1.The corneal universe - mission possible by in vivo confocal microscopy
Jonathan Kersley Lecture
Presented by
Christina Grupcheva

2. Jonathan Kersley
(1939-2000)

Henry Jonathan Kersley was born in Bath

3. Educated at Cambridge University 4. Completed medical studies at St Bartholomew's Hospital 5. Member of the Royal College of Ophthalmologysince founded in 1988 6. In 1974, he gave his first paper at the international meeting of theContact Lens Society in Montreux

He was involved in clinical development of both Sauflon 70 and

Sauflon85 by Contact Lenses Manufacturing Ltd

Before implants were universally considered, he was possibly the

first to fit soft lenses immediately after cataract surgery

Kersley was President of the British Contact Lens Association 1978-9

'Modern Anterior Segment Surgery and advancing contact lens technology'
7. Historical prospective

1957Minsky describes confocal microscope principle

8. 1980In vitro confocal microscopy well established 9. 1985Lemp et al apply in vivo technique to rabbit cornea 10. 1989Cavanaugh et al publish first human data 11. 1995 Slit-scanning in vivo confocal microscopy 12. 2003 Laser-scanning in vivo confocal microscopyJonathan Kersley Lecture 2011
13. Tandem-scanning in vivo confocal microscopy
White light
Scanning spot
or scanning slit
Incident light is
focussed in same
plane as objective
lens i.e. confocal
Objective
Nipkow
lens
Beam
Disk
Light
splitter
source
Video
camera
CORNEA
Dual light path technique (Tandem scanning)
Single light path technique
Jonathan Kersley Lecture 2011
14. Slit-scanning in vivo confocal microscopy
Light
source
Condenser
lens
Front
lens
Scanning
slits
Condenser
lens
Video
camera
Cornea
Jonathan Kersley Lecture 2011
15. Confoscan 2 (model 2000)
improved NAVIS software
Confoscan 2 (model 1999)
In vivo confocal microscopy
Jonathan Kersley Lecture 2011
16. Tandem scanning confocal microscopy
Precise movement of the disk
Applanation technology ?
Small round area
Low illumination
One scan per examination
Time consuming
Slit- scanning confocal
microscopy
Greater illumination
Adjustable parameters
Relatively quick
Non-applanation technology
Small oblong area
Moving objective lens
In vivo confocal microscopy
Jonathan Kersley Lecture 2011
17. Laser-scanning in vivo confocal microscopy
Jonathan Kersley Lecture 2011
18. In vivo confocal microscopy
Light-scanning confocal microscopy
Non applanation
Clearer images of the endothelium
Depends on transparency
Low resolution
Artefacts
Time consuming
Laser- scanning confocal
microscopy
Greater resolution
Greater depth of focus
More compact, stabile objective
Precise z location ?
Small area
Low reproducibility
Jonathan Kersley Lecture 2011
19. Research & clinical applications
Clinical:
Description
Differentiation
Diagnosis
Prognosis
Dynamic follow up
Research:
Descriptor
Discriminator
Dissector
3D reconstructor
4D reconstructor
20. 2
3
1
1
2
3
4
5
4
5
6
6
7
8
7
8
Research applications:
Descriptor & Discriminator
Epithelium
Anterior stroma
Posterior stroma
Endothelium
21. Research applications:
Descriptor & Discriminator
2
3
1
1
Epithelium
2
3
Anterior stroma
4
5
4
5
6
6
Posterior stroma
7
8
7
Endothelium
8
22. Research applications:
Descriptor & Discriminator
Epithelium
Anterior stroma
Posterior stroma
Endothelium
23. Quantitative
analysis
Research applications:
Descriptor & Discriminator
Basal epithelium
Qualitative analysis:
Honey-comb appearance
Bright borders
Dark homogenous bodies
Jonathan Kersley Lecture 2011
24. Quantitative
analysis
Research applications:
Descriptor & Discriminator
Wing epithelial cells
Qualitative analysis:
50% presence
resembling fried eggs
bright borders
homogenous cytoplasm
bright nuclei
Occasionally:
very bright bodies
grouped in duplets,
bigger groups
up to 10 cells
Jonathan Kersley Lecture 2011
25. Research applications:
Descriptor & Discriminator
Sub-basal nerve plexus
Qualitative analysis:
Bright, linear
Branching
Beadings
Jonathan Kersley Lecture 2011
26. ...thickness of the sample...
5 mm
3 mm
however, confocal microscopy is not perfect...
...optical shadowing...
ghost images...
...artefacts...
?
27. Research applications:
Descriptor & Discriminator
AnalySIS
Nerve density:
- calibration
- outlining
- reading
Jonathan Kersley Lecture 2011
28. Research applications:
Descriptor & Discriminator
AnalySIS
Nerve diameter:
10 randomly
distributed
readings/slide
Jonathan Kersley Lecture 2011
29. Research applications:
Descriptor & Discriminator
AnalySIS
Nerve beadings:
N of beadings
per 100 mm
Jonathan Kersley Lecture 2011
30. Research applications:
Descriptor & Discriminator
Group 1Group 2Statistical
significance
Nerve 632.35287.57 582.39327.13 P