Limits...
Endogenous Two-Photon Excited Fluorescence Provides Label-Free Visualization of the Inflammatory Response in the Rodent Spinal Cord.

Uckermann O, Galli R, Beiermeister R, Sitoci-Ficici KH, Later R, Leipnitz E, Neuwirth A, Chavakis T, Koch E, Schackert G, Steiner G, Kirsch M - Biomed Res Int (2015)

Bottom Line: Iba1-positive microglia were found in areas lacking any TPEF signal.Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF.Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes.

View Article: PubMed Central - PubMed

Affiliation: Neurosurgery, Carl Gustav Carus University Hospital, TU Dresden, Fetscherstraße 74, 01307 Dresden, Germany.

ABSTRACT
Activation of CNS resident microglia and invasion of external macrophages plays a central role in spinal cord injuries and diseases. Multiphoton microscopy based on intrinsic tissue properties offers the possibility of label-free imaging and has the potential to be applied in vivo. In this work, we analyzed cellular structures displaying endogenous two-photon excited fluorescence (TPEF) in the pathologic spinal cord. It was compared qualitatively and quantitatively to Iba1 and CD68 immunohistochemical staining in two models: rat spinal cord injury and mouse encephalomyelitis. The extent of tissue damage was retrieved by coherent anti-Stokes Raman scattering (CARS) and second harmonic generation imaging. The pattern of CD68-positive cells representing postinjury activated microglia/macrophages was colocalized to the TPEF signal. Iba1-positive microglia were found in areas lacking any TPEF signal. In peripheral areas of inflammation, we found similar numbers of CD68-positive microglia/macrophages and TPEF-positive structures while the number of Iba1-positive cells was significantly higher. Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF. Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes.

No MeSH data available.


Related in: MedlinePlus

Quantitative analysis of TPEF-positive structures and the expression of microglial markers. The number of Iba1-, CD68-, and TPEF-positive structures was determined within the lesion and in peripheral tissue regions that displayed an inflammatory response. The bars represent mean ± SEM. ∗ indicates P < 0.05, one-way ANOVA followed by Tukey Multiple Comparison test.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4555451&req=5

fig3: Quantitative analysis of TPEF-positive structures and the expression of microglial markers. The number of Iba1-, CD68-, and TPEF-positive structures was determined within the lesion and in peripheral tissue regions that displayed an inflammatory response. The bars represent mean ± SEM. ∗ indicates P < 0.05, one-way ANOVA followed by Tukey Multiple Comparison test.

Mentions: The number of TPEF-, Iba1-, and CD68-positive structures was determined in different parts of all samples investigated. As the observed patterns were found to be comparable at both timepoints investigated, data of all samples was merged. The results are resumed in Figure 3. We found 3534 ± 397/mm−2 Iba1-positive cells at the lesion border. The number of CD68-positive cells and structures exhibiting endogenous TPEF was slightly lower in the same area, being 3111 ± 407/mm−2 and 3064 ± 310/mm−2, respectively.


Endogenous Two-Photon Excited Fluorescence Provides Label-Free Visualization of the Inflammatory Response in the Rodent Spinal Cord.

Uckermann O, Galli R, Beiermeister R, Sitoci-Ficici KH, Later R, Leipnitz E, Neuwirth A, Chavakis T, Koch E, Schackert G, Steiner G, Kirsch M - Biomed Res Int (2015)

Quantitative analysis of TPEF-positive structures and the expression of microglial markers. The number of Iba1-, CD68-, and TPEF-positive structures was determined within the lesion and in peripheral tissue regions that displayed an inflammatory response. The bars represent mean ± SEM. ∗ indicates P < 0.05, one-way ANOVA followed by Tukey Multiple Comparison test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4555451&req=5

fig3: Quantitative analysis of TPEF-positive structures and the expression of microglial markers. The number of Iba1-, CD68-, and TPEF-positive structures was determined within the lesion and in peripheral tissue regions that displayed an inflammatory response. The bars represent mean ± SEM. ∗ indicates P < 0.05, one-way ANOVA followed by Tukey Multiple Comparison test.
Mentions: The number of TPEF-, Iba1-, and CD68-positive structures was determined in different parts of all samples investigated. As the observed patterns were found to be comparable at both timepoints investigated, data of all samples was merged. The results are resumed in Figure 3. We found 3534 ± 397/mm−2 Iba1-positive cells at the lesion border. The number of CD68-positive cells and structures exhibiting endogenous TPEF was slightly lower in the same area, being 3111 ± 407/mm−2 and 3064 ± 310/mm−2, respectively.

Bottom Line: Iba1-positive microglia were found in areas lacking any TPEF signal.Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF.Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes.

View Article: PubMed Central - PubMed

Affiliation: Neurosurgery, Carl Gustav Carus University Hospital, TU Dresden, Fetscherstraße 74, 01307 Dresden, Germany.

ABSTRACT
Activation of CNS resident microglia and invasion of external macrophages plays a central role in spinal cord injuries and diseases. Multiphoton microscopy based on intrinsic tissue properties offers the possibility of label-free imaging and has the potential to be applied in vivo. In this work, we analyzed cellular structures displaying endogenous two-photon excited fluorescence (TPEF) in the pathologic spinal cord. It was compared qualitatively and quantitatively to Iba1 and CD68 immunohistochemical staining in two models: rat spinal cord injury and mouse encephalomyelitis. The extent of tissue damage was retrieved by coherent anti-Stokes Raman scattering (CARS) and second harmonic generation imaging. The pattern of CD68-positive cells representing postinjury activated microglia/macrophages was colocalized to the TPEF signal. Iba1-positive microglia were found in areas lacking any TPEF signal. In peripheral areas of inflammation, we found similar numbers of CD68-positive microglia/macrophages and TPEF-positive structures while the number of Iba1-positive cells was significantly higher. Therefore, we conclude that multiphoton imaging of unstained spinal cord tissue enables retrieving the extent of microglia activation by acquisition of endogenous TPEF. Future application of this technique in vivo will enable monitoring inflammatory responses of the nervous system allowing new insights into degenerative and regenerative processes.

No MeSH data available.


Related in: MedlinePlus