April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Improving 3D mucin visualization by a new device to make impression cytology comparing normal and muco-deficient subjects
Author Affiliations & Notes
  • Basilio Colligris
    Bioquimica y Biologia Molecular IV, Universidad Complutense De Madrid, Madrid, Spain
  • Alba Martin-Gil
    Bioquimica y Biologia Molecular IV, Universidad Complutense De Madrid, Madrid, Spain
  • Begoña Fonseca
    Bioquimica y Biologia Molecular IV, Universidad Complutense De Madrid, Madrid, Spain
  • Gonzalo Carracedo
    Optica II, Universidad Complutense De Madrid, Madrid, Spain
  • Jesus Pintor
    Bioquimica y Biologia Molecular IV, Universidad Complutense De Madrid, Madrid, Spain
  • Footnotes
    Commercial Relationships Basilio Colligris, None; Alba Martin-Gil, None; Begoña Fonseca, None; Gonzalo Carracedo, None; Jesus Pintor, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 4874. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Basilio Colligris, Alba Martin-Gil, Begoña Fonseca, Gonzalo Carracedo, Jesus Pintor, ; Improving 3D mucin visualization by a new device to make impression cytology comparing normal and muco-deficient subjects. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4874.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: The aim of this study was to validate the use of a new medical device to make impression cytology with a new mucin visualization technique allowing not only assess Goblet cells density, but to quantify mucins release in the ocular conjunctiva.

Methods: In this study were taken conjunctival impression cytology samples from 25 healthy subjects (25 eyes) and samples from 29 subjects (29 eyes) with some kind of muco-deficient pathology. It was used the Eyeprim TM device that requires no topical anesthesia and allows collection of 50 % more cells than the traditional method. Treatment of cytology samples was done by PAS- hematoxylin. For measuring cell density, the display of cytology by impression was made by transmitted light microscopy. For the new method of displaying mucins in 3D, the samples were analyzed with a confocal laser microscope (LSM) by which the different sample dyes were excited at 488 and 543 nm. For each image were made a series of optical slides in axis z, that were reconstructed to obtain a three dimensional image.

Results: With this new technique the Goblet cells are easily distinguishable from the epithelial cells, but also the 3D image permits to measure new parameters, such as the height of the mucin cloud secreted by each cell ( 8.81 ± 4.00 microns ), the mucin diffusion of the epithelial cells ( 2.77 ± 1.00 microns ) and the thickness of the Goblet cell. These parameters were significantly decreased, by about 70 % and 40% respectively, in the group of muco-deficient patients.

Conclusions: The mucin visualization technique by confocal laser microscopy in 3D is a solid diagnostic tool, providing a number of parameters related to both Goblet cells secretion and the cell itself, allowing more objective assessments of the mucin secretion status, than the classical measure of cell density. The use of the new medical device for impression cytology EyePrim improves the features of 3D LSM analysis.

Keywords: 596 microscopy: confocal/tunneling • 491 cytology • 485 cornea: surface mucins  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×