Fluorescein stain is a commonly used diagnostic agent in ophthalmology. It is a water-soluble yellow dye that appears bright green when illuminated by blue light if there is a defect or break in the corneal epithelium or conjunctiva. It has various applications including assessing ocular surface integrity, fitting contact lenses, measuring intraocular pressure, and evaluating dry eye and lacrimal drainage. While generally safe, it can cause allergic reactions through topical or intravenous use. Proper sterilization of fluorescein drops is important to prevent bacterial contamination.

1. USED IN OPHTHALMOLOGY
Indra P Sharma
Master of Clinical Optometry (Year I)
Amity Medical School
FLUORESCEIN STAIN

2. Objective
 To understand the biochemical
properties, indication and
contraindication of fluorescein stain
ophthalmology.
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3. Contents
1. An overview- Introduction
2. Properties of sodium fluorescein
3. Indication of use
4. Contraindication, side effects and
comtamination
5. Conculsion
6. Reference
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4. An overview
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5. Introduction –Sodium
Fluorescein
 Fluorescein is a
synthetic organic
compound available as a
dark orange/ yellow water-
soluble dibasic dye of
xanthine series.
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 Soduim Fluorescein - one of the most useful
and most commonly used diagnostic agents
(stains) in ophthalmology and optometry.

6. Fluorescein – Historical
Perspective
 Baeyer(1871): First fluorescein dye was
made.
 M Straub (1888) : First used fluorescein for
vital staining of the eye.
 Burk (1910): First used fluorescein to detect
retinal disease
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7. Properties of fluorescein
 A yellow water-soluble dibasic dye
of xanthine series
 Orange red in powder and yellow
in solution.
 Chemical formula: C2H12O5Na
 Molecular weight: 376.27
 Solubility : 50% (in water at 15 C)

8. Optimum condition for
observation of fluorescein
 For dilute concentrations of fluorescein in an
aqueous solution
 Peak absorption:wavelength between 485 and 500 nm
 Peak emission: wavelength between 525 and 530nm
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 The fluorescent light appears
yellow green in blue light.
 The flourescence increases with
greater concentration upto
0.001% and greater pH upto 8.

9. Important clinical
characterstics
 Stains epithelial defects bright green
 Diffuses into intercellular space
 Will not stain devitalized
 Tear film appears yellow orange
 Can exhibit pseudoflare, Fischer Schweitzer mosaic
 Promotes growth of pseudomonas aeruginosa in
solution
 Will stain soft contact lens
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10. Preparation for tropical
ocular use
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11. Available forms
 Can be applied to eye
 Topically in form of
solution
 By Fluorescein
impregnated filter paper
strips (developed by
kimura)
 Injectable form for IV use

12. Indication for use
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13. A. Topical Indication
 Assessment of ocular surface integrity -
Detection of defects in corneal epithelium
 Fitting assessment of rigid contact lens.
 Applanation tonometry - Goldmann
tonometer/Perkins hand-held tonometry
 Seidel's test- Detection of site of perforation/bleb
 Lacrimal testing ( Tear flim breakup time
(TBUT), Jone dye test, Fluorescein dye
disappreance test(FDDT)
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14. 1.Assessment of ocular surface
integrity
 Frequently used to detect lesions of ocular
surface owing to its high degree of ionization,
it neither penetrates the intact corneal
epithelium nor forms a firm bond with any vital
tissue.
 Instillation of dye in cul-de-sac allows
determination of corneal & conjunctival lesions
such as abrasions ulcers& edema & aids in
detection of foreign bodies.
 Epithelial defect appears as vivid green
fluorescence
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15. How does staining take
place?
Any break in
epithelium
Penetration of
Fluorescein in
adjoining
bowman’s &
stromal layer
Dye makes
contact
with an
alkaline
interstitial
fluid
Fluid turns bright
green owing to its
PH indicator
properties &
depending to
extent of lesion
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16. Staining of corneal infiltrate
Corneal abrasionSharma IP Conjunctival lesion

17. 2. Seidel's test
 Detection of site of perforation/bleb
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18.  A major aid in fitting of RGP contact lenses is
vital staining of tear film
 Observation of Fluorescein stained tear film
with a cobalt filter of slit lamp allows
determination of the fit of lens
 Useful in assessing the integrity of cornea in
CL users as the dye can disclose areas where
the CL disrupts the corneal epithelium
3.Contact lens fitting and
management
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19. Stained eye with contact lens
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20. 5.Evaluation for dry eye & lacrimal
system
 Topically applied Fluorescein –used to
evaluate integrity of the precorneal tear film&
patency of the lacrimal drainage system
 Assessment of TBUT
 Evaluating the EPIPHORA
 Assessment of FDDT
 To distinguish between
Anatomical and functional
outflow problems-JONES DYE TEST
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22. 4.Applanation tonometry
 Important component in measuring IOP with
Goldmann applanation tonometer
 Requires the meniscus of tear fluid surrounding
the flattened corneal surface be sufficiently
stained so that apex of the wedge shaped
meniscus is visible.
 Procedure
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 midlineSharma IP

23. Sharma IP

24. B. Intravenous indication
1. Fluorescein angiography
 About 10 ml of a 5% solution injected in
antecubital vein
 The dye normally appears in central retinal artery
in 10– 15 sec
 Shows retinal blood vessels in high contrast
 Non vascularised, pigmented retinal & subretinal
lesions appear as dark areas against the green
fluorescing background
 Proven helpful in diagnosis of a variety of
pathological conditions of fundus ,various macular
lesions , choroidopathy, diabetic retinopathy etc.
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25. Sharma IP

26. Clinical photographs after fluorescein
angiographySharma IP

27. 2.Iris Angiography
 IV inj. Dye first appears in radial vessels
at betn 9-20 secs
 Amount of iris pigmentation and the pattern of
its distribution compared with normal iris
angiogram
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28. Ophthalmic Research
 Intraocular dynamic studies [fluorometry]
 Tear film drainage studies
 Penetration to anterior segment structures
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29. Side effects
 Topical - unconsciousness & hypertensive
reactions
 Rare side effects.
 IV inj. – with increased concentration adverse
effects in about 10% of patients receiving IV
inj.
 Less frequently – respiratory effects like
laryngeal or pulmonary edema
 Cardiovascular toxicity in form of severe
hypotension and shock
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30. Contraindication
 Hypersensitivity to active ingredents or any
other components
 Family and personal allergic history
 Not used over soft contact lens.
SCL – avoided for few hrs. of fluorescein
instillation
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31. Contamination of fluorescein
 Contamination of fluorescein eyedrops is a serious
risk
-even greater than that encountered with the majority
of other eyedrops.
 As these individual drops are liable to become
infected with bacteria and, at the same time, are
frequently used on damaged tissue that is prone to
infection, very great care must be taken in their use.
 Pseudomonas aeruginosa – most dangerous
microorganism with which fluorescein eyedrops are
inclined to become invaded.
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32. Contd...
 Phenylmercuric acetate or nitrate in
0.002%
-Best bactericide for preserving
fluorescein drops, and this is effective
against Pseudomonas, given adequate
contact time.
 However, the safest method is sterile
single-dose units or sterile fluorescein-
impregnated paper strips, both are
readily available and to be highly
recommended.Sharma IP

33. Conclusion
 Fluorescein stain is a very useful diagnostic
agents in ophthalmic clinical practise and has
many applications.
 The use of diagnostic dyes represents one of the
most efficient, objective, non-invasive, and directly
visible means we have of identifying and tracking
ocular structures at the cellular level.
 Every optometrist must understand the proper use
of its clinical application, contraindication and side
effects in clinical use.
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34. Reference
Books
 Donald S. Fong,Drugs in Ophthalmology,,2006, Springer-
Verlag Berlin Heidelberg
 Graham Hopkins and Richard Pearson, Ophthalmic Drugs,
2007, Butterworth Heineman Elsevier. 5th Ed10:149-154
 Brain Duvall, Ophthalmic medication and Pharmacology,
SLACK incorporated 2nd Ed.
 P.H.O’Connor Davies, The Action and Uses of Ophthalmic
Drugs,1994, Jaypee Brothers. 3rd Ed. 9:148-153
Websites
 www.emedicine.medscape.com
 www.rootatlas.com
 en.wikipedia.org
 www.google.com/imghp
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35. Tashi Delek
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