Confocal microscopy is a noninvasive imaging technique that enables high-resolution analysis of ocular surface microstructure. It was invented in 1957 and works by illuminating samples point-by-point and rejecting out-of-focus light to generate optical sections. There are various types including tandem scanning, scanning slit, and laser scanning confocal microscopes. Clinical applications include imaging corneal layers, assessing wound healing after refractive surgery, and diagnosing infections. It allows evaluation of conditions like diabetic keratopathy by quantifying changes in subbasal nerve fibers. Advanced techniques like second harmonic generation further study corneal collagen architecture.

1. CORNEAL CONFOCAL MICROSCOPY
DIYAR J.K.

2. CONFOCAL MICROSCOPY
• is an emerging noninvasive imaging and diagnostic tool.
• enables morphological and quantitative analysis of
ocular surface microstructure, both in health and
disease.
• The principal of confocal microscopy was patented in
1957 by Marvin Minsky while he was a junior fellow at
Harvard University.
• The key elements of Minsky's confocal microscope
design: the pinhole apertures, point-by- point illumination
of the specimen and also rejection out-of-focus light

4. TRY TO THINK

5.  (TSCM) which used a spinning disk containing
thousands of optically conjugate (source/detector)
pinholes arranged in Archimedean spirals
TYPES : Tandem scanning confocal microscope

6. TRIAL TO THINK OUT OF BOX

7. Scanning slit confocal microscopy
• The Confoscan 4 confocal microscope : uses a slit
scanning design has 20× lens that works 12 mm from
the corneal apex for endothelial cell count, allowing fully
no-contact exam with z-adapter
• provides images of all corneal cell layers, albeit with less
resolution than the HRT-RCM. Is particularly well suited
to imaging the corneal endothelial cells.
• Due to its thicker optical volume, full-field images are
easily obtained.the optical sectioning capability of
confocal microscopy allows imaging through edematous
corneas.

9. BE CREATIVE

10. Laser scanning confocal microscopy
The HRT III with Rostock Corneal Module (HRT-RCM).
has 63× lens and optional 10 × to see deeper (lens,zonules)
uses 670 nm diode laser and provides 1micron resolution.
HRT-RCM does applanate cornea and the use of PMMA
disposabel cap and gel is mandatory for the exam.]

13. TEAR FILM

14. EPITHELIUM

16. BOWMAN LAYER
• Changes in the pattern of innervation can be assessed
using quantitative outcome measures such as nerve
density, length, branching &tortuosity.

17. STROMA

18. ENDOTHELIUM

20. ONE OF THE THEORY OF CREATIVITY

21. Confocal microscopy through-focusing (CMTF)
• To collect and quantify 3-D information from the cornea
• CMTF scans are obtained by focusing through the
cornea from the epithelium to endothelium at a constant
lens speed, while continuously digitizing images.
• The CMTF intensity curve is generated by calculating the
average pixel intensity in a central region of each image
& plotting versusz depth.

24. • Because of its unique ability to image the cornea four-
dimensionally at the cellular level.
• allows measurement of wound gape, the depth of
epithelial ingrowth & the degree of corneal fibrosis in
radial keratotomy wounds.
• Confocal microscopy is especially well suited to
assessing the corneal response to PRK and
LASIK,LASEK & epiLASIK as well as other surgical
techniques such as DSAEK.
CLINICAL APPLICATION : Wound healing following
injury or refractive surgery

27. Infectious keratitis ..ACANTHAMOEBA K.

28. FUNGAL KERATITIS

29. Imaging changes in the sub basal nerve plexus
• Following PRK, regenerating sub-basal nerve fibers were
detected as early as7 days after PRK,but required at
least 1–2 years for complete recovery, perhaps
due to the persistence of the subepithelial scarring.
Following LASIK, sub-basal nerve density remained
lower for 3–5 years after surgery
• significant differences have been identified between
normal and diabetic subjects, which may precede the
development of diabetic retinopathy.

32. • To studying the structure of the cornea, since collagen
fibrils generate strong SHG signals that can be imaged
using optical microscopic techniques.
• To generate an SHG signal, a material is excited
by the simultaneous absorption of two photons of light of
the same frequency and then the material emits a single
photon of light that is double the frequency or half the
wavelength of the excitation light.
• The ability to generate SHG signals is limited to
materials that are highly ordered and non-
centrosymmetric, such as collagen.
Advanced Technology:Second Harmonic Generation
Imaging of Corneal Collagen Architecture

35. THANK YOU