Biophysical and morphological evaluation of human normal and dry eye meibum using hot stage polarized light microscopy

Igor A. Butovich, Hua Lu, Anne McMahon, Howard Ketelson, Michelle Senchyna, David Meadows, Elaine Campbell, Mike Molai, Emily Linsenbardt

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Purpose. To study melting characteristics and the morphology of human and mouse meibum. Methods. Hot stage cross-polarized light microscopy (HSPM) and immunohistochemical approaches were used. Results. Isolated human meibum, and meibum of mice (either isolated or within the meibomian ducts of mice), were found to be in liquid-crystal state at physiological temperatures. Melting of both types of meibum started at approximately 10°C and was completed at approximately 40°C. Melting curves of isolated meibum and meibum inside the meibomian ducts were multiphasic with at least two or three clearly defined phase transition temperatures, typically at approximately 12 ± 2°C (minor transition), 21 ± 3°C, and 32 ± 3°C, regardless the source of meibum. Melting was highly cooperative in nature. Samples of abnormal human meibum collected from dry eye patients with meibomian gland dysfunction often showed an increased presence of nonlipid, nonmelting, nonbirefringent, chloroform-insoluble inclusions of a protein nature. The inclusions were positively stained for cytokeratins. The presence of these inclusions was semiquantitatively characterized using a newly proposed 0 to 4 scale. In the presence of large amounts of these inclusions, melting characteristics of meibum and its structural integrity were altered. Conclusions. HSPM is an effective tool that is suitable for biophysical and morphological evaluation of meibum. Morphological properties and melting characteristics of human meibum were found to be similar to those of mice. Abnormal meibum of many dry eye patients contained large quantities of nonlipid, protein-like inclusions, which were routinely absent in meibum of normal controls.

Original languageEnglish (US)
Pages (from-to)87-101
Number of pages15
JournalInvestigative Ophthalmology and Visual Science
Volume55
Issue number1
DOIs
StatePublished - Nov 26 2013

Fingerprint

Polarization Microscopy
Freezing
Meibomian Glands
Liquid Crystals
Transition Temperature
Phase Transition
Chloroform
Keratins
Proteins
Temperature

Keywords

  • Biophysics
  • Hot stage polarized light microscopy
  • Lipids
  • Meibomian glands
  • Phase transitions
  • Rheology

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Biophysical and morphological evaluation of human normal and dry eye meibum using hot stage polarized light microscopy. / Butovich, Igor A.; Lu, Hua; McMahon, Anne; Ketelson, Howard; Senchyna, Michelle; Meadows, David; Campbell, Elaine; Molai, Mike; Linsenbardt, Emily.

In: Investigative Ophthalmology and Visual Science, Vol. 55, No. 1, 26.11.2013, p. 87-101.

Research output: Contribution to journalArticle

Butovich, Igor A. ; Lu, Hua ; McMahon, Anne ; Ketelson, Howard ; Senchyna, Michelle ; Meadows, David ; Campbell, Elaine ; Molai, Mike ; Linsenbardt, Emily. / Biophysical and morphological evaluation of human normal and dry eye meibum using hot stage polarized light microscopy. In: Investigative Ophthalmology and Visual Science. 2013 ; Vol. 55, No. 1. pp. 87-101.
@article{c1746b97fffa4c5fa12762b12d12d637,
title = "Biophysical and morphological evaluation of human normal and dry eye meibum using hot stage polarized light microscopy",
abstract = "Purpose. To study melting characteristics and the morphology of human and mouse meibum. Methods. Hot stage cross-polarized light microscopy (HSPM) and immunohistochemical approaches were used. Results. Isolated human meibum, and meibum of mice (either isolated or within the meibomian ducts of mice), were found to be in liquid-crystal state at physiological temperatures. Melting of both types of meibum started at approximately 10°C and was completed at approximately 40°C. Melting curves of isolated meibum and meibum inside the meibomian ducts were multiphasic with at least two or three clearly defined phase transition temperatures, typically at approximately 12 ± 2°C (minor transition), 21 ± 3°C, and 32 ± 3°C, regardless the source of meibum. Melting was highly cooperative in nature. Samples of abnormal human meibum collected from dry eye patients with meibomian gland dysfunction often showed an increased presence of nonlipid, nonmelting, nonbirefringent, chloroform-insoluble inclusions of a protein nature. The inclusions were positively stained for cytokeratins. The presence of these inclusions was semiquantitatively characterized using a newly proposed 0 to 4 scale. In the presence of large amounts of these inclusions, melting characteristics of meibum and its structural integrity were altered. Conclusions. HSPM is an effective tool that is suitable for biophysical and morphological evaluation of meibum. Morphological properties and melting characteristics of human meibum were found to be similar to those of mice. Abnormal meibum of many dry eye patients contained large quantities of nonlipid, protein-like inclusions, which were routinely absent in meibum of normal controls.",
keywords = "Biophysics, Hot stage polarized light microscopy, Lipids, Meibomian glands, Phase transitions, Rheology",
author = "Butovich, {Igor A.} and Hua Lu and Anne McMahon and Howard Ketelson and Michelle Senchyna and David Meadows and Elaine Campbell and Mike Molai and Emily Linsenbardt",
year = "2013",
month = "11",
day = "26",
doi = "10.1167/iovs.13-13355",
language = "English (US)",
volume = "55",
pages = "87--101",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "1",

}

TY - JOUR

T1 - Biophysical and morphological evaluation of human normal and dry eye meibum using hot stage polarized light microscopy

AU - Butovich, Igor A.

AU - Lu, Hua

AU - McMahon, Anne

AU - Ketelson, Howard

AU - Senchyna, Michelle

AU - Meadows, David

AU - Campbell, Elaine

AU - Molai, Mike

AU - Linsenbardt, Emily

PY - 2013/11/26

Y1 - 2013/11/26

N2 - Purpose. To study melting characteristics and the morphology of human and mouse meibum. Methods. Hot stage cross-polarized light microscopy (HSPM) and immunohistochemical approaches were used. Results. Isolated human meibum, and meibum of mice (either isolated or within the meibomian ducts of mice), were found to be in liquid-crystal state at physiological temperatures. Melting of both types of meibum started at approximately 10°C and was completed at approximately 40°C. Melting curves of isolated meibum and meibum inside the meibomian ducts were multiphasic with at least two or three clearly defined phase transition temperatures, typically at approximately 12 ± 2°C (minor transition), 21 ± 3°C, and 32 ± 3°C, regardless the source of meibum. Melting was highly cooperative in nature. Samples of abnormal human meibum collected from dry eye patients with meibomian gland dysfunction often showed an increased presence of nonlipid, nonmelting, nonbirefringent, chloroform-insoluble inclusions of a protein nature. The inclusions were positively stained for cytokeratins. The presence of these inclusions was semiquantitatively characterized using a newly proposed 0 to 4 scale. In the presence of large amounts of these inclusions, melting characteristics of meibum and its structural integrity were altered. Conclusions. HSPM is an effective tool that is suitable for biophysical and morphological evaluation of meibum. Morphological properties and melting characteristics of human meibum were found to be similar to those of mice. Abnormal meibum of many dry eye patients contained large quantities of nonlipid, protein-like inclusions, which were routinely absent in meibum of normal controls.

AB - Purpose. To study melting characteristics and the morphology of human and mouse meibum. Methods. Hot stage cross-polarized light microscopy (HSPM) and immunohistochemical approaches were used. Results. Isolated human meibum, and meibum of mice (either isolated or within the meibomian ducts of mice), were found to be in liquid-crystal state at physiological temperatures. Melting of both types of meibum started at approximately 10°C and was completed at approximately 40°C. Melting curves of isolated meibum and meibum inside the meibomian ducts were multiphasic with at least two or three clearly defined phase transition temperatures, typically at approximately 12 ± 2°C (minor transition), 21 ± 3°C, and 32 ± 3°C, regardless the source of meibum. Melting was highly cooperative in nature. Samples of abnormal human meibum collected from dry eye patients with meibomian gland dysfunction often showed an increased presence of nonlipid, nonmelting, nonbirefringent, chloroform-insoluble inclusions of a protein nature. The inclusions were positively stained for cytokeratins. The presence of these inclusions was semiquantitatively characterized using a newly proposed 0 to 4 scale. In the presence of large amounts of these inclusions, melting characteristics of meibum and its structural integrity were altered. Conclusions. HSPM is an effective tool that is suitable for biophysical and morphological evaluation of meibum. Morphological properties and melting characteristics of human meibum were found to be similar to those of mice. Abnormal meibum of many dry eye patients contained large quantities of nonlipid, protein-like inclusions, which were routinely absent in meibum of normal controls.

KW - Biophysics

KW - Hot stage polarized light microscopy

KW - Lipids

KW - Meibomian glands

KW - Phase transitions

KW - Rheology

UR - http://www.scopus.com/inward/record.url?scp=84891882278&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84891882278&partnerID=8YFLogxK

U2 - 10.1167/iovs.13-13355

DO - 10.1167/iovs.13-13355

M3 - Article

C2 - 24282231

AN - SCOPUS:84891882278

VL - 55

SP - 87

EP - 101

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 1

ER -