Fluorescein

Dr Rajvin Samuel

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Fluorescein is a synthetic organic

compound available as a dark orange/red powder soluble in water and alcohol.

 Synthesis:

It can be prepared from phthalic anhydride and resorcinol in the presence of zinc chloride via the Friedel-Crafts reaction.
Other name - Resorcinolphthalein

Medical applications :
In forensics and serology to detect latent blood stains In Microbiology To detect Leptospira strains under

microscopy in Syphilis Serology (FTA-ABS Test)

In dentistry - a dye applied to teeth to reveal plaque

 Fluorescein sodium is used extensively as a

diagnostic tool in the field of ophthalmology. It is applied topically in the form of a drop or it can be injected intravenously to produce a fluorescein angiogram. corneal abrasions, corneal ulcers, herpetic corneal infections, and dry eye. Fluorescein angiography is used to diagnose and categorize macular degeneration, diabetic retinopathy, inflammatory intraocular conditions, and intraocular tumors

Topical fluorescein is useful in the diagnosis of

In ophthalmology
Ophthalmic research :
Intraocular dynamic studies [fluorometry] Tear film drainage studies Penetration to anterior segment structures

Fluorescein angiography [in acquired macular & Retinal vascular patholgies]

Staining techniques Test for dry eye Evaluation of tear drainage Applanation tonometry

Fluorescein angiography
Photographic surveillance of passage of fluorescein

through retinal and choroidal circulation

Principle: A molecule is excited

by light of certain wavelength to a higher energy level ,release photon then comes down to original state 2 ml of 25 % [0r] 5 ml of 10 % solution

 80 % protein bound and 20 % free into circulation

PROCEDURE:

White light from camera flash pass through blue excitation filter [488 nm ] Yellow green barrier filter [533 nm] camera film A red free photograph is taken at 1 second interval 5-25 secs after injection onset stimulates back into

 Phases : 1 . Choroidal

2. 3. 4. 5.

Arterial Arteriovenous [capillary] Venous Late [elimination]

Side effects of Fluorescein:

Temporary yellowing , Orange yellow urine , Nausea,vomiting ,Urticaria , Toxic neuritis ,Local tissue necrosis ,Extravasation into skin ,vasovagal reactions

 Hypofluorescence:

1. Vascular filling defects 2.Blockage by haemorrhage RPE hypertrophy ,Hard - exudates, Naevi ,.

Hyperfluorescence:

Leakage Pooling staining Transmission [window defect]

Staining Techniques:
Seidels test for Wound leak
Corneal Staining for ulcers ,other epithelial

defects

Test for dry eye

Tear break up time
To test for tear film stability ,fluorescein dye is added to the eye and the tear film is observed under the slit lamp while the patient avoids blinking until tiny dry spots develop. The longer it takes, the more stable the tear film. A short tear break-up time is a sign of a poor tear film. Generally, >10 seconds is thought to be normal, 5 to 10 seconds, marginal, and <5 seconds low

Tests for lacrimal drainage

Jones dye test :

To distinguish between Anatomical and functional outflow problems

Instil one drop of fluorescein into the conjunctival

sac Put a cotton bud soaked in anaesthetic in the inferior meatus. If fluorescein is detected after five minutes, the system is patent (positive Primary Jones Test).

 No fluorescein - Negative Primary Jones Test

and the functional obstruction could be anywhere from the punctum to the Valve of Hasner.

Next, wash the excess fluorescein from the

conjunctival sac and syringe. Fluorescein detected, entered the sac - positive Secondary Jones Test and functional obstruction of the nasolacrimal duct. Test, because fluorescein not entered the sac and, thus, there is stenosis of the puncta or canalicular system - Anatomical obstruction.

If No dye after syringing, negative Secondary Jones

Applanation tonometry
The intraocular pressure is inferred from the

force required to flatten (applanate) a constant area of the cornea, as per the Imbert-Fick law. 1.Anaesthetic & fluorescein instilled in conjuntival sac 2.With Cobalt blue filter,brightest illumination and prism advanced until touches apex of cornea 3.A pattern of 2 semicircles one above ,other below the horizontal midline

 4. Dial is rotated to align inner margins of

semicircles to just touch each other 5.Reading on dial multiplied by 10 = Intraocular pressure