Simple Characterization of Cylindrical Diffuser Fibers With a Fluorescent Layer.

OBJECTIVES: A fast, simple, versatile, and reliable method to record light emission intensity profiles of cylindrical light diffusers (CDFs) in air and transparent liquids has been developed. METHODS: A fluorescent color glass filter (RG695) converts red light emitted by a cylindrical diffuser fiber into near-infrared light in an emission angle-independent manner. The red light was provided from a diode laser system at 635 nm. Near-infrared fluorescence from the RG695 was imaged with a camera. Images from this camera were processed to obtain emission intensity profiles. Cylindrical diffuser fiber profiles of four different manufacturers were compared. RESULTS: The proposed method provides angle-independent intensity profiles of cylindrical diffuser fibers with a single camera shot. It could be demonstrated that dependent on the underlying principle of how the diffuser fiber tips emit light, the emission profile can change significantly in media with different refractive indices. CONCLUSIONS: By converting the light emitted by a diffuser fiber tip into fluorescence light one can eliminate the dependence of the recorded profile on the emission angle from the diffusor. This approach allows for easily taking into account refraction-index (mis)matching by placing the equipment into a suitable liquid. The proposed measurement principle bears potential for quality assurance measurements of CDFs used for interstitial laser thermotherapy or photodynamic therapy.

Influence of vitamins and food on the fluorescence spectrum of human urine.

OBJECTIVES: Fluorescence spectroscopy of human urine is a method with the potential to gain importance as a diagnostic tool in the medical field, e.g., for measuring Coproporphyrin III (CPIII) as an indicator of cancer and acute types of porphyria. Food can change human urine's color, which could influence the urine fluorescence spectrum and the detection of CPIII in urine. To determine if there is a noticeable influence on the urine fluorescence spectrum or on the detection of CPIII in urine, 16 vitamin supplements, and three food items were tested. Such investigation may also prevent false interpretation of measured data. METHODS: Urine samples were collected before and after (overnight, ca. 8 h) intake of each test substance. Samples were investigated by fluorescence spectrum analysis. At excitation wavelengths from 300 to 500 nm and emission wavelengths from 400 to 700 nm excitation-emission-matrices were measured. Data obtained from urine before intake were compared to the data from overnight urine. Furthermore, the investigation of any interference with the CPIII concentration was performed at an excitation wavelength of 407 ± 3 nm and emission wavelengths of 490-800 nm. RESULTS: Only vitamin B2, but none of the other tested substances, showed noticeable influence on the urine fluorescence spectrum. None of the tested substances showed noticeable interference with the recovery rate of CPIII. CONCLUSIONS: The correct interpretation of measured data by fluorescence spectroscopy is possible with the exception if vitamin B2 supplementation was performed; thus, the consumption of vitamin B2 supplements before fluorescence testing of the patient's urine should be avoided and/or must be requested. CPIII concentrations could reliably be measured in all cases.

Interstitial photodynamic therapy for newly diagnosed glioblastoma.

PURPOSE: Innovative, efficient treatments are desperately needed for people with glioblastoma (GBM). METHODS: Sixteen patients (median age 65.8 years) with newly diagnosed, small-sized, not safely resectable supratentorial GBM underwent interstitial photodynamic therapy (iPDT) as upfront eradicating local therapy followed by standard chemoradiation. 5-aminolevulinic acid (5-ALA) induced protoporphyrin IX was used as the photosensitizer. The tumors were irradiated with light at 635 nm wavelength via stereotactically implanted cylindrical diffuser fibers. Outcome after iPDT was retrospectively compared with a positively-selected in-house patient cohort (n = 110) who underwent complete tumor resection followed by chemoradiation. RESULTS: Median progression-free survival (PFS) was 16.4 months, and median overall survival (OS) was 28.0 months. Seven patients (43.8%) experienced long-term PFS > 24 months. Median follow-up was 113.9 months for the survivors. Univariate regression revealed MGMT-promoter methylation but not age as a prognostic factor for both OS (p = 0.04 and p = 0.07) and PFS (p = 0.04 and p = 0.67). Permanent iPDT-associated morbidity was seen in one iPDT patient (6.3%). Patients treated with iPDT experienced superior PFS and OS compared to patients who underwent complete tumor removal (p < 0.01 and p = 0.01, respectively). The rate of long-term PFS was higher in iPDT-treated patients (43.8% vs. 8.9%, p < 0.01). CONCLUSION: iPDT is a feasible treatment concept and might be associated with long-term PFS in a subgroup of GBM patients, potentially via induction of so far unknown immunological tumor-controlling processes.

Non-Invasive Zinc Protoporphyrin Screening Offers Opportunities for Secondary Prevention of Iron Deficiency in Blood Donors.

Background: Frequent blood donors are at high risk of developing iron deficiency. Currently, there is no potent screening during blood donation to detect iron deficient erythropoiesis (IDE) before anemia develops and deferral from donation is inevitable. Study Design and Methods: In addition to capillary and venous hemoglobin, the iron status of 99 frequent blood donors was assessed by various venous blood parameters and zinc protoporphyrin IX (ZnPP). ZnPP was determined by high-performance liquid chromatography (HPLC) and a new prototype fiber-optic device was employed for non-invasive measurements of ZnPP through the blood collection tubing (NI-tubing) and on lip tissue (NI-lip). We aimed to evaluate the feasibility and diagnostic value of the NI-tubing measurement for early detection of severe iron deficiency in blood donors. Results: NI-tubing and HPLC reference measurements of ZnPP showed narrow limits of agreement of 12.2 µmol ZnPP/mol heme and very high correlation (Spearman's Rho = 0.938). Using a cutoff of 65 µmol ZnPP/mol heme, NI-tubing measurements (n = 93) identified 100% of donors with iron deficiency anemia (IDA) and an additional 38% of donors with IDE. Accordingly, NI-tubing measurements would allow detection and selective protection of particularly vulnerable donors. Conclusion: NI-tubing measurements are an accurate and simple method to implement ZnPP determination into the routine blood donation process. ZnPP was able to identify the majority of subjects with IDE and IDA and might therefore be a valuable tool to provide qualified information to donors about dietary measures and adjustments of the donation interval and thereby help to prevent IDA and hemoglobin deferral in the future.

Screening for iron deficiency in surgical patients based on noninvasive zinc protoporphyrin measurements.

BACKGROUND: Approximately every third surgical patient is anemic. The most common form, iron deficiency anemia, results from persisting iron-deficient erythropoiesis (IDE). Zinc protoporphyrin (ZnPP) is a promising parameter for diagnosing IDE, hitherto requiring blood drawing and laboratory workup. STUDY DESIGN AND METHODS: Noninvasive ZnPP (ZnPP-NI) measurements are compared to ZnPP reference determination of the ZnPP/heme ratio by high-performance liquid chromatography (ZnPP-HPLC) and the analytical performance in detecting IDE is evaluated against traditional iron status parameters (ferritin, transferrin saturation [TSAT], soluble transferrin receptor-ferritin index [sTfR-F], soluble transferrin receptor [sTfR]), likewise measured in blood. The study was conducted at the University Hospitals of Frankfurt and Zurich. RESULTS: Limits of agreement between ZnPP-NI and ZnPP-HPLC measurements for 584 cardiac and noncardiac surgical patients equaled 19.7 µmol/mol heme (95% confidence interval, 18.0-21.3; acceptance criteria, 23.2 µmol/mol heme; absolute bias, 0 µmol/mol heme). Analytical performance for detecting IDE (inferred from area under the curve receiver operating characteristics) of parameters measured in blood was: ZnPP-HPLC (0.95), sTfR (0.92), sTfR-F (0.89), TSAT (0.87), and ferritin (0.67). Noninvasively measured ZnPP-NI yielded results of 0.90. CONCLUSION: ZnPP-NI appears well suited for an initial IDE screening, informing on the state of erythropoiesis at the point of care without blood drawing and laboratory analysis. Comparison with a multiparameter IDE test revealed that ZnPP-NI values of 40 µmol/mol heme or less allows exclusion of IDE, whereas for 65 µmol/mol heme or greater, IDE is very likely if other causes of increased values are excluded. In these cases (77% of our patients) ZnPP-NI may suffice for a diagnosis, while values in between require analyses of additional iron status parameters.

Indocyanine green fluorescence imaging during partial adrenalectomy.

BACKGROUND: Indocyanine green (ICG) fluorescence imaging represents an emerging technology that facilitates the assessment of tissue vascularity, tissue distinction, and tumor localization during surgery. The aim of this study was to investigate the potential role of ICG imaging during laparoscopic partial adrenalectomy. METHODS: Indocyanine fluorescence imaging was carried out during laparoscopic partial adrenalectomy for bilateral pheochromocytoma and bilateral Cushing's syndrome. A first bolus of 5 mg ICG was applied intravenously upon exposure of the retroperitoneal plane to identify the adrenal borders. The fluorescence was visualized using a Storz® NIR/ICG endoscopic system. As the camera of this system detects NIR light as a blue signal, the well-vascularized adrenal tissue was expected to show a strong fluorescence in the blue color channel in contrast to the surrounding adipose tissue. Following partial adrenalectomy, a second bolus of 5 mg ICG was applied intravenously to evaluate the vascularity of the remaining adrenal tissue. RESULTS: We investigated six adrenal glands from three patients undergoing bilateral partial adrenalectomy. The indication for surgery was pheochromocytoma in two patients and Cushing's syndrome with bilateral adenomas in one patient. Regarding left adrenalectomies, ICG imaging was helpful in visualizing the adrenal borders and the adrenal vein. Further, it facilitated the identification of the hypofluorescent pheochromocytoma and to resect the entire tumor. On the right side, due to the more apparent anatomy, ICG imaging did not contribute to the conduct of the operation. Four adrenal remnants showed a strong vascularization and two remnants were only reasonably vascularized. CONCLUSION: ICG fluorescence may be helpful in guiding partial adrenalectomy and assessing the vascularity of remaining adrenal tissue.

Non-invasive measurement of erythrocyte zinc protoporphyrin in children.

BACKGROUND: Reliable screening for iron deficiency (ID) has required a blood sample and cost-intensive laboratory measurements. A novel method to non-invasively measure erythrocyte zinc protoporphyrin (ZnPP), an established marker for ID, is evaluated in children. METHODS: ZnPP was determined non-invasively by fluorescence measurements on the wet vermillion of the lower lip in 99 hospitalized children aged 9 months to 5 years. For comparison, conventional ID parameters and ZnPP were determined from blood samples. RESULTS: The non-invasively measured ZnPP values had limits of agreement (LoA) of 14 µmol ZnPP/mol heme (95% confidence interval: 9-20) compared to fluorescence measurements directly in blood. Repeated high-performance liquid chromatography reference determinations had comparable LoA of 14 µmol ZnPP/mol heme (9-17). Non-invasive ZnPP measurements had sensitivity and specificity of 67% (39-88%) and 97% (91-99%), and negative and positive predictive value of 94% (90-97%) and 80% (55-93%), for detecting ID as defined by the soluble transferrin receptor (sTfR). In groups with more severe ID as defined by serum ferritin and sTfR, higher ZnPP values were found, with the highest ZnPP values for the group with ID anemia. CONCLUSION: Non-invasive ZnPP measurements are reliably feasible in children. The simple and fast method has the potential to enable wide-spread screening for ID.

Parathyroid Autofluorescence-How Does It Affect Parathyroid and Thyroid Surgery? A 5 Year Experience.

Injury to parathyroid glands during thyroid and parathyroid surgery is common and postoperative hypoparathyroidism represents a serious complication. Parathyroid glands possess a unique autofluorescence in the near-infrared spectrum which could be used for their identification and protection at an early stage of the operation. In the present study parathyroid autofluorescence was visualized intraoperatively using a standard Storz laparoscopic near-infrared/indocyanine green (NIR/ICG) imaging system with minor modifications to the xenon light source (filtered to emit 690 nm to 790 nm light, less than 1% in the red and green above 470 nm and no blue light). During exposure to NIR light parathyroid tissue was expected to show autofluorescence at 820 nm, captured in the blue channel of the camera. Over a period of 5 years, we investigated 205 parathyroid glands from 117 patients. 179 (87.3%) glands were correctly identified by their autofluorescence. Surrounding structures such as thyroid, lymph nodes, muscle, or adipose tissue did not reveal substantial autofluorescence. We conclude that parathyroid glands can be identified by their unique autofluorescence at an early stage of the operation. This may help to preserve these fragile structures and their vascularization and lower the rate of postoperative hypocalcemia.

5-ALA in the management of malignant glioma.

BACKGROUND: Patients suffering from malignant gliomas have a poor prognosis. For the surgical treatment of these tumors, 5-aminolevulinic acid (5-ALA) has become a new standard. AIMS: This review intends to provide an overview over current status, significance, limitations, and future perspectives of 5-ALA based fluorescence guided surgery and photodynamic therapy for brain tumor patients. MATERIALS AND METHODS: From peer reviewed publications on the many aspects connected with this topic, those with potential clinical relevance were selected and put in the context of our own experience. RESULTS AND DISCUSSION: The high tumor selectivity of accumulation of fluorescent protoporphyrin IX (PpIX) after systemic administration of 5-ALA enables intra-operative fluorescence guidance, which is unimpaired by brainshift and does not require expensive equipment. The neurosurgical aim of complete resection of enhancing tumor can now more easily be achieved, which improves prognosis in these patients. Nevertheless, despite better surgery tumors will inevitably recur. In order to further prolong survival, the phototoxic properties of PpIX are presently being exploited in clinical trials of post-operative or interstitial photodynamic therapy (PDT). CONCLUSION: 5-ALA based fluorescence guidance and PDT offer an intriguing new option for the management of malignant gliomas. Lasers Surg. Med. 50:399-419, 2018. © 2018 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Intraoperative near-infrared autofluorescence imaging of parathyroid glands.

OBJECTIVE: To identify parathyroid glands intraoperatively by exposing their autofluorescence using near-infrared light. METHODS: Fluorescence imaging was carried out during minimally invasive and open parathyroid and thyroid surgery. After identification, the parathyroid glands as well as the surrounding tissue were exposed to near-infrared (NIR) light with a wavelength of 690-770 nm using a modified Karl Storz near-infrared/indocyanine green (NIR/ICG) endoscopic system. Parathyroid tissue was expected to show near-infrared autofluorescence, captured in the blue channel of the camera. Whenever possible the visual identification of parathyroid tissue was confirmed histologically. RESULTS: In preliminary investigations, using the original NIR/ICG endoscopic system we noticed considerable interference of light in the blue channel overlying the autofluorescence. Therefore, we modified the light source by interposing additional filters. In a second series, we investigated 35 parathyroid glands from 25 patients. Twenty-seven glands were identified correctly based on NIR autofluorescence. Regarding the extent of autofluorescence, there were no noticeable differences between parathyroid adenomas, hyperplasia and normal parathyroid glands. In contrast, thyroid tissue, lymph nodes and adipose tissue revealed no substantial autofluorescence. CONCLUSION: Parathyroid tissue is characterized by showing autofluorescence in the near-infrared spectrum. This effect can be used to distinguish parathyroid glands from other cervical tissue entities.

Photodynamic diagnosis with 5-aminolevulinic acid for intraoperative detection of peritoneal metastases of ovarian cancer: A feasibility and dose finding study.

OBJECTIVE: With a prospective feasibility study, we aimed to analyse the effect of different time points for application and dosage of preoperative oral 5-aminolevulinic acid administration for photodynamic diagnosis of peritoneal metastases in ovarian cancer patients. MATERIALS AND METHODS: In this prospective cohort study patients were randomly divided into three different groups. 5-Aminolevulinic acid was orally administered 3-14 hours before surgery using a dosage of 1 mg/kg, 4-9 hours using 10 mg/kg, and 9-16 hours using 10 mg/kg, respectively. Fluorescence was recorded intraoperatively using endoscopic equipment. The number and localization, of fluorescing nodules were documented. To analyze sensitivity and specificity samples from fluorescent and non-fluorescent tissues were evaluated histologically. Plasma protoporphyrin concentrations as well as any adverse events were assessed perioperatively. RESULTS: In total, 26 patients suspected for ovarian cancer underwent intraoperative photodynamic diagnosis with 5-aminolevulinic acid. Most of them suffered from advanced cancer, 72% from FIGO-Stage IIIc. No severe adverse events were observed. Orally applied 5-aminolevulinic acid with a dosage of 1 mg/kg revealed no detectable fluorescence. However, at a dosage of 10 mg/kg fluorescence of metastatic tissue was significantly stronger than of non-affected tissue. If administered 4-9 hours preoperatively best detection rates for peritoneal metastases were obtained resulting in a sensitivity of 75% and a specificity of 100%. CONCLUSIONS: Photodynamic diagnosis with 5-aminolevulinic acid leads to safe and specific fluorescence detection of peritoneal metastases. 5-Aminolevulinic acid should be used at a dosage of at least 10 mg/kg 4-9 hours preoperatively. Further phase I-II studies are recommended. Lasers Surg. Med. 49:169-176, 2017. © 2016 Wiley Periodicals, Inc.

Intraoperative assessment of laryngeal pathologies with optical coherence tomography integrated into a surgical microscope.

OBJECTIVE: Endoscopic examination followed by tissue biopsy is the gold standard in the evaluation of lesions of the upper aerodigestive tract. However, it can be difficult to distinguish between healthy mucosa, dysplasia, and invasive carcinoma. Optical coherence tomography (OCT) is a non-invasive technique which acquires high-resolution, cross-sectional images of tissue in vivo. Integrated into a surgical microscope, it allows the intraoperative evaluation of lesions simultaneously with microscopic visualization. STUDY DESIGN: In a prospective case series, we evaluated the use of OCT integrated into a surgical microscope during microlaryngoscopy to help differentiating various laryngeal pathologies. METHODS: 33 patients with laryngeal pathologies were examined with an OCT- microscope (OPMedT iOCT-camera, HS Hi-R 1000G-microscope, Haag-Streit Surgical GmbH, Wedel, Germany) during microlaryngoscopy. The suspected intraoperative diagnoses were compared to the histopathological reports of subsequent tissue biopsies. RESULTS: Hands-free non-contact OCT revealed high-resolution images of the larynx with a varying penetration depth of up to 1.2 mm and an average of 0.6 mm. Picture quality was variable. OCT showed disorders of horizontal tissue layering in dysplasias with a disruption of the basement membrane in carcinomas. When comparing the suspected diagnosis during OCT-supported microlaryngoscopy with histology, 79% of the laryngeal lesions could be correctly identified. Premalignant lesions were difficult to diagnose and falsely classified as carcinoma. CONCLUSION: OCT integrated into a surgical microscope seems to be a promising adjunct tool to discriminate pathologies of the upper aerodigestive tract intraoperatively. However, picture quality and penetration depth were variable. Although premalignant lesions were difficult to diagnose, the system proved overall helpful for the intraoperative discrimination of benign and malignant tumors. Further studies will be necessary to define its value in the future. Lasers Surg. Med. 49:490-497, 2017. © 2017 Wiley Periodicals, Inc.

Probe-based confocal laser endomicroscopy (pCLE): a preclinical investigation of the male genital tract.

The aim of this study was to assess the potential of probe-based confocal laser endomicroscopy (pCLE) as a new diagnostic imaging technique for the male genital tract. For this purpose, testes, epididymides, and vasa deferentia were obtained during transsexual surgery of healthy patients (n = 10, 26-52 years). Prior to this, testes of rats (n = 10, Sprague-Dawley) and mice (n = 8, wild-type) were examined. Ex vivo tissues were investigated by pCLE after topical fluorescence staining. Images and pCLE real-time video sequences were compared to images acquired by confocal laser scanning microscopy (CLSM); this allowed the identifying of corresponding microstructures. Interestingly, the seminiferous tubules of transsexual humans contained mainly spermatogonia due to long-term estrogen treatment, whereas the seminiferous tubules of the murine and rat spermatogenesis-related cell types were differentiated. Mosaicking improved the inspection potential by wide-angle views. Similarly, the microarchitecture of the epididymis and the vas deferens was successfully visualized in situ and on a cellular level by pCLE. In summary, pCLE allows for real-time identification of relevant microstructures responsible for spermatogenesis under ex vivo conditions. Additionally, pCLE enabled to localize vital spermatozoa in the testis thus opening up new ways to improve sperm retrieval rates during assisted reproduction. Both clinically relevant experiences hold promise to introduce this diagnostic method into a clinical study, and to investigate its potential as a clinical diagnostic procedure to expedite and improve the medical situation.

Intraoperative optical coherence tomography imaging to identify parathyroid glands.

OBJECTIVE: Optical coherence tomography (OCT) is a non-invasive high-resolution imaging technique that permits characterization of microarchitectural features in real time. Previous ex vivo studies have shown that the technique is capable of distinguishing between parathyroid tissue, thyroid tissue, lymph nodes, and adipose tissue. The purpose of this study was to evaluate the practicality of OCT during open and minimally invasive parathyroid and thyroid surgery. METHODS: During parathyroid and thyroid surgery, OCT images were generated from parathyroid glands, thyroid tissue, lymph nodes, and adipose tissue. The images were immediately assessed by the operating team using the previously defined criteria. Second, the OCT images were blinded with respect to their origin and analyzed by two investigators. Whenever possible the OCT findings were matched to the corresponding histology. RESULTS: A total of 227 OCT images from 27 patients undergoing open or minimally invasive thyroid or parathyroid surgery were analyzed. Parathyroid glands were correctly identified in 69.2%, thyroid tissue in 74.5%, lymph nodes in 37.5%, and adipose tissue in 69.2%. 43 OCT images (18.9%) could not be allocated to one of the tissue types (Table 2). Sensitivity and specificity in distinguishing parathyroid tissue from the other entities were 69% (63 true positive, 13 false negative findings, 15 images where an allocation was not possible) and 66%, respectively (71 true negative, 9 false positive, 28 images where an assessment was not possible). CONCLUSION: OCT is capable of distinguishing between parathyroid, thyroid, and adipose tissue. An accurate differentiation between parathyroid tissue and lymph nodes was not possible. The disappointing results compared to the previous ex vivo study are related to problems handling the endoscopic probe intraoperatively. However, further refinement of this new technology may lead to OCT systems with higher resolution and intraoperative probes that are easier to handle.

Effect of optical clearing agents on optical coherence tomography images of cervical epithelium.

Optical coherence tomography (OCT) can be used as an adjunct to colposcopy in order to detect precancerous and cancerous cervical lesions. Optical clearing agents (OCAs) temporarily reduce the optical scattering of biological tissues. The purpose of this study was to investigate their influence on OCT imaging. OCT images were taken from unsuspicious and suspicious areas of fresh conization specimens immediately after resection and 5, 10, and 20 min after application of dimethyl sulfoxide (DMSO) or polyethylene glycol (PEG). Corresponding histologies were obtained from all sites. The images taken 5, 10, and 20 min after application of OCA were compared to the initial images with respect to changes in brightness, contrast, and scanning depth using a standard nonparametric test of differences of proportions. Further, mean intensity backscattering curves were calculated from all OCT images in the histological groups CIN2, CIN3, inflammation, and normal epithelium. Mean difference profiles within each of these groups were determined, reflecting the mean differences between the condition before the application of OCA and the exposure times 5, 10, and 20 min, respectively. The null hypothesis was tested employing the Dicky-Fuller-test, Hotelings-test and run test. The visual analysis of 434 OCT images from 109 different sites of 24 conization specimens showed a statistically significant increase in brightness and contrast for normal and dysplastic epithelium after application of DMSO or PEG. Further, the analysis of mean intensity profiles suggests the existence of an increased backscattering intensity after application of DMSO or PEG. DMSO and PEG contribute substantially to optical clearing in cervical squamous epithelium and therefore influence OCT imaging in a positive way. With further refinement of the OCT technology, the observed changes may be beneficial in interpreting the tissue microstructure and identifying cervical intraepithelial neoplasia.

Effect of acetic acid on optical coherence tomography (OCT) images of cervical epithelium.

Optical coherence tomography (OCT) can be used as an adjunct to colposcopy in the identification of precancerous and cancerous cervical lesions. The purpose of this study was to investigate the effect of acetic acid on OCT imaging. OCT images were taken from unsuspicious and suspicious areas of fresh conization specimens immediately after resection and 3 and 10 min after application of 6 % acetic acid. A corresponding histology was obtained from all sites. The images taken 3 and 10 min after application of acetic acid were compared to the initial images with respect to changes in brightness, contrast, and scanning depth employing a standard nonparametric test of differences of proportions. Further, mean intensity backscattering curves were calculated from all OCT images in the histological groups CIN3, inflammation, or normal epithelium. Mean difference profiles within each of these groups were determined, reflecting the mean differences between the condition before application of acetic acid and the exposure times 3 and 10 min, respectively. According to the null hypothesis, the difference profiles do not differ from profiles fluctuating around zero in a stationary way, which implies that the profiles do not differ significantly from each other. The null hypothesis was tested employing the KPSS test. The visual analysis of 137 OCT images from 46 sites of 10 conization specimens revealed a statistically significant increase in brightness for all three groups and a statistically significant decrease in contrast for normal epithelium after 10 min. Further, an increase in scanning depth was noted for normal epithelium after 10 min and for CIN3 after 3 min. The analysis of mean intensity profiles showed an increased backscattering intensity after application of acetic acid. Acetic acid significantly affects the quality of OCT images. Overall brightness and scanning depth increase with the opposite effect regarding the image contrast. Whether the observed changes facilitate the distinction between dysplastic lesions in a clinical setting needs to be shown in further studies.

Comparison of an accelerated weighted fluorescence Monte Carlo simulation method with reference methods in multi-layered turbid media.

Monte Carlo (MC) simulations are frequently used to simulate the radial distribution of remitted fluorescence light from tissue surfaces upon pencil beam excitation to gather information about influences of different tissue parameters. Here, the "weighted direct emission method" (WDEM) is proposed, which uses a weighted MC simulation approach for both excitation and fluorescence photons, and is compared to four other methods in terms of accuracy and speed, and using a broad range of tissue-relevant optical parameters. The WDEM is 5.2× faster on average than a fixed weight MC approach while still preserving its accuracy. Additional gain of speed can be achieved by implementing it on graphics processing units.

Protoporphyrin IX fluorescence and photobleaching during interstitial photodynamic therapy of malignant gliomas for early treatment prognosis.

BACKGROUND AND OBJECTIVE: Interstitial photodynamic therapy (iPDT) of non-resectable recurrent glioblastoma using 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) has shown a promising outcome. It remained unclear, however, to what extent inter- and intra-tumoural differences of PpIX concentrations influence the efficacy of iPDT. In the current pilot study, we analysed PpIX concentrations quantitatively and assessed PpIX induced fluorescence and photobleaching intraoperatively. MATERIALS AND METHODS: Five patients harbouring non-resectable glioblastomas were included. ALA (20 or 30 mg/kg body weight) was given 5-8 hours before treatment. Stereotactic biopsies were taken throughout the tumour volume for both histological analysis and determination of PpIX concentrations, which were measured by chemical extraction. Cylindrical light diffusors were stereotactically implanted. Prior to and after irradiation, fluorescence measurements were performed. Outcome measurement was based on clinical and neuro-radiological follow up. RESULTS: In three patients, a strong PpIX fluorescence was seen before treatment, which was completely photobleached after iPDT. High concentrations of PpIX could be detected in viable tumour parts of these patients (mean PpIX uptake per tumour: 1.4-3.0 µM). In the other two patients, however, no or only low PpIX uptake (0-0.6 µM) could be detected. The patients with strong PpIX uptake showed treatment response and long-term clinical stabilisation (no progression in 29, 30 and 36 months), early treatment failure was seen in the remaining two patients (death after 3 and 9 months). CONCLUSIONS: Intra-tumoural PpIX concentrations exhibited pronounced inter- and intra-tumoural variations in glioblastoma, which are directly linked to variable degrees of fluorescence intensity. High intra-tumoural PpIX concentrations with strong fluorescence intensity and complete photobleaching after iPDT seem to be associated with favourable outcome. Real-time monitoring of PpIX fluorescence intensity and photobleaching turned out to be feasible and safe and might be employed for early treatment prognosis of iPDT.

Optical coherence tomography as a method to identify parathyroid glands.

BACKGROUND AND OBJECTIVE: The identification of parathyroid glands can be a major problem in parathyroid surgery. The purpose of this study was to evaluate the feasibility of optical coherence tomography (OCT) in distinguishing between parathyroid tissue, thyroid tissue, lymph nodes, and adipose tissue. METHODS: Ex vivo OCT images as well as histological sections were generated from parathyroid glands, thyroid tissue, lymph nodes and fat in order to define significant morphologic differences between these entities. As a second step all OCT images were separately evaluated by two blinded investigators and later compared to the corresponding histology. Sensitivity and specificity of OCT in distinguishing between the different tissues were determined. To assess the interobserver agreement, κ coefficients were calculated from the ratings of each investigator for each OCT image seen. RESULTS: A total of 320 OCT images from 32 patients undergoing thyroid surgery, parathyroidectomy or lymphadenectomy were compared with the corresponding histology. The sensitivity and specificity in distinguishing parathyroid tissue from the other entities was 84% (second investigator: 82%) and 94% (93%) respectively. Unweighted κ using four diagnostic categories was 0.97 (95% CI, 0.94-0.99) showing substantial agreement between both investigators. CONCLUSION: OCT is highly sensitive in distinguishing between parathyroid tissue, thyroid tissue, lymph nodes and adipose tissue. These ex vivo results should be confirmed by using OCT imaging intraoperatively.