Digital video-imaging of leukocyte migration in the iris: Intravital microscopy in a physiological model during the onset of endotoxin-induced uveitis

M. D. Becker, R. Nobiling, Stephen Planck, James (Jim) Rosenbaum

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

The process of inflammation is accompanied by an alteration of leukocyte-endothelial dynamics. Reciprocal changes in the endothelium and the white cell permit the leukocyte to relinquish its normal free-flowing state in order to roll, arrest, and emigrate through the endothelium. Although intravital microscopy is an established method to observe this process, the eye has been under-utilized for this purpose. Iris vasculature can be videophotographed without the artifact of trauma. We used rhodamine 6G in vivo staining of leukocytes from BALB/c mice in a model of inflammation induced by intravitreally injected endotoxin. Digital video technology was used to record observations at baseline, 2 h, and 4 h after the endotoxin injection. Off-line analysis of microhemodynamic parameters established that the percentage of venules exhibiting rolling increased significantly from 4% at baseline to 34% at 2 h and 82% at 4 h after endotoxin injection. We found a marked increase in leukocyte arrest within 4 h (601 ± 119 cells per mm2 vs. 2±1 cells per mm2 in control animals). Although shear stress differs minimally between iris arterioles and venules, both rolling and arrest occurred preferentially in venules indicating that shear stress is not the dominant factor for determining cell adhesion. Compared to previous reports on intravital microscopy, our methodology includes refinements or advantages in visualizing cells that have transmigrated as well as the avoidance of surgical trauma. The resolution and quantifiable nature of this technique are such that the methodology can be applied to repetitive observation of leukocyte-endothelial dynamics during an immune response. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)23-37
Number of pages15
JournalJournal of Immunological Methods
Volume240
Issue number1-2
DOIs
StatePublished - Jun 23 2000

Fingerprint

Uveitis
Iris
Endotoxins
Leukocytes
Venules
Endothelium
Inflammation
Injections
Wounds and Injuries
Arterioles
Artifacts
Observation
Intravital Microscopy
Staining and Labeling
Technology

Keywords

  • Angiography
  • Fluorescence microscopy
  • Imaging techniques
  • Iris
  • Iritis
  • Leukocytes
  • Microcirculation
  • Rhodamine
  • Vascular endothelium

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

@article{e7a7454e8c524ad6915f5d1b08117f08,
title = "Digital video-imaging of leukocyte migration in the iris: Intravital microscopy in a physiological model during the onset of endotoxin-induced uveitis",
abstract = "The process of inflammation is accompanied by an alteration of leukocyte-endothelial dynamics. Reciprocal changes in the endothelium and the white cell permit the leukocyte to relinquish its normal free-flowing state in order to roll, arrest, and emigrate through the endothelium. Although intravital microscopy is an established method to observe this process, the eye has been under-utilized for this purpose. Iris vasculature can be videophotographed without the artifact of trauma. We used rhodamine 6G in vivo staining of leukocytes from BALB/c mice in a model of inflammation induced by intravitreally injected endotoxin. Digital video technology was used to record observations at baseline, 2 h, and 4 h after the endotoxin injection. Off-line analysis of microhemodynamic parameters established that the percentage of venules exhibiting rolling increased significantly from 4{\%} at baseline to 34{\%} at 2 h and 82{\%} at 4 h after endotoxin injection. We found a marked increase in leukocyte arrest within 4 h (601 ± 119 cells per mm2 vs. 2±1 cells per mm2 in control animals). Although shear stress differs minimally between iris arterioles and venules, both rolling and arrest occurred preferentially in venules indicating that shear stress is not the dominant factor for determining cell adhesion. Compared to previous reports on intravital microscopy, our methodology includes refinements or advantages in visualizing cells that have transmigrated as well as the avoidance of surgical trauma. The resolution and quantifiable nature of this technique are such that the methodology can be applied to repetitive observation of leukocyte-endothelial dynamics during an immune response. (C) 2000 Elsevier Science B.V.",
keywords = "Angiography, Fluorescence microscopy, Imaging techniques, Iris, Iritis, Leukocytes, Microcirculation, Rhodamine, Vascular endothelium",
author = "Becker, {M. D.} and R. Nobiling and Stephen Planck and Rosenbaum, {James (Jim)}",
year = "2000",
month = "6",
day = "23",
doi = "10.1016/S0022-1759(00)00165-4",
language = "English (US)",
volume = "240",
pages = "23--37",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Digital video-imaging of leukocyte migration in the iris

T2 - Intravital microscopy in a physiological model during the onset of endotoxin-induced uveitis

AU - Becker, M. D.

AU - Nobiling, R.

AU - Planck, Stephen

AU - Rosenbaum, James (Jim)

PY - 2000/6/23

Y1 - 2000/6/23

N2 - The process of inflammation is accompanied by an alteration of leukocyte-endothelial dynamics. Reciprocal changes in the endothelium and the white cell permit the leukocyte to relinquish its normal free-flowing state in order to roll, arrest, and emigrate through the endothelium. Although intravital microscopy is an established method to observe this process, the eye has been under-utilized for this purpose. Iris vasculature can be videophotographed without the artifact of trauma. We used rhodamine 6G in vivo staining of leukocytes from BALB/c mice in a model of inflammation induced by intravitreally injected endotoxin. Digital video technology was used to record observations at baseline, 2 h, and 4 h after the endotoxin injection. Off-line analysis of microhemodynamic parameters established that the percentage of venules exhibiting rolling increased significantly from 4% at baseline to 34% at 2 h and 82% at 4 h after endotoxin injection. We found a marked increase in leukocyte arrest within 4 h (601 ± 119 cells per mm2 vs. 2±1 cells per mm2 in control animals). Although shear stress differs minimally between iris arterioles and venules, both rolling and arrest occurred preferentially in venules indicating that shear stress is not the dominant factor for determining cell adhesion. Compared to previous reports on intravital microscopy, our methodology includes refinements or advantages in visualizing cells that have transmigrated as well as the avoidance of surgical trauma. The resolution and quantifiable nature of this technique are such that the methodology can be applied to repetitive observation of leukocyte-endothelial dynamics during an immune response. (C) 2000 Elsevier Science B.V.

AB - The process of inflammation is accompanied by an alteration of leukocyte-endothelial dynamics. Reciprocal changes in the endothelium and the white cell permit the leukocyte to relinquish its normal free-flowing state in order to roll, arrest, and emigrate through the endothelium. Although intravital microscopy is an established method to observe this process, the eye has been under-utilized for this purpose. Iris vasculature can be videophotographed without the artifact of trauma. We used rhodamine 6G in vivo staining of leukocytes from BALB/c mice in a model of inflammation induced by intravitreally injected endotoxin. Digital video technology was used to record observations at baseline, 2 h, and 4 h after the endotoxin injection. Off-line analysis of microhemodynamic parameters established that the percentage of venules exhibiting rolling increased significantly from 4% at baseline to 34% at 2 h and 82% at 4 h after endotoxin injection. We found a marked increase in leukocyte arrest within 4 h (601 ± 119 cells per mm2 vs. 2±1 cells per mm2 in control animals). Although shear stress differs minimally between iris arterioles and venules, both rolling and arrest occurred preferentially in venules indicating that shear stress is not the dominant factor for determining cell adhesion. Compared to previous reports on intravital microscopy, our methodology includes refinements or advantages in visualizing cells that have transmigrated as well as the avoidance of surgical trauma. The resolution and quantifiable nature of this technique are such that the methodology can be applied to repetitive observation of leukocyte-endothelial dynamics during an immune response. (C) 2000 Elsevier Science B.V.

KW - Angiography

KW - Fluorescence microscopy

KW - Imaging techniques

KW - Iris

KW - Iritis

KW - Leukocytes

KW - Microcirculation

KW - Rhodamine

KW - Vascular endothelium

UR - http://www.scopus.com/inward/record.url?scp=0034705648&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034705648&partnerID=8YFLogxK

U2 - 10.1016/S0022-1759(00)00165-4

DO - 10.1016/S0022-1759(00)00165-4

M3 - Article

C2 - 10854598

AN - SCOPUS:0034705648

VL - 240

SP - 23

EP - 37

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 1-2

ER -