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Imaging microscopic pigment chemistry in conjunctival melanocytic lesions using pump-probe laser microscopy.

Publication ,  Journal Article
Wilson, JW; Vajzovic, L; Robles, FE; Cummings, TJ; Mruthyunjaya, P; Warren, WS
Published in: Invest Ophthalmol Vis Sci
October 21, 2013

PURPOSE: To report the application of a novel imaging technique, pump-probe microscopy, to analyze patterns of pigment chemistry of conjunctival melanocytic lesion biopsies. METHODS: Histopathologic specimens of eight previously excised conjunctival melanocytic lesions were analyzed with pump-probe microscopy. The technique uses a laser scanning microscope with a two-color pulsed laser source to distinguish hemoglobin, eumelanin, and pheomelanin pigment based on differences in transient excited state and ground state photodynamics. The pump-probe signatures of conjunctival melanins were compared with cutaneous melanins. The distributions of hemoglobin, eumelanin, and pheomelanin were analyzed, and pump-probe images were correlated with adjacent hematoxylin and eosin (H&E)-stained sections. RESULTS: The pump-probe signatures of conjunctival melanins are similar, but not identical to cutaneous melanins. In addition, there are qualitative and quantitative differences in the structure and pigment chemistry of conjunctival benign nevi, primary acquired melanosis of the conjunctiva (PAM), and conjunctival melanomas. The pump-probe images correlated well with histopathologic features observed in the adjacent H&E-stained sections, and provided a label-free means of discerning conjunctival anatomic features and pathologic benign or malignant tissue. CONCLUSIONS: Pump-probe laser microscopy shows promise as an adjuvant diagnostic tool in evaluation of ocular melanocytic lesions based on morphologic correlation with the histopathology results and pigment chemistry. This initial study suggests systematic differences in pigmentation patterns among conjunctival benign nevi, primary acquired melanosis, and melanomas. In addition, pump-probe microscopy has the potential for use as a noninvasive "in vivo" optical biopsy technique to aid clinical and surgical management of conjunctival melanocytic lesions.

Duke Scholars

Published In

Invest Ophthalmol Vis Sci

DOI

EISSN

1552-5783

Publication Date

October 21, 2013

Volume

54

Issue

10

Start / End Page

6867 / 6876

Location

United States

Related Subject Headings

  • Ophthalmology & Optometry
  • Microscopy, Fluorescence, Multiphoton
  • Microscopy, Confocal
  • Melanoma
  • Melanins
  • Lasers
  • Humans
  • Hemoglobins
  • Conjunctival Neoplasms
  • 3212 Ophthalmology and optometry
 

Citation

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Wilson, J. W., Vajzovic, L., Robles, F. E., Cummings, T. J., Mruthyunjaya, P., & Warren, W. S. (2013). Imaging microscopic pigment chemistry in conjunctival melanocytic lesions using pump-probe laser microscopy. Invest Ophthalmol Vis Sci, 54(10), 6867–6876. https://doi.org/10.1167/iovs.13-12432
Wilson, Jesse W., Lejla Vajzovic, Francisco E. Robles, Thomas J. Cummings, Prithvi Mruthyunjaya, and Warren S. Warren. “Imaging microscopic pigment chemistry in conjunctival melanocytic lesions using pump-probe laser microscopy.Invest Ophthalmol Vis Sci 54, no. 10 (October 21, 2013): 6867–76. https://doi.org/10.1167/iovs.13-12432.
Wilson JW, Vajzovic L, Robles FE, Cummings TJ, Mruthyunjaya P, Warren WS. Imaging microscopic pigment chemistry in conjunctival melanocytic lesions using pump-probe laser microscopy. Invest Ophthalmol Vis Sci. 2013 Oct 21;54(10):6867–76.
Wilson, Jesse W., et al. “Imaging microscopic pigment chemistry in conjunctival melanocytic lesions using pump-probe laser microscopy.Invest Ophthalmol Vis Sci, vol. 54, no. 10, Oct. 2013, pp. 6867–76. Pubmed, doi:10.1167/iovs.13-12432.
Wilson JW, Vajzovic L, Robles FE, Cummings TJ, Mruthyunjaya P, Warren WS. Imaging microscopic pigment chemistry in conjunctival melanocytic lesions using pump-probe laser microscopy. Invest Ophthalmol Vis Sci. 2013 Oct 21;54(10):6867–6876.

Published In

Invest Ophthalmol Vis Sci

DOI

EISSN

1552-5783

Publication Date

October 21, 2013

Volume

54

Issue

10

Start / End Page

6867 / 6876

Location

United States

Related Subject Headings

  • Ophthalmology & Optometry
  • Microscopy, Fluorescence, Multiphoton
  • Microscopy, Confocal
  • Melanoma
  • Melanins
  • Lasers
  • Humans
  • Hemoglobins
  • Conjunctival Neoplasms
  • 3212 Ophthalmology and optometry