Skip to main content
SpringerLink
Log in

Principles of In Vivo Confocal Microscopy

  • Chapter
  • First Online:
In Vivo Confocal Microscopy in Eye Disease

  • 370 Accesses

Abstract

Three-dimensional visualization of biological tissues has been a challenge over the years. A number of modern digital optical technologies have been developed and in vivo confocal microscopy (IVCM) is one of the optical imaging techniques that has been widely used in research and, more recently, in specialist ophthalmology clinics.

The principle of IVCM is based on confocality of the light source and the detector plane. The type of confocal microscope design can be determined by its method of scanning and how the images are constructed. Modern confocal microscopes have better image quality due to new scanning technologies that decrease image artifacts. In addition, non-contact in vivo confocal microscopy has also been developed in which the contact cap over the microscope is removed and replaced by long focal length objective lenses.

In this chapter, the principles of IVCM are reviewed and the various types of confocal microscopes will be discussed including:

  • Laser scanning confocal microscope (LSCM)

  • Tandem scanning confocal microscope (TSCM)

  • Slit-scanning confocal microscope

  • Swept-field confocal microscopy (SFC)

  • Noncontact confocal laser scanning microscopy

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
eBook
USD 16.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 99.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Jalbert I, Stapleton F, Papas E, Sweeney DF, Coroneo M. In vivo confocal microscopy of the human cornea. Br J Ophthalmol. 2003;87(2):225–36.

    Article  CAS  Google Scholar 

  2. Wells WA, Thrall M, Sorokina A, Fine J, Krishnamurthy S, Haroon A, Rao B, Shevchuk MM, Wolfsen HC, Tearney GJ, Hariri LP. In vivo and ex vivo microscopy: moving toward the integration of optical imaging technologies into pathology practice. Arch Pathol Lab Med. 2019;143(3):288–98.

    Article  Google Scholar 

  3. Elliott AD. Confocal microscopy: principles and modern practices. Curr Protoc Cytom. 2020;92(1):e68.

    PubMed  PubMed Central  Google Scholar 

  4. Minsky M. Memoir on inventing the confocal scanning microscope. Scanning. 1988;10(4):128–38.

    Article  Google Scholar 

  5. Patel DV, McGhee CN. Contemporary in vivo confocal microscopy of the living human cornea using white light and laser scanning techniques: a major review. Clin Exp Ophthalmol. 2007;35(1):71–88.

    Article  Google Scholar 

  6. Girkin JM, Ferguson AI, Wokosin DL, Gurney AM. Confocal microscopy using an InGaN violet laser diode at 406nm. Opt Express. 2000;7(10):336–41.

    Article  CAS  Google Scholar 

  7. Guthoff RF, Zhivov A, Stachs O. In vivo confocal microscopy, an inner vision of the cornea–a major review. Clin Exp Ophthalmol. 2009;37(1):100–17.

    Article  Google Scholar 

  8. Paddock SW. Principles and practices of laser scanning confocal microscopy. Mol Biotechnol. 2000;16(2):127–49.

    Article  CAS  Google Scholar 

  9. De Silva ME, Zhang AC, Karahalios A, Chinnery HR, Downie LE. Laser scanning in vivo confocal microscopy (IVCM) for evaluating human corneal sub-basal nerve plexus parameters: protocol for a systematic review. BMJ Open. 2017;7(11):e018646.

    Article  Google Scholar 

  10. Patel DV, Zhang J, McGhee CN. In vivo confocal microscopy of the inflamed anterior segment: a review of clinical and research applications. Clin Exp Ophthalmol. 2019;47(3):334–45.

    Article  Google Scholar 

  11. Guthoff RF, Baudouin C, Stave J. Atlas of confocal laser scanning in-vivo microscopy in ophthalmology. Springer Science & Business Media; 2007.

    Google Scholar 

  12. Prydal JI, Dily PN. Advances in confocal microscopy of the cornea. Eye. 1998;12(3):331–2.

    Article  Google Scholar 

  13. Petráň M, Hadravský M, Egger MD, Galambos RO. Tandem-scanning reflected-light microscope. JOSA. 1968;58(5):661–4.

    Article  Google Scholar 

  14. Stachs O, Guthoff RF, Aumann S. In vivo confocal scanning laser microscopy. High Resolu Imag Microsc Ophthalmol. 2019:263–84.

    Google Scholar 

  15. Toomre DK, Langhorst MF, Davidson MW. Introduction to spinning disk confocal microscopy. ZEISS Campus. http://zeisscampus.magnet.fsu.edu/articles/spinningdisk/introduction.html. Accessed 2019. 2012.

  16. Bayguinov PO, Oakley DM, Shih CC, Geanon DJ, Joens MS, Fitzpatrick JA. Modern laser scanning confocal microscopy. Curr Protoc Cytom. 2018;85(1):e39.

    Article  Google Scholar 

  17. Wiegand W, Thaer AA, Kroll P, Geyer OC, Garcia AJ. Optical sectioning of the cornea with a new confocal in vivo slit-scanning videomicroscope. Ophthalmology. 1995;102(4):568–75.

    Article  CAS  Google Scholar 

  18. Masters BR, Thaer AA. Real-time scanning slit confocal microscopy of the in vivohuman cornea. Appl Opt. 1994;33(4):695–701.

    Article  CAS  Google Scholar 

  19. Castellano-Muñoz M, Peng AW, Salles FT, Ricci AJ. Swept field laser confocal microscopy for enhanced spatial and temporal resolution in live-cell imaging. Microsc Microanal. 2012;18(4):753–60.

    Article  Google Scholar 

  20. Webb RH, Hughes GW, Delori FC. Confocal scanning laser ophthalmoscope. Appl Opt. 1987;26(8):1492–9.

    Article  CAS  Google Scholar 

  21. Zhivov A, Stachs O, Stave J, Guthoff RF. In vivo three-dimensional confocal laser scanning microscopy of corneal surface and epithelium. Br J Ophthalmol. 2009;93(5):667–72.

    Article  CAS  Google Scholar 

  22. Stave J, Guthoff R. Darstellung von Tränenfilm und In-vivo-Kornea: Erste Untersuchungsergebnisse mit einem modifizierten konfokalen Laser-Scanning-Ophthalmoskop. Ophthalmologe. 1998;95(2):104–9.

    Article  CAS  Google Scholar 

  23. Pritchard N, Edwards K, Efron N. Non-contact laser-scanning confocal microscopy of the human cornea in vivo. Cont Lens Anterior Eye. 2014;37(1):44–8.

    Article  Google Scholar 

Download references

Acknowledgement

We wish to express our great appreciation to Maryam Kasiri for her assistance in writing this chapter.

Disclosures

None to declare.

Author information

Authors and Affiliations

  1. Department of Cornea and External Diseases, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran

    Parisa Abdi & Mehrnaz Atighehchian

Authors
  1. Parisa Abdi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mehrnaz Atighehchian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Parisa Abdi .

Rights and permissions

Reprints and permissions

Copyright information

© 2022 Springer-Verlag London Ltd., part of Springer Nature

About this chapter

Check for updates. Verify currency and authenticity via CrossMark

Cite this chapter

Abdi, P., Atighehchian, M. (2022). Principles of In Vivo Confocal Microscopy. In: In Vivo Confocal Microscopy in Eye Disease. Springer, London. https://doi.org/10.1007/978-1-4471-7517-9_1

Download citation

  • DOI: https://doi.org/10.1007/978-1-4471-7517-9_1

  • Published:

  • Publisher Name: Springer, London

  • Print ISBN: 978-1-4471-7516-2

  • Online ISBN: 978-1-4471-7517-9

  • eBook Packages: MedicineMedicine (R0)

Publish with us

Policies and ethics

Search

Navigation