Computer-assisted digital image analysis to quantify the mass and purity of isolated human islets before transplantation.
Details
Serval ID
serval:BIB_7CAA94248BDC
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Computer-assisted digital image analysis to quantify the mass and purity of isolated human islets before transplantation.
Journal
Transplantation
ISSN
1534-6080 (Electronic)
ISSN-L
0041-1337
Publication state
Published
Issued date
15/12/2008
Peer-reviewed
Oui
Volume
86
Number
11
Pages
1603-1609
Language
english
Notes
Publication types: Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Publication Status: ppublish
Abstract
Accurate determination of islet purity and mass before transplantation is an essential part of quality control. The standard method is based on manual evaluation of these parameters and thus subjective and prone to errors. Therefore, we developed a computerized approach aimed at evaluating more objectively the number and purity of isolated human islets.
Islets were isolated and purified from human pancreata according to a standard method. For each preparation, two samples were dithizone stained. One sample was analyzed manually by microscopy, following the standard procedure, and the other was digitally photographed for both digital manual and computerized analyses. Computerized analysis was performed using the MetaMorph and ImageJ softwares to automatically quantify purity and size of islets. Islet equivalent (IEQ) number was calculated using the Ricordi algorithm or considering the individual volume of each islet. Computerized analysis was validated using calibrated red glass microspheres.
When digital manual and computerized analyses were compared, mean values of total islet number, IEQ number calculated using the Ricordi algorithm, and purity were similar. Comparisons of individual values showed good correlations (r>or=0.89). By standard manual analysis, total islet number and purity were higher and IEQ number similar compared with digital manual and computerized analyses. IEQ number was 10% lower (P<0.0001) when calculated using individual sphere volumes compared with the Ricordi algorithm. Measurement of red glass microspheres showed identical values comparing standard manual and computerized analyses.
Computer-assisted digital image analysis is an objective and a reliable method for analyzing pancreatic islets before transplantation.
Islets were isolated and purified from human pancreata according to a standard method. For each preparation, two samples were dithizone stained. One sample was analyzed manually by microscopy, following the standard procedure, and the other was digitally photographed for both digital manual and computerized analyses. Computerized analysis was performed using the MetaMorph and ImageJ softwares to automatically quantify purity and size of islets. Islet equivalent (IEQ) number was calculated using the Ricordi algorithm or considering the individual volume of each islet. Computerized analysis was validated using calibrated red glass microspheres.
When digital manual and computerized analyses were compared, mean values of total islet number, IEQ number calculated using the Ricordi algorithm, and purity were similar. Comparisons of individual values showed good correlations (r>or=0.89). By standard manual analysis, total islet number and purity were higher and IEQ number similar compared with digital manual and computerized analyses. IEQ number was 10% lower (P<0.0001) when calculated using individual sphere volumes compared with the Ricordi algorithm. Measurement of red glass microspheres showed identical values comparing standard manual and computerized analyses.
Computer-assisted digital image analysis is an objective and a reliable method for analyzing pancreatic islets before transplantation.
Keywords
Algorithms, Cell Separation/methods, Glass, Humans, Image Processing, Computer-Assisted/methods, Islets of Langerhans/cytology, Islets of Langerhans/metabolism, Islets of Langerhans Transplantation/instrumentation, Islets of Langerhans Transplantation/methods, Microscopy, Microspheres, Pancreas/pathology, Quality Control, Reproducibility of Results, Software, Tissue and Organ Procurement/methods
Pubmed
Web of science
Open Access
Yes
Create date
14/06/2021 9:59
Last modification date
24/05/2024 12:40