scholarly journals BIMG-03. MOLECULAR IMAGING OF GLUCOSE METABOLISM FOR INTRAOPERATIVE FLUORESCENCE GUIDANCE DURING GLIOMA SURGERY

2021 ◽  
Vol 3 (Supplement_1) ◽  
pp. i1-i1
Author(s):  
Evgenii Belykh ◽  
Jubran Jubran ◽  
Laeth George ◽  
Liudmila Bardonova ◽  
Deborah Healey ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Brain Parenchyma ◽  
Confocal Laser ◽  
Normal Brain ◽  
Wide Field ◽  
Operating Microscope ◽  
Microscopy Imaging ◽  
Tumor Margins ◽  
Background Contrast ◽  
Intraoperative Fluorescence

Abstract PURPOSE This study evaluated the utility of using molecular imaging of fluorescent glucose analog 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) as a discriminatory marker for intraoperative tumor border identification in a mouse glioma model. PROCEDURES 2-NBDG and were assessed in GL261 and U251 orthotopic tumor bearing mice. Intraoperative fluorescence of 2-NBDG administered topical and intravenous in normal and tumor regions was assessed with operating microscope, handheld confocal laser scanning endomicroscope (CLE) and benchtop confocal laser scanning microscope (LSM). Additionally, 2-NBDG fluorescence in tumors was compared to 5-aminolevulinic acid-induced protoporphyrin IX fluorescence. RESULTS Intravenously administered 2-NBDG was detectable in brain tumor and absent in contralateral normal brain parenchyma on wide field operating microscopy imaging. Intraoperative and benchtop CLE showed preferential 2-NBDG accumulation in the cytoplasm of glioma cells (tumor-background ratio of 2.76±0.43). Topically administered 2-NBDG did not create a sufficient tumor-background contrast for white field operating microscopy imaging, or under benchtop LSM (tumor-background ratio 1.42 ± 0.72). However, topical 2-NBDG did create sufficient contrast to evaluate cellular tissue architecture and differentiate tumor cells from normal brain parenchyma. PpIX imaging resulted in a more specific delineation of gross tumor margins than IV or topical 2-NBDG, and a significantly higher tumor-normal brain fluorescence intensity ratio. CONCLUSION After intravenous administration, 2-NBDG selectively accumulated in the experimental brain tumors and provided bright contrast under wide field fluorescence imaging with a clinical grade operating microscope. Topical 2-NBDG was able to create a sufficient contrast to differentiate tumor from normal brain cells based on visualization of cellular architecture with CLE. 5-ALA demonstrated superior specificity in outlining tumor margins and significantly higher tumor-background contrast. Given its non-toxicity, using 2-NBDG as a topical molecular marker for noninvasive in vivo intraoperative microscopy is encouraging, and warrants further clinical evaluation.

scholarly journals Use of electromagnetic stimulation on an Enterococcus faecalis biofilm on root canal treated teeth in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Beatriz H. D. Panariello ◽  
Justin K. Kindler ◽  
Kenneth J. Spolnik ◽  
Ygal Ehrlich ◽  
George J. Eckert ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Confocal Imaging ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Microscopy Imaging ◽  
Electromagnetic Stimulation ◽  
Root Canal Disinfection

AbstractRoot canal disinfection is of utmost importance in the success of the treatment, thus, a novel method for achieving root canal disinfection by electromagnetic waves, creating a synergistic reaction via electric and thermal energy, was created. To study electromagnetic stimulation (EMS) for the disinfection of root canal in vitro, single rooted teeth were instrumented with a 45.05 Wave One Gold reciprocating file. Specimens were sterilized and inoculated with Enterococcus faecalis ATCC 29,212, which grew for 15 days to form an established biofilm. Samples were treated with 6% sodium hypochlorite (NaOCl), 1.5% NaOCl 1.5% NaOCl with EMS, 0.9% saline with EMS or 0.9% saline. After treatments, the colony forming units (CFU) was determined. Data was analyzed by Wilcoxon Rank Sums Test (α = 0.05). One sample per group was scored and split for confocal laser scanning microscopy imaging. There was a significant effect with the use of NaOCl with or without EMS versus 0.9% saline with or without EMS (p = 0.012 and 0.003, respectively). CFUs were lower when using 0.9% saline with EMS versus 0.9% saline alone (p = 0.002). Confocal imaging confirmed CFU findings. EMS with saline has an antibiofilm effect against E. faecalis and can potentially be applied for endodontic disinfection.

scholarly journals Silencing cuticular pigmentation genes enables RNA FISH in intact chemosensory appendagess

2018 ◽  
Author(s):  
Stefan Pentzold ◽  
Veit Grabe ◽  
Andrei Ogonkov ◽  
Lydia Schmidt ◽  
Wilhelm Boland ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Microscopy Imaging ◽  
Leaf Beetles ◽  
Antisense Mrna ◽  
Rna Fish ◽  
Whole Mount ◽  
Light Absorbance ◽  
Odorant Binding

AbstractOptical imaging of gene expression by RNA-fluorescent in situ hybridisation (FISH) in whole-mount sensory appendages of insects is often impeded by their highly pigmented cuticle. Since most chemical bleaching agents are incompatible with imaging fluorescent-labelled nucleotides, we developed a RNA interference-based method for clearing cuticular pigmentation that allows imaging of fluorescent mRNA in whole-mount appendages of insects. Silencing key genes of the tyrosine-derived pigmentation pathway by injecting dsRNA of laccase2 or tyrosine hydroxylase in two leaf beetles species (Chrysomela populi, Phaedon cochleariae) resulted in clearance of the highly pigmented cuticle and in significant decreased light absorbance. Intact chemosensory appendages (palps, antennae and legs) from RNAi-cleared individuals were used to image expression and spatial distribution of antisense mRNA of two chemosensory genes (gustatory receptor, odorant-binding protein) via RNA FISH and confocal laser scanning microscopy. Imaging of these genes did neither work for RNAi-controls (dsGfp) due to retained pigmentation, nor for FISH-controls using sense mRNA. Furthermore, we show that several chemical bleaching agents are not feasible with FISH, either due to significant degradation of polynucleotides, lack of clearing efficacy or long incubation times. Overall, silencing pigmentation genes is a significant improvement over bleaching agents allowing fluorescence imaging in whole-mount appendages and organs.

scholarly journals Redosing of Fluorescein Sodium Improves Image Interpretation During Intraoperative Ex Vivo Confocal Laser Endomicroscopy of Brain Tumors

2021 ◽  
Vol 11 ◽  
Author(s):  
Irakliy Abramov ◽  
Alexander B. Dru ◽  
Evgenii Belykh ◽  
Marian T. Park ◽  
Liudmila Bardonova ◽  
...  
Keyword(s):  
Single Dose ◽  
Image Quality ◽  
Ex Vivo ◽  
Confocal Laser Endomicroscopy ◽  
Confocal Laser ◽  
Wide Field ◽  
Fluorescein Sodium ◽  
Operating Microscope ◽  
Comparison Groups ◽  
Dose Imaging

BackgroundFluorescein sodium (FNa) is a fluorescence agent used with a wide-field operating microscope for intraoperative guidance and with confocal laser endomicroscopy (CLE) to evaluate brain tissue. Susceptibility of FNa to degradation over time may affect CLE image quality during prolonged surgeries. This study describes improved characteristics of CLE images after intraoperative redosing with FNa.MethodsA retrospective analysis was performed using CLE images obtained ex vivo from samples obtained during tumor resections with FNa-based fluorescence guidance with a wide-field operating microscope. The comparison groups included CLE images acquired after FNa redosing (redose imaging group), images from the same patients acquired after the initial FNa dose (initial-dose imaging group), and images from patients in whom redosing was not used (single-dose imaging group). A detailed assessment of image quality and interpretation regarding different FNa dosage and timing of imaging after FNa administration was conducted for all comparison groups.ResultsThe brightest and most contrasting images were observed in the redose group compared to the initial-dose and single-dose groups (P<0.001). The decay of FNa signal negatively correlated with brightness (rho = -0.52, P<0.001) and contrast (rho = -0.57, P<0.001). Different doses of FNa did not significantly affect the brightness (P=0.15) or contrast (P=0.09) in CLE images. As the mean timing of imaging increased, the percentage of accurately diagnosed images decreased (P=0.03).ConclusionsThe decay of the FNa signal is directly associated with image brightness and contrast. The qualitative interpretation scores of images were highest for the FNa redose imaging group. Redosing with FNa to improve the utility of CLE imaging should be considered a safe and beneficial strategy during prolonged surgeries.

Automated biofilm morphology quantification from confocal laser scanning microscopy imaging

2003 ◽  
Vol 47 (5) ◽  
pp. 31-37 ◽  
Author(s):  
J.B. Xavier ◽  
D.C. White ◽  
J.S. Almeida
Keyword(s):  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser ◽  
Microscopy Imaging ◽  
Biofilm Structure ◽  
User Friendly ◽  
Processing Steps

In spite of the immediate visual appeal of confocal laser scanning microscopy images, the extraction of accurate reconstitutions of biofilm morphology requires a lengthy and computational intensive succession of processing steps. However, once performed, it provides ample reward by enabling the quantitative study of biofilm structure. A software suite of image processing tools for full automation of biofilm morphology quantification was developed by integrating preprocessing, segmentation and morphology quantification operations. This software toolbox was implemented in a web server and a user friendly interface was developed to facilitate image submission, storage and sharing, its access being unrestricted for scientific applications. The image bioinformatics tool which results from the integration of the processing operations can be accessed at http://www.itqb.unl.pt:1111/clsmip/. Its use is described in this paper and is illustrated with an example of processing of experimental data describing the growth of a mixed species denitrifying biofilm.

The furfuryl alcohol (FA) resin distribution in the furfurylated bamboo

Holzforschung ◽  
2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Wanju Li ◽  
Minghui Liu ◽  
Hankun Wang ◽  
Hongbo Zhai ◽  
Yan Yu
Keyword(s):  
Furfuryl Alcohol ◽  
Laser Scanning ◽  
Parenchymal Cell ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Sem Images ◽  
Microscopy Imaging ◽  
Resin Impregnation ◽  
Specific Distribution ◽  
Ft Ir

AbstractIt is well known the properties of resin impregnation wood is significantly influenced by the specific distribution pattern of resin in the modified wood. In this work, bamboo was furfurylated with an improved process. In addition to testing and evaluating its main physical, mechanical and durable properties, it was explored how the furfuryl alcohol (FA) resin is distributed in the furfurylated bamboo. To achieve this goal, scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), nanoindentation, and imaging Fourier transform infrared microscopy (imaging FT-IR) were applied. SEM images demonstrates FA resin is mainly located in the parenchymal cell cavity of bamboo, however the existence of FA resin in the small cavities of the bamboo fibers is also observed by CLSM. In addition, the result of nanoindentation and imaging FT-IR both indicates that FA can penetrate and polymerize within the cell wall of the bamboo fiber. It is then concluded the heterogeneous but multi-scale distribution of FA resin should be responsible for the significant improvement of furfurylated bamboo in both dimensional stability and biological durability.

Assessment of three-dimensional biofilm models through direct comparison with confocal microscopy imaging

2004 ◽  
Vol 49 (11-12) ◽  
pp. 177-185 ◽  
Author(s):  
J.B. Xavier ◽  
C. Picioreanu ◽  
M.C.M. van Loosdrecht
Keyword(s):  
Structure Formation ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Experimental Conditions ◽  
Microscopy Imaging ◽  
Biofilm Growth ◽  
Biofilm Development ◽  
Biofilm Structure

The mathematical modeling of spatial biofilm formation that provides the capability to predict biofilm structure from first principles has been in development for the past six years. However, a direct and quantitative link between model predictions and the experimentally observed structure formation still remains to be established. This work assesses the capability of a state-of-the-art technique for three-dimensional (3D) modeling of biofilm structure, individual based modeling (IbM), to quantitatively describe the early development of a multispecies denitrifying biofilm. Model evaluation was carried out by comparison of predicted structure with that observed from two experimental datasets using confocal laser scanning microscopy (CLSM) monitoring of biofilm development in laboratory flowcells. Experimental conditions provided biofilm growth without substrate limitation, which was confirmed from substrate profiles computed by the model. 3D structures were compared quantitatively using a set of morphological parameters including the biovolume, filled-space profiles, substratum coverage, average thickness and normalized roughness. In spite of the different morphologies detectable in the two independent short-term experiments analyzed here, the model was capable of accurate fitting data from both experiments. Prediction of structure formation was precise, as expressed by the set of morphology parameters used.

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