Fluorescein

Chemical Properties:
1. C2 H12 O5 Na
2. Low molecular weight = 376.27 (can diffuse thru body fluids except retina and
choroidal vessels and diffuses hydrophilic device)
3. Orange-red in crystalline powder but yellow in solution
4. Fluorescence – influenced by:
a. yellow-green with cobalt-blue light in weak alkali solution
b. peak absorption: 465-490 nm
c. peak emission: 520-530 nm
d. pH – increase at pH 8 and loses at >8 (Quenching Phenomenon)
e. conc. – increase at 0.001% and loses at >0.001% called “Quenching Phenomenon,”
it dimerizes & polymerizes and emission shift to longer wavelengths

Clinical Characteristics:
1. Stains epithelial defects bright green by diffusing into intercellular spaces
2. Will not stain devitalized/dead cells or mucus
3. Tear film appears yellow-orange
4. Promotes growth of Pseudomonas aeruginosa
5. Will stain soft contact lens
6. Exhibit:
a. Pseudoflare – w/ epithelial breaks, fluorescein diffuses the corneal stroma and into
aqueous chamber producing greenish glow
KERATOCYTES
 b. Fisher-Schweizer’s corneal mosaic or Ant. corneal mosaic – fishnet-like mosaic pattern
of fluorescein in tear film after rubbing the eye w/c disappear if it is wash out
c. Negative staining

Indications: Schirmer’s test → Dry eyes → measures quantity of tears

1. Evaluate tear film layer (TBUT) → Dry eyes → measures quality of tear film
Causes of mucin deficiency:
a. vit. A deficiency
b. Stevens-Johnson syndrome → allergic reaction by meds.
c. ocular pemphigoid
2. Lacrimal drainage system (Jones test/Flourescein Dye Test)
Jones test I – can use dry cotton buds and placed at inferior turbinates of nose or by blowing
the nose with tissue to see the dye
Jones test II – irrigate lower canaliculus w/ NSS
results: a. blockage at duct – heavily stain buds after irrigation
b. blockage at canaliculi or at puncta – no fluorescein after irrigation
3. Aqueous wound leakage – after cataract surgery or penetrating injury
Seidel’s test – detect aqueous flow in conjunctival filtering blebs after trephine operation,
corneal laceration or post-cataract operation
*Fluorescein is instilled on eye and dye is seen flowing downward from surgical
or penetrating wound appearing as bright green in contrast to duller, yellow-orange
background.
4. Contact lens (RGP) fitting pattern
5. Retinal & choroidal vasculature (IV Fluorescein Angiography)
Layers of Pre-corneal Tear Film → evaporate water from the cornea → prevent
Dry eyes
1. Oily/Lipid layer → secreted by Meibomian/Tarsal glands
Function: prevent evaporation of water
2. Aqueous (water) layer → thickest layer
→ secreted by Lacrimal glands
Function: hydration
3. Mucin layer → secreted by Goblet cells (present at conjunctiva)
Functions:
a. wetting agent → enhance absorption
b. maintain integrity the whole layer tear film
Fisher-Schweizer’s Corneal Mosaic
TBUT
JONE’S TEST II
Jones Test II
Lacrimal Blockage
Lacrimal Probing
Seidel Test
6. Goldman applanation tonometry – define tear meniscus & precise applanation area
7. Identify presence of Atypical Mycobacterium, fungi, Herpes simplex I & Acanthamoeba infection
(protozoa)
8. Study of:
a. KCS – in relationship to accelerated epithelial loss
b. demonstrate human lacrimal ductules after dacryocystorhinostomy – Dye Disappearance
Test
*Fluorescein dye disappearance test (DDT) – reliability indices for anatomical and
subjective success after dacryocystorhinostomy
c. association of diabetic retinopathy w/ keratitis
S/E: transient eye irritation on cornea & conjunctiva

FLUOREXON
Chemical properties:
1. Carboxymethyl-aminoethyl-fluorescein tetrasodium
2. High molecular weight: 710 (prevent diffusing to contact lens & do not stain small epithelial
defects e.g. micropunctate keratitis, fine dendritic keratitis, small blebs,
minor corneal erosions
3. Stains large epithelial defects, devitalized cells (stains cytoplasm & nucleus) and mucin
4. Fluorescence
a. less brilliant
b. does not increase w/ increasing concentration
c. uses yellow filter
5. Does not stain most soft contact lenses
6. May stain high >60% water contents
7. Promotes growth of Pseudomonas aeruginosa
FLUORESCEIN ANGIOGRAPHY
Applanation Tonometer – Mires
DENDRITIC KERATIC PPT
 FLURESS
Contents:
1. 0.25% Na Fluorescein 4. 1% chlorobutanol - antiseptic
2. 0.4% Benoxinate HCl 5. 0.1% edetate disodium (chelating agent) and
preservative
3. 15% Povidone - antisepetic 6. Boric acid (buffering agent) to pH = 5
*<0.125% failed to stain the apex of meniscus Na Borate : alkaline
* >0.5% causes residual staining
S/E: Vasovagal responses in applanation tonometry causing syncope & grand mal seizure

1% - 2% ROSE BENGAL DYE (solution or strips)

Chemical characteristics:
1. Tetrachloro-tetraiodo fluorescein sodium
2. Stains devitalized cells and mucin Brilliant Red seen by naked eyes (does not enter epithelial
defects nor stain normal cells due to blocking effect of precorneal tear film)
3. Stains nucleus more than the cytoplasm
4. Fluorescence – increases w/ increasing concentration
5. Has antiviral property
6. Stains eyelids

Indications:
1. early diagnosis of KCS & Sjogren’s syndrome (tear deficiency)
2. characterizing the dendritic lesions of Herpes simplex & zoster lesions
Fluress
ROSE BENGAL DYE on Sclera and Cornea
 S/E:
1. transient stinging sensation – may add anesthetic but can damage/destroy cells
causing false (+) results for KCS
2. permanent corneal stromal deposits in large corneal or conjunctival epithelial defect
3. skin discoloration – more permanent
4. photothrombosis on neovascularization after exposure to Argon Laser or Irradiation
- thrombus remains for 16-18 weeks
*Rose Bengal should be used 1st before Fluorescein dye

1% INDOCYANINE GREEN (ICG)

Characteristics:
1. protein bound given IV angiography – pattern vessels, thrombus, hemorrhage
2. peak absorption: 805 nm
3. peak fluorescence: 835 nm in infrared spectrum
4. stains disease or dead endothelial cells - cornea
5. water-soluble tricarbocyanine
Indications:
1. determine cardiac output (CO)
2. determine liver outflow and functions
3. Angiography – more effective in revealing choroidal neovascular membranes due larger
wavelengths
S/E: rare (anaphylactic rxn due to iodofluorescein)
FA ANGIOGRAPHY ICG ANGIOGRAPHY
 1% ALCIAN BLUE
Corneal stromal dystrophy characterized by deposition of acid mucopolysaccharides in the corneal stroma
stain by alcian blue
Indication: differentiate mucin deposits (stained blue) from diseased cells (unstained)

10% - 20% ARGYROL

- silver protein with antiseptic property
- mucus strands & debris on lid margins & skin surface appear discolored but not normal tissue
Indication: excellent indicator of adequacy of preoperative preparation prior to surgical
eye scrub (absence of stain means preparation is complete)

SULFORHODAMINE B
Characteristics:
1. molecular weight: 559 (does not stain epithelial cells but stain corneal stroma)
2. less lipid soluble
3. Fluorescence
a. orange excited by green light (by viewing thru orange filter can eliminate
fluorescence of sclera allowing observation of tear film)
b. peak absorption: 556 nm
c. peak emission: 572 nm
S/E & C/I: none
ALCIAN BLUE at Corneal Stroma
Eye Scrub with Argyrol
Sulforhodamine B
 5% METHYLENE BLUE
Characteristics:
1. vital stain of corneal nerve tissue and stain devitalized cells & mucin
2. bluish ocular discoloration persisting for 1 day
3. has bacteriostatic property
4. precipitate in alkaline solution
5. peak absorption: 660 nm
Indication: for irrigation of lacrimal sac prior to Dacryocystorhinostomy
S/E: more eye irritation (thus combined w/ anesthetic)

1% LISSAMINE GREEN
Characteristics:
1. molecular weight: 576.6
2. vital stain, stain devitalized cells & mucin
3. has antiviral property at 0.06%
4. used as food colorant
Indication: simple screening method for detection of vitamin A deficiency xerophthlamia
S/E: none but longer staining effect
METHYLENE BLUE
LISSAMINE GREEN
 0.1% TRYPAN BLUE
Characteristics:
1. anionic dye
2. stain devitalized cells
S/E: none
Indications:
a. indicator of corneal endothelial integrity for corneal grafting by injecting to A.C.
b. stains anterior lens capsule during phacoemulsification → immature cataract
*The refractive power of the human eye depends on three factors:
1. power of the cornea
2. power of the lens
3. length of the eye
*Following cataract surgery, only the power of the cornea and the length of the eye are
relevant. If both of these variables are known, it is possible to calculate what lens power will give
the best refraction. Biometry is the process of measuring the power of the cornea (keratometry)
and the length of the eye using Optical Biometer. Using this data, it will determine the ideal
intraocular lens (IOL) power to be implanted.
In 10 seconds, six values for cataract surgery are measured:
1. axial length of the eyeball
2. central corneal thickness (CCT)
3. corneal curvature radius
4. anterior chamber depth
5. white to white distance
6. pupil size
After effect of Phacoemulsification: posterior capsule opacity (after cataract)

*The most common complication of adults having standard extracapsular (ECCLE) surgery or
phacoemulsification for cataracts is clouding of the part of the lens covering (capsule) that
remains after surgery, called “posterior capsule opacification or after cataract”. If the cloudiness
affects your vision, you may choose to have a laser surgery called YAG posterior capsulotomy
to correct this problem.
A laser (YAG laser) is used to cut a hole in the clouded back lining of the lens capsule to allow
light to pass through the membrane to the retina at the back of the eye.

Cataract extraction:
1. Extracapsular lens extraction (ECCLE) – removed anterior lens capsule and
cataractous lens
2. Intracapsular cataract Extraction (ICCLE) - removed anterior and posterior
lens capsule and cataractous lens
TRYPAN BLUE
Trypan blue
YAG Laser Capsulotomy
e Corneal Corneal and Corneal Antimicrobial
epithelial defect conjunctival Mucus devitalized property
devitalized cell endothelium

Fluorescein (+) (-) (-) (-)

Fluorexone (+) (+) (+) (-)

Rose Bengal (-) (+) (+) (-) Anti-viral

Indocyanine (-) (-) (-) (+)

Methylene blue (-) (+) (+) (-) Anti-bacterial

Trypan blue (-) (+) (-) (+)

Lissamine (-) (+) (+) (-) Anti-viral

Argyrols (-) (+) (+) (-)