Methyl blue and aniline blue versus patent blue and trypan blue as vital dyes in cataract surgery: capsule staining properties and cytotoxicity to human cultured corneal endothelial cells

J Cataract Refract Surg. 2011 Jun;37(6):1147-53. doi: 10.1016/j.jcrs.2010.12.051.


Purpose: To evaluate capsule-staining properties and biocompatibility of the triarylmethane dyes methyl blue and aniline blue compared with patent blue and trypan blue on cultured human corneal endothelial cells.

Setting: Centre for Ophthalmology, University of Tuebingen, Tuebingen, Germany.

Design: Experimental study.

Methods: Human corneal endothelial cell cultures were harvested from human donor cells and exposed to various concentrations (0.025 to 5.0 mg/mL) of methyl blue, aniline blue, patent blue, and trypan blue. Cytotoxicity was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide test after 24 hours of incubation. Calcein live cell staining was performed at the same time point. The dyes were also used to stain pig lens capsules in vitro by incubating the lenses for 1 minute with 3 concentrations (0.5, 1.5, and 2.5 mg/mL) of dye, after which the staining properties were evaluated.

Results: No significant cytotoxicity was detected for patent blue and methyl blue at any tested concentration. However, aniline blue exerted significant cytotoxicity at concentrations of 1.5 mg/mL or higher and trypan blue at 2.5 mg/mL or higher. Capsule staining of the tested triarylmethane dyes was suitable for performing capsulorhexis, but only at higher concentrations than with trypan blue.

Conclusions: High concentrations and long incubation times of trypan blue and aniline blue showed significant cytotoxicity to human cultured endothelial cells in contrast to patent blue and methyl blue. All tested dyes were able to stain lens capsules sufficiently for capsulorhexis creation.

Financial disclosure: No author has a financial or proprietary interest in any material or method mentioned.

Publication types

  • Comparative Study

MeSH terms

  • Aniline Compounds / toxicity
  • Animals
  • Benzenesulfonates / toxicity
  • Cataract Extraction*
  • Cells, Cultured
  • Coloring Agents / toxicity*
  • Endothelium, Corneal / drug effects*
  • Endothelium, Corneal / metabolism
  • Fluoresceins / metabolism
  • Formazans
  • Humans
  • Lens Capsule, Crystalline / anatomy & histology*
  • Rosaniline Dyes / toxicity
  • Staining and Labeling / methods
  • Swine
  • Tetrazolium Salts
  • Trypan Blue / toxicity


  • Aniline Compounds
  • Benzenesulfonates
  • Coloring Agents
  • Fluoresceins
  • Formazans
  • Rosaniline Dyes
  • Tetrazolium Salts
  • MTT formazan
  • aniline blue
  • Methyl Blue
  • sulfan blue
  • Trypan Blue
  • fluorexon