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Confocal laser endomicroscopy for diagnosis and histomorphologic imaging of brain tumors in vivo.

Foersch S, Heimann A, Ayyad A, Spoden GA, Florin L, Mpoukouvalas K, Kiesslich R, Kempski O, Goetz M, Charalampaki P - PLoS ONE (2012)

Bottom Line: CLE discrimination of neoplastic from healthy brain tissue was easy to perform based on tissue and cellular architecture and resemblance with histopathology was excellent.Confocal laser endomicroscopy allows immediate in vivo imaging of normal and neoplastic brain tissue at high resolution.The technology might be transferred to scientific and clinical application in neurosurgery and neuropathology.

View Article: PubMed Central - PubMed

Affiliation: Medical Clinic I, University Medical Center, Mainz, Germany.

ABSTRACT
Early detection and evaluation of brain tumors during surgery is crucial for accurate resection. Currently cryosections during surgery are regularly performed. Confocal laser endomicroscopy (CLE) is a novel technique permitting in vivo histologic imaging with miniaturized endoscopic probes at excellent resolution. Aim of the current study was to evaluate CLE for in vivo diagnosis in different types and models of intracranial neoplasia. In vivo histomorphology of healthy brains and two different C6 glioma cell line allografts was evaluated in rats. One cell line expressed EYFP, the other cell line was used for staining with fluorescent dyes (fluorescein, acriflavine, FITC-dextran and Indocyanine green). To evaluate future application in patients, fresh surgical resection specimen of human intracranial tumors (n = 15) were examined (glioblastoma multiforme, meningioma, craniopharyngioma, acoustic neurinoma, brain metastasis, medulloblastoma, epidermoid tumor). Healthy brain tissue adjacent to the samples served as control. CLE yielded high-quality histomorphology of normal brain tissue and tumors. Different fluorescent agents revealed distinct aspects of tissue and cell structure (nuclear pattern, axonal pathways, hemorrhages). CLE discrimination of neoplastic from healthy brain tissue was easy to perform based on tissue and cellular architecture and resemblance with histopathology was excellent. Confocal laser endomicroscopy allows immediate in vivo imaging of normal and neoplastic brain tissue at high resolution. The technology might be transferred to scientific and clinical application in neurosurgery and neuropathology. It may become helpful to screen for tumor free margins and to improve the surgical resection of malignant brain tumors, and opens the door to in vivo molecular imaging of tumors and other neurologic disorders.

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Related in: MedlinePlus

Healthy cerebellum of a Wistar rat.A - In vivo CLE enabled to image the histology of different parts of the brain, e. g. the cerebellum. After topical application of acriflavine, the characteristic layers of can be observed and specific Purkinje cells (asterisk) with their dendritic tree (triangle) are stained. B - The resemblance to ex vivo histopathologic staining is substantial.
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pone-0041760-g002: Healthy cerebellum of a Wistar rat.A - In vivo CLE enabled to image the histology of different parts of the brain, e. g. the cerebellum. After topical application of acriflavine, the characteristic layers of can be observed and specific Purkinje cells (asterisk) with their dendritic tree (triangle) are stained. B - The resemblance to ex vivo histopathologic staining is substantial.

Mentions: In n = 11 allograft bearing rats in vivo confocal endomicroscopic imaging was performed (Table 2). In n = 3 animals, intravenous application of fluorescein resulted in a strong staining to observe overall brain and tumor morphology. First, the structure of small vessels and capillaries could be seen, after several minutes fluorescein diffused into both healthy brain and tumor tissue. Tissue staining could be observed faster and more intensively in malignant tissue compared to normal brain tissue due to capillary leakage via the incomplete blood-brain-barrier (BBB). Depending on the location of the confocal imaging window, healthy brain tissue showed a low number of prominent triangular cells, likely mimicking neurons. Smaller cells – potentially of glial origin - could also be observed and axon pathways, as well as medullar structures could be seen in deeper areas of the brain. CLE also enabled in vivo imaging of the cerebellum, where characteristic stratification into molecular layer, Purkinje layer, nuclear layer and medullar layer could be observed. Especially Purkinje cells could be seen clearly with their dendritic branches, large cell somas and tree like shapes (Figure 2). In tumor tissue strong fluorescence depicted massive extravasation and cell rich stroma could be displayed. Small intratumoral hemorrhages are represented by erythrocytes flowing into the interstitial space (Figure 3).


Confocal laser endomicroscopy for diagnosis and histomorphologic imaging of brain tumors in vivo.

Foersch S, Heimann A, Ayyad A, Spoden GA, Florin L, Mpoukouvalas K, Kiesslich R, Kempski O, Goetz M, Charalampaki P - PLoS ONE (2012)

Healthy cerebellum of a Wistar rat.A - In vivo CLE enabled to image the histology of different parts of the brain, e. g. the cerebellum. After topical application of acriflavine, the characteristic layers of can be observed and specific Purkinje cells (asterisk) with their dendritic tree (triangle) are stained. B - The resemblance to ex vivo histopathologic staining is substantial.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3404071&req=5

pone-0041760-g002: Healthy cerebellum of a Wistar rat.A - In vivo CLE enabled to image the histology of different parts of the brain, e. g. the cerebellum. After topical application of acriflavine, the characteristic layers of can be observed and specific Purkinje cells (asterisk) with their dendritic tree (triangle) are stained. B - The resemblance to ex vivo histopathologic staining is substantial.
Mentions: In n = 11 allograft bearing rats in vivo confocal endomicroscopic imaging was performed (Table 2). In n = 3 animals, intravenous application of fluorescein resulted in a strong staining to observe overall brain and tumor morphology. First, the structure of small vessels and capillaries could be seen, after several minutes fluorescein diffused into both healthy brain and tumor tissue. Tissue staining could be observed faster and more intensively in malignant tissue compared to normal brain tissue due to capillary leakage via the incomplete blood-brain-barrier (BBB). Depending on the location of the confocal imaging window, healthy brain tissue showed a low number of prominent triangular cells, likely mimicking neurons. Smaller cells – potentially of glial origin - could also be observed and axon pathways, as well as medullar structures could be seen in deeper areas of the brain. CLE also enabled in vivo imaging of the cerebellum, where characteristic stratification into molecular layer, Purkinje layer, nuclear layer and medullar layer could be observed. Especially Purkinje cells could be seen clearly with their dendritic branches, large cell somas and tree like shapes (Figure 2). In tumor tissue strong fluorescence depicted massive extravasation and cell rich stroma could be displayed. Small intratumoral hemorrhages are represented by erythrocytes flowing into the interstitial space (Figure 3).

Bottom Line: CLE discrimination of neoplastic from healthy brain tissue was easy to perform based on tissue and cellular architecture and resemblance with histopathology was excellent.Confocal laser endomicroscopy allows immediate in vivo imaging of normal and neoplastic brain tissue at high resolution.The technology might be transferred to scientific and clinical application in neurosurgery and neuropathology.

View Article: PubMed Central - PubMed

Affiliation: Medical Clinic I, University Medical Center, Mainz, Germany.

ABSTRACT
Early detection and evaluation of brain tumors during surgery is crucial for accurate resection. Currently cryosections during surgery are regularly performed. Confocal laser endomicroscopy (CLE) is a novel technique permitting in vivo histologic imaging with miniaturized endoscopic probes at excellent resolution. Aim of the current study was to evaluate CLE for in vivo diagnosis in different types and models of intracranial neoplasia. In vivo histomorphology of healthy brains and two different C6 glioma cell line allografts was evaluated in rats. One cell line expressed EYFP, the other cell line was used for staining with fluorescent dyes (fluorescein, acriflavine, FITC-dextran and Indocyanine green). To evaluate future application in patients, fresh surgical resection specimen of human intracranial tumors (n = 15) were examined (glioblastoma multiforme, meningioma, craniopharyngioma, acoustic neurinoma, brain metastasis, medulloblastoma, epidermoid tumor). Healthy brain tissue adjacent to the samples served as control. CLE yielded high-quality histomorphology of normal brain tissue and tumors. Different fluorescent agents revealed distinct aspects of tissue and cell structure (nuclear pattern, axonal pathways, hemorrhages). CLE discrimination of neoplastic from healthy brain tissue was easy to perform based on tissue and cellular architecture and resemblance with histopathology was excellent. Confocal laser endomicroscopy allows immediate in vivo imaging of normal and neoplastic brain tissue at high resolution. The technology might be transferred to scientific and clinical application in neurosurgery and neuropathology. It may become helpful to screen for tumor free margins and to improve the surgical resection of malignant brain tumors, and opens the door to in vivo molecular imaging of tumors and other neurologic disorders.

Show MeSH
Related in: MedlinePlus