Fluorescein is a synthetic organic compound available as a dark orange/red powder soluble in water and alcohol.

It is widely used as a fluorescent tracer for many applications.

Uses of fluorescein in ophthalmology

1.

Fluorescein angiography 2. Applanation tonometry 3. Corneal staining

Fluorescein angiography

Fluorescein angiography is a technique which is based on the detection of fluorescent light emitted by a dye in circulation. Mechanism y It involves injection of sodium fluorescein into the systemic circulation, and then an angiogram is obtained by photographing the fluorescence emitted after illumination of the retina with blue light at a wavelength of 490 nanometers.

Equipment 1. Exciter filter : Allows only blue light to illuminate the retina, depending on the filter the excitation wavelength hitting the retina will be between 465490 nm
2.

Barrier filter: Allows only yellow-green light (from the fluorescence) to reach the camera, depending on the filter it can be between 520-530 nm. Fundus Camera

3.

Technique
y 2-5cc fluorescein dye is injected intravenously y White light from a flash is passed through a blue excitation y

y y y

filter. Blue light is then absorbed by unbound fluorescein molecules, emitting light with a longer wavelength in the yellow-green spectrum (520-530nm). Barrier filter blocks any reflected light so that the images capture only light emitted from the fluorescein Images are acquired immediately after injection and continue for ten minutes depending on the pathology being imaged. These images are recorded digitally or on 35mm film.

Normal circulatory filling

y 0 seconds y y y y y y y y

injection of fluorescein 9.5 sec posterior cilliary arteries 10 sec choroidal flush 10-12 sec retinal arterial stage 13 sec capillary transition stage 14-15 sec early venous stage or lamellar stage 16-17 sec venous stage 18-20 sec late venous stage 5 minutes late staining

Uses:1.

Exposing depth pathological involvement in diabetic retinopathy and reveal neovascularisatio0n occurring in any area of fundus Assessment of disorder of fundus including neoplasia, disorder of optic nerve head such as papilloedema Interpretation of neovascularisation of the iris when leakage from vessels of the iris Vitreous fluorophotometry - allow measurement of fludescein concentration in all parts of vitreous chamber visible through eyepiece of the slitlamp

2.

3.

4.

Pathological findings
Causes of hyper fluorescence : 1. leaking defects(choroidal or diabetic neovascularization ) 2. pooling defects 3. staining 4. abnormal vasculature

Causes of hypo fluorescence : 1. blocking defect (corneal scar, cataract, vitreous hemorrhage, 2. filling defect (retinal or choroidal vascular occlusion )

Complication:1. 2. 3. 4. 5. 6.

Nausea Vomiting Pruritis Pyrexia Thrombophlebitis Local tissue necrosis can occur with extravasation of dye

Applanation Tonometry
Applanation tonometry is technique which is use to measures intraocular pressure either by the force required to flatten a constant area of the cornea (e.g. Goldmann tonometry) or by the area flattened by a constant force.

Equipment
y Tonometer, either Goldmann (used on slit

lamps) or Perkins (hand-held) y Applanation prism y Local anaesthetic drops y Fluorescein strips y Clean cotton wool or gauze swabs.

Method
y Instil the local anaesthetic drops and then the fluorescein y For measuring the IOP in the right eye , the slit beam which is

y y y y y y

shining onto the tonometer head should be from the patient s right side Move the filters so that the blue filter is used to produce a blue beam The beam of light should be as wide as possible and bright as possible. This makes visualising the fluorescein rings easier (with the slit diaphragm fully open) Ask the patient to look straight ahead, open both eyes wide, fix his or her gaze and keep perfectly still With the thumb, gently hold up the patient s top eyelid, taking care not to put any pressure on the eye Direct the blue light from the slit lamp onto the prism head ,Make sure that the tonometer head is perpendicular to the eye

y Move the tonometer forward slowly until the prism rests

gently on the centre of the patient s cornea y With the other hand, turn the calibrated dial on the tonometer clockwise until the two fluorescein semi-circles in the prism head are seen to meet and form a horizontal S shape

Finding:

Purpose
y To help diagnose glaucoma or high eye pressure. y The test is often part of a routine eye exam.

Factors affecting results y An irregularly shaped cornea Interpretation y Pressure readings are in millimetres (mm) of mercury (Hg). A normal reading is about 20 mm Hg or lower. Higher readings may indicate either glaucoma or ocular hypertension.

Advantages
y It's a quick, easy test for glaucoma. y It's non-invasive.

Disadvantages
y Other tests may be necessary to diagnose glaucoma; these

include visual field tests and ophathalmoscopy to evaluate the optic nerve.

Corneal staining
This is a test that uses fluorescein and a blue light to detect foreign bodies in the eye. This test can also detect damage to the cornea, the outer surface of the eye.

Technique:A piece of blotting paper containing the dye will be touched to the surface of patient eye and asked to blink. A blue light is then directed to eye. Any problems on the surface of the cornea will be stained by the dye and appear green under the blue light. Depending on the size, location, and shape of the staining it can determine cause cornea problem

Normal Results
y If the test result is normal, the dye remains in the tear film

on the surface of the eye and does not adhere to the eye itself.

Abnormal Results
y Abnormal tear production (dry eye) y Corneal abrasion(a scratch on the surface of the cornea) y Foreign bodies, such as eyelashes or dust (see eye - foreign

object in) y Infection y Injury or trauma y Severe dry eye associated with arthritis (keratoconjunctivitis sicca)

Disease cornea with stain