Trypan blue selective staining using whole blood for internal limiting membrane peeling during macular hole corrective surgery

Cutan Ocul Toxicol. 2009;28(3):114-8. doi: 10.1080/15569520903036430.


Objective: To prevent trypan blue (TB) retention and its potential toxicity by selectively staining the internal limiting membrane (ILM) with the aid of whole blood.

Methods: Twenty-four consecutive patients with macular holes who underwent TB-assisted ILM peeling were enrolled. In the surgeries, autologous whole blood was applied to cover the macular hole, and only the area selectively used for TB staining was exposed. Trypan blue solution (0.75 mg/mL, after being diluted with 5% glucose) was used to stain the exposed macular area in the fluid-filled vitreous cavity. The stained ILM was then completely removed.

Results: Twenty-four macular holes were studied. The ILMs were selectively stained by trypan blue and then removed completely. No visible dye remained on the retina after peeling. The macular holes were closed in 22 eyes (92%) after a single surgery. The mean best-corrected visual acuity improved from 20/140 to 20/62 after surgery. In addition, no fundus changes were observed during the follow-up period.

Conclusion: Selective staining of ILM during TB-assisted membrane peeling could be achieved with whole blood as the adjuvant. Complete removal of stained membranes can prevent TB retention and may further reduce toxicity on retinal pigment epithelium and retina.

Publication types

  • Clinical Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Blood*
  • Coloring Agents / toxicity*
  • Epiretinal Membrane / metabolism
  • Epiretinal Membrane / surgery*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Ophthalmologic Surgical Procedures / methods*
  • Prospective Studies
  • Retinal Perforations / metabolism
  • Retinal Perforations / surgery*
  • Staining and Labeling / methods
  • Trypan Blue / toxicity*


  • Coloring Agents
  • Trypan Blue