling in vivo imaging without the artifact induced by tissue processing (Pawley, 2006). In addition to LSCM, 3-D images of nonliving samples can also be acquired by SCEM, where an electron beam is used for illumination, resulting in higher resolution compared with onfocal microscopy. Limitations of confocal microscopy include the depth of imaging within thick samples and cost compared with conventional microscopes. The problems of fluorescent probe photobleaching and phototoxicity inherent in conventional fluorescence microscopy are also present with confocal microscopy. Multiphoton microscopy is an alternative strategy for fluorescence microscopy, which offers higher resolution, somewhat greater depth of imaging, and minimal photobleaching. Technologies for microscopy are promising and are still being improved.

Original languageEnglish (US)
Pages (from-to)1-5
Number of pages5
JournalJournal of Investigative Dermatology
Volume132
Issue number12
DOIs
StatePublished - Dec 1 2012
Externally publishedYes

Fingerprint

Confocal microscopy
Confocal Microscopy
Microscopy
Fluorescence microscopy
Microscopic examination
Fluorescence Microscopy
Photobleaching
Imaging techniques
Three-Dimensional Imaging
Lighting
Dermatology
Phototoxic Dermatitis
Optical Imaging
Melanins
Fluorescent Dyes
Artifacts
Contrast Media
Electron beams
Lasers
Microscopes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology
  • Cell Biology

Cite this

Nwaneshiudu, A., Kuschal, C., Sakamoto, F. H., Rox Anderson, R., Schwarzenberger, K., & Young, R. C. (2012). Introduction to confocal microscopy. Journal of Investigative Dermatology, 132(12), 1-5. https://doi.org/10.1038/jid.2012.429

Introduction to confocal microscopy. / Nwaneshiudu, Adaobi; Kuschal, Christiane; Sakamoto, Fernanda H.; Rox Anderson, R.; Schwarzenberger, Kathryn; Young, Roger C.

In: Journal of Investigative Dermatology, Vol. 132, No. 12, 01.12.2012, p. 1-5.

Research output: Contribution to journalArticle

Nwaneshiudu, A, Kuschal, C, Sakamoto, FH, Rox Anderson, R, Schwarzenberger, K & Young, RC 2012, 'Introduction to confocal microscopy', Journal of Investigative Dermatology, vol. 132, no. 12, pp. 1-5. https://doi.org/10.1038/jid.2012.429
Nwaneshiudu A, Kuschal C, Sakamoto FH, Rox Anderson R, Schwarzenberger K, Young RC. Introduction to confocal microscopy. Journal of Investigative Dermatology. 2012 Dec 1;132(12):1-5. https://doi.org/10.1038/jid.2012.429
Nwaneshiudu, Adaobi ; Kuschal, Christiane ; Sakamoto, Fernanda H. ; Rox Anderson, R. ; Schwarzenberger, Kathryn ; Young, Roger C. / Introduction to confocal microscopy. In: Journal of Investigative Dermatology. 2012 ; Vol. 132, No. 12. pp. 1-5.
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