Combined Staining Techniques for Demonstration of Staphylococcus aureus Biofilm in Routine Histopathology
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Combined Staining Techniques for Demonstration of Staphylococcus aureus Biofilm in Routine Histopathology. / Jensen, Louise Kruse; Henriksen, Nicole Lind; Bjarnsholt, Thomas; Kragh, Kasper Nørskov; Jensen, Henrik Elvang.
In: Journal of Bone and Joint Infection, Vol. 3, No. 1, 2018, p. 27-36.Research output: Contribution to journal › Journal article › peer-review
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TY - JOUR
T1 - Combined Staining Techniques for Demonstration of Staphylococcus aureus Biofilm in Routine Histopathology
AU - Jensen, Louise Kruse
AU - Henriksen, Nicole Lind
AU - Bjarnsholt, Thomas
AU - Kragh, Kasper Nørskov
AU - Jensen, Henrik Elvang
PY - 2018
Y1 - 2018
N2 - Aim: Visualization of Staphylococcus aureus biofilm using histochemical staining and combined histochemistry (HC) and immunohistochemistry (IHC).Methods: The ability of S. aureus S54F9 to form biofilm was tested in vitro. Hereafter, infected bone tissue was collected from two different porcine models of osteomyelitis inoculated with S. aureus strain S54F9. The infection time was five and fifteen days, respectively. Twenty-five different histochemical staining protocols were tested in order to find the stains that could identify extracellular biofilm matrix. Protocols with an optimal visualization of biofilm extracellular matrix were combined with an immunohistochemical protocol based on a specific antibody against S. aureus. The combined protocols were applied to the tissue from the porcine models and to infected bone tissue from a child suffering from chronic staphylococcal osteomyelitis for more than a year.Results: S. aureus S54F9 showed an ability to form biofilm in vitro. Visualization of biofilm, i.e. bacterial cells and extracellular matrix in different colours, was seen when the immunohistochemical protocol was combined with Alcian Blue pH3, Luna and Methyl-pyronin green. The bacterial cells were red to light brown and the extracellular matrix either light blue, blue or orange depending on the histochemical stain. In the porcine models and the human case 10 and 90 percent, respectively, of the bacterial aggregates in a 100x magnification field displayed both the extracellular matrix and the bacterial cells simultaneously in two different colours.Conclusions: A combination of HC and IHC can be used to diagnose and characterise biofilm infections on a routine basis.
AB - Aim: Visualization of Staphylococcus aureus biofilm using histochemical staining and combined histochemistry (HC) and immunohistochemistry (IHC).Methods: The ability of S. aureus S54F9 to form biofilm was tested in vitro. Hereafter, infected bone tissue was collected from two different porcine models of osteomyelitis inoculated with S. aureus strain S54F9. The infection time was five and fifteen days, respectively. Twenty-five different histochemical staining protocols were tested in order to find the stains that could identify extracellular biofilm matrix. Protocols with an optimal visualization of biofilm extracellular matrix were combined with an immunohistochemical protocol based on a specific antibody against S. aureus. The combined protocols were applied to the tissue from the porcine models and to infected bone tissue from a child suffering from chronic staphylococcal osteomyelitis for more than a year.Results: S. aureus S54F9 showed an ability to form biofilm in vitro. Visualization of biofilm, i.e. bacterial cells and extracellular matrix in different colours, was seen when the immunohistochemical protocol was combined with Alcian Blue pH3, Luna and Methyl-pyronin green. The bacterial cells were red to light brown and the extracellular matrix either light blue, blue or orange depending on the histochemical stain. In the porcine models and the human case 10 and 90 percent, respectively, of the bacterial aggregates in a 100x magnification field displayed both the extracellular matrix and the bacterial cells simultaneously in two different colours.Conclusions: A combination of HC and IHC can be used to diagnose and characterise biofilm infections on a routine basis.
U2 - 10.7150/jbji.22799
DO - 10.7150/jbji.22799
M3 - Journal article
C2 - 29545993
VL - 3
SP - 27
EP - 36
JO - Journal of Bone and Joint Infection
JF - Journal of Bone and Joint Infection
SN - 2206-3552
IS - 1
ER -
ID: 192741486