Multiphoton excitation fluorescence microscopy of 5‐aminolevulinic acid induced fluorescence in experimental gliomas

Kantelhardt, Sven R.; Diddens, Heike; Leppert, Jan; Rohde, Veit; Hüttmann, Gereon; Giese, Alf

doi:10.1002/lsm.20623

Cited by 41 publications

(49 citation statements)

References 34 publications

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“…Yet the spectroscopy technique still needs to be validated in clinical routine. 6,10,23 Our data suggest a higher detection rate for tumor and infiltration zone in 5-ALA as opposed to iMRI. In Gd-DTPA-enhanced iMRI a high number of tumor-positive areas are missed.…”

Section: 29mentioning

confidence: 62%

Tumor detection with 5-aminolevulinic acid fluorescence and Gd-DTPA–enhanced intraoperative MRI at the border of contrast-enhancing lesions: a prospective study based on histopathological assessment

Coburger¹,

Engelke²,

Scheuerle

et al. 2014

FOC

125

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Object High-grade gliomas (HGGs) and metastasis (MET) are the most common intracranial lesions in neurosurgical routine. Both of them show an invasive growth pattern extending into neural tissue beyond the margins of contrast enhancement on MRI. These “undetected” areas might be the origin of early tumor recurrence. The aim of the present study was to evaluate whether 5-aminolevulinic acid (5-ALA) fluorescence provides an additional benefit in detection of invasive tumor compared with intraoperative MRI (iMRI). Methods The authors prospectively enrolled 45 patients harboring contrast-enhancing lesions, in whom gross-total resection was intended. All patients had surgery in which iMRI and 5-ALA–guided resection were used following a specific protocol. First, a typical white light tumor resection was performed. Then, spatial location of residual fluorescence was marked. After that, an iMRI was performed and residual uptake of contrast was marked. Navigated biopsy samples were taken from all marked areas and from additional sites according to the surgeon's judgment. Cross tables and receiver operating characteristic curves were calculated, assessing performance of the imaging methods for tumor detection alone and for combined detection of infiltration zone and solid tumor (pathological tissue). Also, correlations of histopathological findings with imaging results were tested using Spearman rho. Results Thirty-four patients with HGGs and 11 with METs were enrolled. Three patients harboring a MET showed no 5-ALA enhancement and were excluded; 127 histopathological samples were harvested in the remaining patients. In HGG, sensitivity for tumor detection was significantly higher (p < 0.001) in 5-ALA (0.85) than in iMRI (0.41). Specificity was significantly lower (p < 0.001) in 5-ALA (0.43) than in iMRI (0.70). For detection of pathological tissue, 5-ALA significantly exceeded iMRI in specificity (0.80 vs 0.60) and sensitivity (0.91 vs 0.66) (p < 0.001). Imaging results of iMRI and 5-ALA did not correlate significantly; only 5-ALA showed a significant correlation with final histopathological diagnosis of the specimen and with typical histopathological features of HGGs. In METs, sensitivity and specificity for tumor detection were equal in 5-ALA and iMRI. Both techniques showed high values for sensitivity (0.75) and specificity (0.80). The odds ratio for detection of tumor tissue was 12 for both techniques. Concerning pathological tissue, no statistically significant difference was found either. Imaging results of iMRI and 5-ALA correlated significantly (p < 0.022), as with final histopathological diagnosis in METs. Conclusions In METs, due to the rate of nonenhancing lesions, the authors found no additional benefit of 5-ALA compared with iMRI. In HGG, imaging results of 5-ALA and iMRI are significantly different at the border zone; 5-ALA has a higher sensitivity and a lower specificity for tumor detection than Gd-DTPA–enhanced iMRI. For detection of infiltrating tumor at the border of the resection cavity, 5-ALA is superior to Gd-DTPA–enhanced iMRI concerning both sensitivity and specificity. Thus, use of 5-ALA in addition to iMRI might be beneficial to maximize extent of resection. Clinical synergistic effects will be evaluated in a prospective randomized trial.

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Section: 29mentioning

confidence: 62%

Tumor detection with 5-aminolevulinic acid fluorescence and Gd-DTPA–enhanced intraoperative MRI at the border of contrast-enhancing lesions: a prospective study based on histopathological assessment

Coburger¹,

Engelke²,

Scheuerle

et al. 2014

FOC

125

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“…Cicchi et al [102] demonstrated enhanced tumor contrast and depth of imaging by using MAL-induced PpIX as the contrast agent, and two-photon excitation (TPE) in combination with FLIM as the imaging technique. A similar study by Kantelhardt et al [103] to identify gliomas achieved high contrast images of the glioma architecture that were easily discriminated from normal brain tissue, by detecting ALA-induced PpIX fluorescence with multiphoton microscopy. A few other studies report the use of multiphoton excitation for PpIX detection [104], but offer no detailed spectral information.…”

Section: Approaches For Measuring Ppix Beneath the Surface Of The Skinmentioning

confidence: 94%

Techniques for fluorescence detection of protoporphyrin IX in skin cancers associated with photodynamic therapy

Rollakanti

Kanick

Davis

et al. 2013

Photonics &Amp; Lasers in Medicine

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Photodynamic therapy (PDT) is a treatment modality that uses a specific photosensitizing agent, molecular oxygen, and light of a particular wavelength to kill cells targeted by the therapy. Topically administered aminolevulinic acid (ALA) is widely used to effectively treat cancerous and precancerous skin lesions, resulting in targeted tissue damage and little to no scarring. The targeting aspect of the treatment arises from the fact that ALA is preferentially converted into protoporphyrin IX (PpIX) in neoplastic cells. To monitor the amount of PpIX in tissues, techniques have been developed to measure PpIX-specific fluorescence, which provides information useful for monitoring the abundance and location of the photosensitizer before and during the illumination phase of PDT. This review summarizes the current state of these fluorescence detection techniques. Non-invasive devices are available for point measurements, or for wide-field optical imaging, to enable monitoring of PpIX in superficial tissues. To gain access to information at greater tissue depths, multi-modal techniques are being developed which combine fluorescent measurements with ultrasound or optical coherence tomography, or with microscopic techniques such as confocal or multiphoton approaches. The tools available at present, and newer devices under development, offer the promise of better enabling clinicians to inform and guide PDT treatment planning, thereby optimizing therapeutic outcomes for patients.

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“…This suggests that the photochemical analysis may offer means for specific detection of residual tumour in biopsy specimens, which may increase the diagnostic value of this technique. We have further demonstrated that the specificity of tumour cell detection by multi-photon microscopy may be significantly increased by detection of 5-ALA induced PpIX fluorescence in experimental gliomas [7]. The fluorescence lifetime analysis of induced PpIX fluorescence by multi-photon microscopy demonstrated that PpIX fluorescence was significantly prolonged compared to fluorescence of endogenous fluorophores.…”

Section: Discussionmentioning

confidence: 74%

Multi-photon excitation fluorescence microscopy of brain-tumour tissue and analysis of cell density

Kantelhardt

Leppert

Kantelhardt

et al. 2009

Acta Neurochir (Wien)

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Background Recent studies have highlighted the importance of the complete resection of a brain tumour but the task often remains a challenge for the neurosurgeon. New technologies which add objective information beyond visualisation provided by the traditional operating microscope are required. In this study, we have analysed the cellular density of the tumour/brain interface using three dimensional multi-photon microscopy intensity-images of experimental gliomas and human brain-tumour biopsy samples. Methods The density of cellular nuclei was determined in specimens of experimental gliomas in a mouse model and human brain tumour biopsies by analysis of optical tissue sections. Three dimensional multi-photon microscopy image stacks were compared to serial H&E stained sections of conventional histolopathology. Findings Both techniques consistently showed a good correlation of cell density values in solid tumour tissue of experimental gliomas versus adjacent brain. The multiphoton microscopy analysis of human biopsy specimens showed that optical analysis of native tissue provided information on the cellular density. Conclusions Multi-photon microscopy is an efficient and rapid tool for the study of brain and brain tumour tissue. Multi-photon microscopy allows the detection of individual tumour cells and tumour cell clusters in native tissue biopsies and may therefore provide a tool in the identification of highly cellular lesions during the resection of brain tumours.

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Multiphoton excitation fluorescence microscopy of 5‐aminolevulinic acid induced fluorescence in experimental gliomas

Abstract: Multiphoton microscopy of 5-ALA induced PpIX fluorescence in brain tumor tissue conceptually provides a high resolution diagnostic tool, which in addition to structural information may also provide photochemical/functional information.

Cited by 41 publications

References 34 publications

Tumor detection with 5-aminolevulinic acid fluorescence and Gd-DTPA–enhanced intraoperative MRI at the border of contrast-enhancing lesions: a prospective study based on histopathological assessment

Tumor detection with 5-aminolevulinic acid fluorescence and Gd-DTPA–enhanced intraoperative MRI at the border of contrast-enhancing lesions: a prospective study based on histopathological assessment

Techniques for fluorescence detection of protoporphyrin IX in skin cancers associated with photodynamic therapy

Multi-photon excitation fluorescence microscopy of brain-tumour tissue and analysis of cell density

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