Journal articles: 'CLSM (confocal laser scanning microscopy)'

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Raarup, Merete Krog, and Jens Randel Nyengaard. "QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY." Image Analysis & Stereology 25, no. 3 (May 3, 2011): 111. http://dx.doi.org/10.5566/ias.v25.p111-120.

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This paper discusses recent advances in confocal laser scanning microscopy (CLSM) for imaging of 3D structure as well as quantitative characterization of biomolecular interactions and diffusion behaviour by means of one- and two-photon excitation. The use of CLSM for improved stereological length estimation in thick (up to 0.5 mm) tissue is proposed. The techniques of FRET (Fluorescence Resonance Energy Transfer), FLIM (Fluorescence Lifetime Imaging Microscopy), FCS (Fluorescence Correlation Spectroscopy) and FRAP (Fluorescence Recovery After Photobleaching) are introduced and their applicability for quantitative imaging of biomolecular (co-)localization and trafficking in live cells described. The advantage of two-photon versus one-photon excitation in relation to these techniques is discussed.

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Zucker, Robert M. "Confocal Microscopy System Performance: Axial Resolution." Microscopy Today 12, no. 1 (January 2004): 38–40. http://dx.doi.org/10.1017/s1551929500051816.

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The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. When tests are made to evaluate the performance of a CLSM, the usual subjective assessment is accomplished by using a histological test slide to create a “pretty picture.” Without the use of functional tests many of the machines may be working at sub-optimal performance levels, delivering sub optimum performance, and possibly misleading data. In order to replace the subjectivity in evaluating a confocal microscope, tests were derived or perfected that measure field illumination, lens clarity, laser power, laser stability, dkhroic functionality, spectral registration, axial resolution, scanning stability, PMT quality, overall machine stability, and system noise (1-3). It is anticipated by using this type of test data, performance standards for confocal microscopes will be obtained and the current subjectivity in evaluating CLSM performance will be eliminated.

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Zucker, Robert M. "Confocal Microscopy System Performance: Field Illumination." Microscopy Today 10, no. 5 (September 2002): 8–13. http://dx.doi.org/10.1017/s1551929500058284.

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The confocal laser-scanning microscope (CLSM) has enormous potential in many biological fields. The reliability of the CLSM to obtain specific measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. For many applications it is useful to know the CLSM system's performance prior to acquiring data images so the necessary resolution, sensitivity and precision can be obtained. Applications involving deconvolution, FRET and quantification necessitate that the confocal microscope is correctly configured and operating at the highest performance levels.

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Chladil, Ladislav, Hana Hálová, and Ondřej Čech. "In-situ Confocal Laser Microscopy Study of Lead Sulfate Crystal Growth on Negative Electrode of Lead-acid Batteries." ECS Transactions 105, no. 1 (November 30, 2021): 159–66. http://dx.doi.org/10.1149/10501.0159ecst.

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Confocal Laser Scanning Microscopy (CLSM) is a widely used technique mainly in fields of biology or multidisciplinary material sciences. Although CLSM has the ability to monitor also electrochemical processes like lead sulfate-crystal growth, nobody used CLSM for such application. We performed operando observation of the pasted active mass of negative electrode for lead-acid batteries during deep cycling. Electrode with pasted negative active mass was optimized for cycling in ECC-opto-std electrochemical cell by EL-CELL. Lead sulfate crystal growth and changes of electrode surface during cycling were observed using a laser scanning confocal microscope Olympus Lext OLS4100. We evaluate the surface changes and sulfate crystal growth. The cycling mode leads to fast gradual degradation of the negative electrode and massive growth of lead sulfate crystals. Confocal laser scanning microscopy was identified as a powerful technique for visualization of lead sulfate crystal promotion during battery cycling.

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Balzano, Angela, Klemen Novak, Miha Humar, and Katarina Čufar. "Application of confocal laser scanning microscopy in dendrochronology." Les/Wood 68, no. 2 (December 30, 2019): 5–17. http://dx.doi.org/10.26614/les-wood.2019.v68n02a01.

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We used the Confocal Laser Scanning Microscope (CLSM) Olympus LEXT OLS5000 for non-destructive observation and image analysis of wood anatomy traits in growth layers of tree species from different climatic zones. In European beech (Fagus sylvatica), where tree rings can generally be recognised, we discuss the changes in tree-ring structure due to adverse effects (insect attacks). Growth layers in Mediterranean Aleppo pine (Pinus halepensis) from south-eastern Spain are not always annual and contain numerous intra-annual density fluctuations (IADFs). Ocote pine (Pinus oocarpa) growing at high elevation in Honduras showed growth layers with clear growth ring boundaries and IADFs. In both pines, CLSM allowed us to recognise and measure tracheid parameters to define density fluctuations. In tropical true mahogany (Swietenia macrophylla) from Venezuela and cedrela (Cedrela odorata) from Costa Rica, we studied the growth layers with variable dimensions of vessels demarcated by marginal axial parenchyma.

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Ockleford, Colin. "The confocal laser scanning microscope (CLSM)." Journal of Pathology 176, no. 1 (May 1995): 1–2. http://dx.doi.org/10.1002/path.1711760102.

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Čufar, Katarina, Angela Balzano, Luka Krže, and Maks Merela. "Wood identification using non-destructive confocal laser scanning microscopy." Les/Wood 68, no. 2 (December 30, 2019): 19–29. http://dx.doi.org/10.26614/les-wood.2019.v68n02a02.

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Exact wood identification is usually based on observation of wood features under the microscope. For this, we have to take a sample of the wood from the object and cut thin slides, possibly of all three anatomical sections. Such destructive sampling is often not possible on valuable historical objects, and therefore there is a need for non-destructive approaches. The objective of the study is to present the potential of Confocal Laser Scanning Microscopy (CLSM) using an Olympus LEXT OLS5000 for the identification of wood. We present work on an example of a gothic sculpture, “St. George Defeating the Dragon”. Conventional sampling and microscopical wood identification showed that St. George is made of Norway spruce (Picea abies), and the dragon of poplar (Populus sp.) or willow (Salix sp.). We present crucial features needed for the identification of these species and the limitations with identification if the samples are too small. Finally, we demonstrate the possibility of wood identification of the abovementioned species using CLSM on wood samples without special preparation of the surfaces. CLSM enabled us to observe all the features needed for wood identification.

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Kubanova, A. A., V. V. Chikin, YU YU Shtirshneider, and O. R. Katunina. "Confocal laser scanning microscopy in vivo for diagnosing melanocytic skin neoplasms." Vestnik dermatologii i venerologii 90, no. 3 (June 24, 2014): 85–94. http://dx.doi.org/10.25208/0042-4609-2014-90-3-85-94.

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The authors discuss the use of confocal laser scanning microscopy in vivo (CLSM) for diagnosing melanocytic skin neoplasms and its value for early diagnostics of melanoma. CLSM is an innovation noninvasive visual examination method for real-time multiple and painless examinations of the patient’s skin without injuring the skin integument. The method ensures early diagnostics of skin melanomas with high sensitivity and specificity, which makes it possible to use CLSM for screening melanocytic skin neoplasms for the sake of the early onset of treatment to save patient life and health.

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Pioch, T., S. Stotz, H. J. Staehle, and H. Duschner. "Applications of Confocal Laser Scanning Microscopy to Dental Bonding." Advances in Dental Research 11, no. 4 (November 1997): 453–61. http://dx.doi.org/10.1177/08959374970110041201.

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The introduction of confocal laser scanning microscopy (CLSM) has provided a valuable new technique for the visualization of bonding structures such as a hybrid layer in dentin (Watson, 1989, 1991), In the case of seven commercially-available dentin bonding systems, it could be demonstrated that the CLSM renders considerably more detailed information than the SEM because of its nondestructive nature and because of the possibility of a distinction between components of bonding agents. With most of the bonding systems, measurements of the thickness of the hybrid layer could be carried out when the primer component was labeled with rhodamine B. It was found that this thickness is significantly increased by increases in etching time and only slightly decreased by increases in the drying time of the dentin and of the primer. When rhodamine B was used for dye penetration tests on four different dentin bonding systems, a leakage within the demineralized zone in the dentin was found in each of the specimens. This structure appears similar to that which Sano et al. (1995) called "nanoleakage". The amount of nanoleakage could not be measured by this method. In the case of enamel or ceramic bonding, a penetration zone was found which corresponded to the etching patterns found in enamel and ceramics, respectively. We conclude that CLSM can offer a wealth of new information about bonding morphology and, therefore, should be used in addition to conventional methods so that the maximum information can be obtained.

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Kim, Yoon Soo, and Adya Singh. "Imaging Degraded Wood by Confocal Microscopy." Microscopy Today 6, no. 4 (May 1998): 14–15. http://dx.doi.org/10.1017/s1551929500067225.

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The application of confocal laser scanning microscopy (CLSM) in the studies of biological materials is rapidly expanding because of the opportunity to produce sharp, high resolution images through optical sectioning and computer assisted 3-D reconstruction. At our institute CLSM is being used in a wide range of forestry and wood science studies.Recently we investigated the potential usefulness of CLSM in characterizing biologically degraded wood. The following are images produced from an archaeological wood which has been buried in a wet environment (rice field) for nearly 2,000 years in South Korea and is apparently degraded by bacteria. In an attempt to develop suitable techniques which can be used for routine examination of fragile degraded wood with CLSM, we have compared two different embedding methods for their suitability in preserving the integrity of cells. The embedding media are paraffin wax and LR White resin.

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ITOH, Johbu. "Three-Dimensional Imaging by Confocal Laser Scanning Microscopy(CLSM)." Journal of Advanced Science 13, no. 1/2 (2001): 7–10. http://dx.doi.org/10.2978/jsas.13.7.

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Cui, Jian Jun. "Lateral Resolution Test for Confocal Laser Scanning Microscope." Key Engineering Materials 609-610 (April 2014): 1159–64. http://dx.doi.org/10.4028/www.scientific.net/kem.609-610.1159.

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To meet the performance test for the confocal laser scanning microscope (CLSM) accurately in some specific application occasions, the spatial resolution imaging principle of confocal laser scanning microscope was analyzed theoretically. And the micron line spacing as the measurement standard has tried to investigate the lateral resolution of CLSM in experiment. The value range of the lateral resolution was calculated by the fluctuation state of the output light intensity signal when there is the lateral movement between the objective with sample. At the same time, some reasons for spatial resolution are also been evaluated in theory. Experiments demonstrate that if the value of the line spacing standard is closer to the spatial resolution of CLSM, the standard can be utilized to test the spatial resolution. So we can use a a series of lines spacing standards with different lines spacing values to test the serial effective resolution. And in our experiment, we only measured line spacing standard with 8μm line width and 100μm line pitch with many times by CLSM, and the spatial resolution of the CLSM is obtained about more minimal than 0.3μm by the scanning curves.

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Marin, Marija, N. Jasnic, D. Lakusic, Sonja Duletic-Lausevic, and Lia Ascensao. "The micromorphological, histochemical and confocal analysis of satureja subspicata Bartl. ex vis. glandular trichomes." Archives of Biological Sciences 62, no. 4 (2010): 1143–49. http://dx.doi.org/10.2298/abs1004143m.

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Micromorphology, histochemical and confocal analyses of the trichomes of Satureja subspicata (Bartl. ex Vis.) were carried out using light microscopy, confocal laser scanning electron microscopy (CLSM), and scanning electron microscopy. Non-glandular unbranched and two types of glandular trichomes - peltate and capitate - are described. The results of histochemical tests showed a positive reaction to phenolics, tannins, lipids, acid lipids, pectins and polysaccharides in both types of glandular trichomes. A strong red autofluorescence of the lipophilic and hydrophilic secreted material in glandular trichomes was observed with CLSM.

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Twamley, Shailey Gale, Anke Stach, Heike Heilmann, Berit Söhl-Kielczynski, Verena Stangl, Antje Ludwig, and Agnieszka Münster-Wandowski. "Immuno-Electron and Confocal Laser Scanning Microscopy of the Glycocalyx." Biology 10, no. 5 (May 4, 2021): 402. http://dx.doi.org/10.3390/biology10050402.

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The glycocalyx (GCX), a pericellular carbohydrate rich hydrogel, forms a selective barrier that shields the cellular membrane, provides mechanical support, and regulates the transport and diffusion of molecules. The GCX is a fragile structure, making it difficult to study by transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM). Sample preparation by conventional chemical fixation destroys the GCX, giving a false impression of its organization. An additional challenge is to process the GCX in a way that preserves its morphology and enhanced antigenicity to study its cell-specific composition. The aim of this study was to provide a protocol to preserve both antigen accessibility and the unique morphology of the GCX. We established a combined high pressure freezing (HPF), osmium-free freeze substitution (FS), rehydration, and pre-embedding immunogold labeling method for TEM. Our results showed specific immunogold labeling of GCX components expressed in human monocytic THP-1 cells, hyaluronic acid receptor (CD44) and chondroitin sulfate (CS), and maintained a well-preserved GCX morphology. We adapted the protocol for antigen localization by CLSM and confirmed the specific distribution pattern of GCX components. The presented combination of HPF, FS, rehydration, and immunolabeling for both TEM and CLSM offers the possibility for analyzing the morphology and composition of the unique GCX structure.

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Balzano, Angela, Katarina Čufar, Luka Krže, and Maks Merela. "Wood identification of charcoal with Confocal Laser Scanning Microscopy." Les/Wood 69, no. 2 (December 30, 2020): 21–35. http://dx.doi.org/10.26614/les-wood.2020.v69n02a02.

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Wood identification of barbecue charcoal from commercial packages of three retailers (B1, B2, B3) in Slovenia and Croatia was performed with help of Confocal Laser Scanning Microscopy (CLSM). CLSM enabled us to image key identification features of charcoal wood that were compared with light micrographs of wood from the reference collection. Product B1 contained charcoal made exclusively of beech wood (Fagus sylvatica) and the declaration indicated the address of the producer, in Serbia which allowed traceability of the wood. The selection of wood species in product B2, consisted of red oak (Quercus cerris or Q. rubra), black locust (Robinia pseudoacacia), and cherry (Prunus avium), which could originate from Serbia, and it did not contain tropical wood as stated on the package. Product B3 contained wood from at least four (sub)topical species which could not be exactly identified to species/genus level. The declaration on the product did not allow traceability of wood. As the risks of illegal logging are high for wood of (sub)tropical origin, our results support the initiative that the monitoring of the charcoal trade should be covered by the EUTR - European Timber Regulations.

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Sakagami, Hiroki, Kosuke Tsuda, Junji Matsumura, and Kazuyuki Oda. "Microcracks Occurring During Drying Visualized by Confocal Laser Scanning Microscopy." IAWA Journal 30, no. 2 (2009): 179–87. http://dx.doi.org/10.1163/22941932-90000213.

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The microcracks occurring during drying of wood were visualized under confocal laser scanning microscopy (CLSM). Precise control of relative humidity and temperature in a specialized environment chamber made it possible to acquire sequential images of the wood of Cryptomeria japonica during drying from the water-saturated condition. The images indicated that the microcracks occurred between tracheid and ray parenchyma in the latewood region and the crack tip advanced in both the bark and pith directions. Subsequently, the crack tip expanding towards the bark stopped at the earlywood region through the growth ring boundary. The other tip toward the pith stopped at the earlywood region before reaching the growth ring boundary. Our technique made it possible to generate microcracks and discuss the relationship between moisture content and microcrack formation during drying. We found the CLSM technique to be an effective method for visualizing microcrack propagation with time.

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Wang, Yu-jing, and Min Ke. "Meibomian Glands or Not? Identification of In Vivo and Ex Vivo Confocal Microscopy Features and Histological Correlates in the Eyelid Margin." Journal of Ophthalmology 2020 (June 30, 2020): 1–8. http://dx.doi.org/10.1155/2020/7516286.

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Purpose. In vivo confocal laser scanning microscopy (CLSM) is an emerging diagnostic tool allowing fast and easy microscopic tissue examination. For the diagnostics of pathological eyelid margin lesions, the knowledge of the normal eyelid margin is essential. Methods. We examined 18 eyelid margins of healthy humans using the in vivo CLSM device and 10 samples of healthy eyelid margins from donor sites with ex vivo CLSM and compared the findings to the corresponding histological sections of donor sites. Cross-section images of different depths and depths of different skin appendages were measured. Results. The depth observed by in vivo CLSM is less than 150 μm into the eyelid. Images of the epidermis and superficial dermis skin, appendages including hair follicle, and sebaceous catheters can be captured associated with histopathology and ex vivo confocal microscopy. In correlation with histopathology, we identified different layers of the eyelid margin, different layers of the epidermis, and skin appendages by ex vivo confocal microscopy. Conclusions. The study offers an overview of the in vivo confocal microscopy human eyelid margin characteristics in comparison to the standard histological examination and confirms that in vivo CLSM could not observe the meibomian gland acini structure.

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Krejsová, Jitka, Magdaléna Doleželová, and Alena Vimmrová. "Evaluation of Fine Aggregate Surface and Fracture Surface by Confocal Microscope." Key Engineering Materials 760 (January 2018): 245–50. http://dx.doi.org/10.4028/www.scientific.net/kem.760.245.

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The gypsum mortars with different types of aggregate were studied. The surface roughness of fine aggregates and the fracture surface roughness were evaluated by a confocal laser scanning microscopy. Four gypsum mortars and one gypsum paste were tested. The results from the confocal laser scanning microscopy (CLSM) are compared with the pictures of grain surface taken by a scanning electron microscopy (SEM) and both methods seem to be appropriate for surface evaluation. The influence of the surface aggregate roughness on some gypsum mortar properties is demonstrated.

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Morrow, Johnica Jo, and Christian Elowsky. "Application of Autofluorescence for Confocal Microscopy to Aid in Archaeoparasitological Analyses." Korean Journal of Parasitology 57, no. 6 (December 31, 2019): 581–85. http://dx.doi.org/10.3347/kjp.2019.57.6.581.

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Confocal laser scanning microscopy (CLSM) was used to examine archaeoparasitological specimens from coprolites associated with La Cueva de los Muertos Chiquitos (CMC) located near present-day Durango, Mexico. The eggs for 4 different types of parasites recovered from CMC coprolites were imaged using CLSM to assist with identification efforts. While some of the parasite eggs recovered from CMC coprolites were readily identified using standard light microscopy (LM), CLSM provided useful data for more challenging identifications by highlighting subtle morphological features and enhancing visualization of parasite egg anatomy. While other advanced microscopy techniques, such as scanning electron microscopy (SEM), may also detect cryptic identifying characters, CLSM is less destructive to the specimens. Utilizing CLSM allows for subsequent examinations, such as molecular analyses, that cannot be performed following SEM sample preparation and imaging. Furthermore, CLSM detects intrinsic autofluorescence molecules, making improved identification independent of resource and time-intensive protocols. These aspects of CLSM make it an excellent method for assisting in taxonomic identification and for acquiring more detailed images of archaeoparasitological specimens.

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Kubanova, A. A., A. A. Kubanov, V. A. Smolyannikova, N. V. Gribanov, and YU B. Makhakova. "Diagnostic value of the confocal laser scanning microscopy in vivo." Vestnik dermatologii i venerologii 91, no. 3 (June 24, 2015): 67–74. http://dx.doi.org/10.25208/0042-4609-2015-91-3-67-74.

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The confocal laser scanning microscopy in vivo is a promising study method to visualize cell structures of epidermis and papillary dermis without affecting the skin integrity, which provides for a resolution and contrast similar to those characteristic of the classical histology examination. Goal. To assess the confocal laser scanning microscopy in vivo (CLSM) technique for diagnosing actinic keratosis, psoriasis vulgaris and rosacea vs. the classical histology examination. Study materials. The article describes the results obtained by using the confocal laser scanning microscopy in vivo technique vs. the histology examination in 10 patients with erythematous actinic keratosis, 10 patients with extensive psoriasis and 10 patients with erythematous and papulous rosacea. Results. The article describes diagnostically significant signs of the diseases detected by using the confocal laser scanning microscopy in vivo as well as the potential of this method in terms of diagnosing inflammatory skin diseases.

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Park, Jae Sung, Chang Kyoung Choi, and Kenneth D. Kihm. "Optically sliced micro-PIV using confocal laser scanning microscopy (CLSM)." Experiments in Fluids 37, no. 1 (March 19, 2004): 105–19. http://dx.doi.org/10.1007/s00348-004-0790-6.

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Dürrenberger, Markus B., Stephan Handschin, Béatrice Conde-Petit, and Felix Escher. "Visualization of Food Structure by Confocal Laser Scanning Microscopy (CLSM)." LWT - Food Science and Technology 34, no. 1 (February 2001): 11–17. http://dx.doi.org/10.1006/fstl.2000.0739.

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Grigoryeva, Natalia Yu, and Dina D. Snarskaya. "Fluorescence methods for investigation of cyanobacterial communities during environmental monitoring of water bodies." Issues of modern algology (Вопросы современной альгологии), no. 2(23) (2020): 8–16. http://dx.doi.org/10.33624/2311-0147-2020-2(23)-8-16.

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The applicability of confocal laser scanning microscopy (CLSM) for environmental monitoring of water bodies is demonstrated on several examples. Such CLSM methods as spectral imaging and microscopic spectroscopy of living cyanobacterial cells are considered. It is shown that fluorescence spectroscopy application can facilitate time-consuming process of taxonomic analysis of field samples and to make monitoring of water bodies during cyanobacterial blooms, on-line.

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Sakagami, Hiroki, Junji Matsumura, and Kazuyuki Oda. "Shrinkage of Tracheid Cells With Desorption Visualized by Confocal Laser Scanning Microscopy." IAWA Journal 28, no. 1 (2007): 29–37. http://dx.doi.org/10.1163/22941932-90001615.

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Confocal laser scanning microscopy (CLSM) was applied as a new method of visualizing the shrinkage of wood and its anisotropy. Control of relative humidity and temperature in a specialized environment chamber made it possible to acquire transverse images of tracheids of Akamatsu (Pinus densiflora) from the saturated condition to the dried condition. The shrinkage of tracheid cells was also determined by measuring the tangential diameter of tracheid and lumen, the radial diameter of tracheid and lumen, and the thickness of tangential and radial walls. Moreover, this technique makes it possible to discuss the relationship between moisture content and tracheid cell shape. We found the CLSM technique to be an effective method for visualizing shrinkage of tracheid cells with desorption.

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Jovin, Thomas M., Michel Robert-Nicoud, Donna J. Arndt-Jovin, and Thorsten Schormann. "3-D imaging of cells using a confocal laser scanning microscope and digital image processing." Proceedings, annual meeting, Electron Microscopy Society of America 46 (1988): 96–97. http://dx.doi.org/10.1017/s0424820100102560.

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Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.

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Pavaloiu, Ramona-Daniela, Fawzia Sha’At, Mousa Sha’At, and Gheorghe Nechifor. "Intracellular Uptake Study of Polymeric Nanoparticles Loaded with Cardiovascular Drugs Using Confocal Laser Scanning Microscopy." Chemistry Proceedings 3, no. 1 (November 14, 2020): 140. http://dx.doi.org/10.3390/ecsoc-24-08427.

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Confocal laser scanning microscopy (CSLM) is a powerful microscopic tool that gives valuable morphological and functional information within cells and tissues. CLSM is non-invasive, with high-contrast scanning, a simple and fast sample preparation procedure as well as easy operation. This paper aimed to study the intracellular uptake of polymeric nanoparticles loaded with cardiovascular drugs using confocal laser scanning microscopy. Polymeric nanoparticles were prepared via nanoprecipitation method using poly(lactide-co-glycolide) (PLGA) as a biodegradable polymeric matrix and Pluronic F127 as a stabilizer. A mixture of two cardiovascular drugs—valsartan (an angiotensin II receptor antagonist drug) and amlodipine besylate (a calcium channel blocker)—was loaded in polymeric nanoparticles. The prepared polymeric nanoparticles had sizes lower than 300 nm and narrow dispersity. The cellular uptake of polymeric nanoparticles was investigated by incubating adherent mouse embryo fibroblasts (NIH 3T3) with a suspension of nanoparticles (stained previously with phthalocyanine) at three different time points. Targeted cell compartments were labeled with two fluorophores: Rhodamine B (membrane stain) and Hoechst (nucleic acid stain). Live cell imaging was performed using a confocal microscope Zeiss LSM710 with Zeiss PALM microdissection system. The intracellular uptake of polymeric nanoparticles was revealed by confocal laser scanning microscopy for each incubation time. The results suggest a possible mechanism of endocytosis and clearly a vesicular-based accumulation of the nanoparticles in the cytoplasmatic compartments.

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Johnston, Robert J., and Thomas G. Mason. "Asphaltene Aggregation Studies of Crude Oil Using 3-D Confocal Microscopy." Microscopy and Microanalysis 6, S2 (August 2000): 22–23. http://dx.doi.org/10.1017/s1431927600032608.

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Confocal laser scanning microscopy (CLSM) has been used to generate three dimensional projection maps of less fluorescent domains caused by asphaltene aggregates in the autofluorescent matrix of crude oil. Heavy crude oils contain asphaltene particles resulting in the production of optically observable micron sized asphaltene aggregates. This technique has been employed to determine the volume fraction of aggregated asphaltenes, øagg, and the time evolution of this phenomenon. The measurements cover a range of various concentrations of asphaltene volume fractions of the heavy asphaltenic oil, øm, from øm = 0 to øm = 0.6.Examining crude oils for asphaltenes aggregates using CLSM presents a challenge due to the strong absorption of the oils. This is because some molecules of the crude oil fluoresce when excited by optical laser light used as a source in the CLSM. Other molecules of the crude oils absorb light at similar wavelengths.

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Marin, Marija, Snezana Budimir, Dusica Janosevic, P. D. Marin, Sonja Duletic-Lausevic, and Milica Ljaljevic-Grbic. "Morphology, distribution, and histochemistry of trichomes of Thymus lykae Degen & Jav. (Lamiaceae)." Archives of Biological Sciences 60, no. 4 (2008): 667–72. http://dx.doi.org/10.2298/abs0804667m.

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Micromorphology, distribution, and histochemistry of the trichomes of Thymus lykae were studied using scanning electron microscopy (SEM)and confocal laser scanning electron microscopy (CLSM). The leaves, stem, and calyx bear numerous glandular and non-glandular trichomes. Two types of glandular trichomes are identified - peltate and capitate. Results of histochemical tests showed positive reactions to polysaccharides, proteins, and lipids. Yellow autofluorescence of secreted material was noticed in peltate and capitate trichomes. Strong reddish-yellow autofluorescence of the lipophilic and hydrophilic secreted material was observed with CLSM.

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Castejόn, O., and P. Sims. "Light Microscopy, Confocal Laser Scanning Microscopy And Scanning And Transmission Electron Microscopy Of Cerebellar Golgi Cells." Microscopy and Microanalysis 5, S2 (August 1999): 1302–3. http://dx.doi.org/10.1017/s1431927600019838.

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The cerebellar cortex of albino mice, hamsters, teleost fishes, primates and human have been examined by correlative microscopy to study the Golgi cell soma, dendritic processes, axonal plexus and synaptic connections in the granular and molecular layers. For light microscopy (LM) toluidinc blue stained-plastic embedded scmithin sections and Golgi light microscopy preparations were used. For confocal laser scanning microscopy (CLSM) of hamster cerebellum the FM4-64 fluorescent stain was used as intracellular tracer (1). Conventional and high resolution scanning electron microscopy (SEM) of teleost fishes, primates and human were coated with gold-palladium and chromium (2). I Transmission electron microscopy (TEM). either by ullrathin sections or frccze-clching replicas, were examined to characterize synaptic connections in the granular and molecular layers. The Golgi cells appeared in the granular layer as polygonal, stellate, round or fusiform microncurons. 10-25 μm in maximal dimension, surrounded by the granule cell groups. Golgi light microscopy.

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Depypere, F., P. Van Oostveldt, J. G. Pieters, and K. Dewettinck. "Quantification of microparticle coating quality by confocal laser scanning microscopy (CLSM)." European Journal of Pharmaceutics and Biopharmaceutics 73, no. 1 (September 2009): 179–86. http://dx.doi.org/10.1016/j.ejpb.2009.04.007.

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Zucker, Robert M. "Confocal Microscopy System Performance: Laser Power Measurements." Microscopy Today 10, no. 6 (November 2002): 20–23. http://dx.doi.org/10.1017/s1551929500058466.

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The reliability of the confocai laser-scanning microscope (CLSM) to obtain intensity measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. The laser power test appears to be one of the most useful tests to quickly evaluate if a system is misaligned or functioning sub optimally by recording insufficient laser power readings. The test using a power meter can indicate if the system is aligned properly up to the plane of excitation on the stage, or if the machine has a defective component (i.e. a dying laser, or a defective fiber). In our experience, without sufficient power throughput in the system, the PMT voltages will have to be increased to high values to visualize fluorescence derived from specimens, which will introduce reduced image quality. In addition the cause of the decreased laser power may result in other problems i.e. laser instability, loss of axial resolution, and increased noise.

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Schubert, Mark, Chris Stührk, Matthias J. Fuhr, and Francis W. M. R. Schwarze. "Imaging hyphal growth of Physisporinus vitreus in Norway spruce wood by means of confocal laser scanning microscopy (CLSM)." Holzforschung 68, no. 6 (August 1, 2014): 727–30. http://dx.doi.org/10.1515/hf-2013-0183.

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Abstract Light microscopy and electron microscopy are the most common methods for analyzing wood-decay fungi. However, the 3D visualization and quantification of the filamentous structure of fungi in wood is difficult to realize by means of these traditional techniques. In the present work, confocal laser scanning microscopy (CLSM) was further developed for the quantitative imaging of the 3D microscopic hyphal growth of Physisporinus vitreus, a versatile fungus for engineering value-added wood products. To this purpose, the fungus was stained with a fluorescent dye Alexa Fluor. The 3D information obtained by CLSM has a high potential as a basis for the development of mathematical models for a more precise observation of the growth behavior of wood-decay fungi.

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Zucker, Robert M., Susan C. Jeffery, and Sally D. Perreault. "Mammalian Apoptosis in Whole Neonatal Ovaries using Confocal Laser Scanning Microscopy." Microscopy and Microanalysis 7, S2 (August 2001): 16–17. http://dx.doi.org/10.1017/s1431927600026155.

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Confocal Laser Scanning Microscopy (CLSM) is being used to visualize folliculogenesis and presumed apoptotic events in whole neonatal mouse/rat ovaries. Our laboratory has previously demonstrated that apoptosis can be visualized in 8-9 day whole mouse embryos, 12 day fetal mouse eyes or 12-15 fetal rat limbs stained with the fluorescent lysosomal stain, LysoTracker Red (LT). The observed LT staining has been correlated with Nile blue staining (lysosomes and phagocytosis), TUNEL staining (DNA breaks) and apoptotic tissue morphology in rat and mouse developmental systems. This increased LT staining intensity appeared to be associated with acidic tissue, lysosomes, phagocytosis and dying cells. Here we describe recent progress using LT to evaluate the apoptotic process in whole ovaries. We are currently optimizing sample preparation and confocal machine operation to improve the image quality. to increase depth of visualization, ovaries derived from 10-45 day old mice/rats are fixed with an aldehyde and then made transparent by MEOH dehydration followed by benzyl alcohol/benzyl aldehyde (BABB) clearing. Such whole ovaries or tissue sections are viewed by CLSM using 568-wavelength laser light to obtain images of oocytes and follicles. Follicles showing gross morphologic changes typical of atresia were also characterized by intense LT staining of granulosa cells. This increased LT staining intensity appeared to be associated with acidic tissue and dying cells contained within the follicle.

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Szarowski, Donald H., Michael Fejtl, Paul McCauley, David O. Carpenter, and James N. Turner. "Neuron model system: Correlative confocal laser scanned microscopy and membrane physiology." Proceedings, annual meeting, Electron Microscopy Society of America 48, no. 3 (August 12, 1990): 170–71. http://dx.doi.org/10.1017/s042482010015839x.

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Confocal laser scanning microscopy (CLSM) has been used to correlate morphology and membrane physiology in cultured neurons, providing a model system for studying physiologic and pathologic conditions. Ion channels are studied by patch-clamp methods as a function of receptor stimulation and toxic excitatory amino acids, including those implicated in Alzheimer’s dementia. Glial cells are often closely associated with the neurons, and are difficult to detect in living cultures due to the relative sizes of glia and neurons (5-20 μm versus 125 μm), compounded with the fact that they are thick phase objects. Groups of glia can also be confused with neurons. Thus it is difficult to select appropriate cells and/or cell regions for patch-clamping. We are correlating physiology and conventional light microscopy with CLSM to determine the role of glia, and neuron surface geometry on the ability to establish Gigaohm membrane-micropipette seals. Morphology of the system as observed by CLSM is presented here.

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Guida, Stefania, Federica Arginelli, Francesca Farnetani, Silvana Ciardo, Laura Bertoni, Marco Manfredini, Nicola Zerbinati, Caterina Longo, and Giovanni Pellacani. "Clinical Applications of In Vivo and Ex Vivo Confocal Microscopy." Applied Sciences 11, no. 5 (February 24, 2021): 1979. http://dx.doi.org/10.3390/app11051979.

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Confocal laser scanning microscopy (CLSM) has been introduced in clinical settings as a tool enabling a quasi-histologic view of a given tissue, without performing a biopsy. It has been applied to many fields of medicine mainly to the skin and to the analysis of skin cancers for both in vivo and ex vivo CLSM. In vivo CLSM involves reflectance mode, which is based on refractive index of cell structures serving as endogenous chromophores, reaching a depth of exploration of 200 μm. It has been proven to increase the diagnostic accuracy of skin cancers, both melanoma and non-melanoma. While histopathologic examination is the gold standard for diagnosis, in vivo CLSM alone and in addition to dermoscopy, contributes to the reduction of the number of excised lesions to exclude a melanoma, and to improve margin recognition in lentigo maligna, enabling tissue sparing for excisions. Ex vivo CLSM can be performed in reflectance and fluorescent mode. Fluorescence confocal microscopy is applied for “real-time” pathological examination of freshly excised specimens for diagnostic purposes and for the evaluation of margin clearance after excision in Mohs surgery. Further prospective interventional studies using CLSM might contribute to increase the knowledge about its application, reproducing real-life settings.

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CHETVERIKOV, PHILIPP E. "Confocal laser scanning microscopy technique for the study of internal genitalia and external morphology of eriophyoid mites (Acari: Eriophyoidea)." Zootaxa 3453, no. 1 (September 5, 2012): 56. http://dx.doi.org/10.11646/zootaxa.3453.1.4.

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Confocal laser scanning microscopy (CLSM) is a modern powerful technique that can be used for studying the externaland internal anatomy of arthropods. CLSM has seldom been used in acarology and very rarely for studying eriophyoidmites. It allows the capture of precise digital images of the fine details of external and internal chitinous structures, whichcan be further analysed using various computer programs. CLSM can serve as an effective tool for comparing closelyrelated and/or cryptic species, correcting diagnoses of poorly described taxa, studying immature instars, and particularly,for studying the structures and the functioning of the internal genitalia of adult females and males. In this paper, thepotential use of CLSM for the study of eriophyoids is demonstrated using specimens of 13 mite species and eight generafrom the families Phytoptidae Murray 1877 and Eriophyidae Nalepa 1898. This study showed that freshly mountedspecimens on microscope slides appeared to be the most appropriate for CLSM as older specimens tended to have reducedautofluorescence. The best choice for studying the external morphology and internal genital apparatus of eriophyoid mitesappeared to be the blue laser. Green and light blue wavelengths (488 nm and 532 nm) were found to be less useful. Thequality of CLSM images depended on the slide-mounting medium used. Among those compared, Hoyer’s medium wasfound to be the most appropriate whereas Heinze medium and media including Iodium gave poorer results. The empodiaand proximal parts of setae were shown to have very weak autofluorescence signals, but they reflected red (635 nm) and blue (405 nm) laser light, which could be detected with CLSM.

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NIX, THOMAS, and SUSANNE FEIST-BURKHARDT. "New methods applied to the microstructure analysis of Messel oil shale: confocal laser scanning microscopy (CLSM) and environmental scanning electron microscopy (ESEM)." Geological Magazine 140, no. 4 (July 2003): 469–78. http://dx.doi.org/10.1017/s0016756803008094.

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Investigations of mass movements of the Messel oil shale in Germany presumed that swell- and especially shrink-deformations of the organic-rich clay were among the factors triggering initial displacements. Within the scope of the present studies, delicate clay/organic microstructures had to be deciphered. Reactions of clay minerals and microstructures upon dehydration were observed by environmental scanning electron microscopy (ESEM) and the distribution of organic matter was studied by confocal laser scanning microscopy (CLSM) in fluorescence mode. ESEM and CLSM have been demonstrated to be valuable tools for the microstructural characterization of laminated organic-rich sediments. The application of these analytical and imaging techniques will be of great interest to a wide range of geological disciplines such as palaeontology, organic petrology, engineering geology and global change studies.

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Lawrence, J. R., G. D. W. Swerhone, G. G. Leppard, T. Araki, X. Zhang, M. M. West, and A. P. Hitchcock. "Scanning Transmission X-Ray, Laser Scanning, and Transmission Electron Microscopy Mapping of the Exopolymeric Matrix of Microbial Biofilms." Applied and Environmental Microbiology 69, no. 9 (September 2003): 5543–54. http://dx.doi.org/10.1128/aem.69.9.5543-5554.2003.

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ABSTRACT Confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and soft X-ray scanning transmission X-ray microscopy (STXM) were used to map the distribution of macromolecular subcomponents (e.g., polysaccharides, proteins, lipids, and nucleic acids) of biofilm cells and matrix. The biofilms were developed from river water supplemented with methanol, and although they comprised a complex microbial community, the biofilms were dominated by heterotrophic bacteria. TEM provided the highest-resolution structural imaging, CLSM provided detailed compositional information when used in conjunction with molecular probes, and STXM provided compositional mapping of macromolecule distributions without the addition of probes. By examining exactly the same region of a sample with combinations of these techniques (STXM with CLSM and STXM with TEM), we demonstrate that this combination of multimicroscopy analysis can be used to create a detailed correlative map of biofilm structure and composition. We are using these correlative techniques to improve our understanding of the biochemical basis for biofilm organization and to assist studies intended to investigate and optimize biofilms for environmental remediation applications.

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Abe, Hisashi, Ryo Funada, Naohiro Kuroda, Osamu Furusawa, Masayuki Shibagaki, and Tomoyuki Fujii. "CONFOCAL LASER SCANNING MICROSCOPY OF WATER UPTAKE DURING THE RECOVERY OF COMPRESSED AND DRYING-SET WOOD." IAWA Journal 22, no. 1 (2001): 63–72. http://dx.doi.org/10.1163/22941932-90000269.

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The initial uptake of water by small fragments of compressed and drying- set wood of Cryptomeria japonica D. Don was monitored by confocal laser scanning microscopy (CLSM) using an aqueous solution of the fluorescent dye acridine orange. CLSM allowed visualization of the recovery over time of compressed and drying-set wood and the uptake of water by the specimens. Furthermore, CLSM allowed us to monitor the structure of deformed tracheids under atmospheric conditions. Increases in the compression ratio increased the time required for the uptake of water. The uptake of water was detected first between deformed and undeformed regions of compressed and drying-set wood at all compression ratios tested.

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Haupt, B. J., A. E. Pelling, and M. A. Horton. "Integrated Confocal and Scanning Probe Microscopy for Biomedical Research." Scientific World JOURNAL 6 (2006): 1609–18. http://dx.doi.org/10.1100/tsw.2006.269.

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Atomic force microscopy (AFM) continues to be developed, not only in design, but also in application. The new focus of using AFM is changing from pure material to biomedical studies. More frequently, it is being used in combination with other optical imaging methods, such as confocal laser scanning microscopy (CLSM) and fluorescent imaging, to provide a more comprehensive understanding of biological systems. To date, AFM has been used increasingly as a precise micromanipulator, probing and altering the mechanobiological characteristics of living cells and tissues, in order to examine specific, receptor-ligand interactions, material properties, and cell behavior. In this review, we discuss the development of this new hybrid AFM, current research, and potential applications in diagnosis and the detection of disease.

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Ruini, Cristel, Sophia Schlingmann, Žan Jonke, Pinar Avci, Víctor Padrón-Laso, Florian Neumeier, Istvan Koveshazi, et al. "Machine Learning Based Prediction of Squamous Cell Carcinoma in Ex Vivo Confocal Laser Scanning Microscopy." Cancers 13, no. 21 (November 3, 2021): 5522. http://dx.doi.org/10.3390/cancers13215522.

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Image classification with convolutional neural networks (CNN) offers an unprecedented opportunity to medical imaging. Regulatory agencies in the USA and Europe have already cleared numerous deep learning/machine learning based medical devices and algorithms. While the field of radiology is on the forefront of artificial intelligence (AI) revolution, conventional pathology, which commonly relies on examination of tissue samples on a glass slide, is falling behind in leveraging this technology. On the other hand, ex vivo confocal laser scanning microscopy (ex vivo CLSM), owing to its digital workflow features, has a high potential to benefit from integrating AI tools into the assessment and decision-making process. Aim of this work was to explore a preliminary application of CNN in digitally stained ex vivo CLSM images of cutaneous squamous cell carcinoma (cSCC) for automated detection of tumor tissue. Thirty-four freshly excised tissue samples were prospectively collected and examined immediately after resection. After the histologically confirmed ex vivo CLSM diagnosis, the tumor tissue was annotated for segmentation by experts, in order to train the MobileNet CNN. The model was then trained and evaluated using cross validation. The overall sensitivity and specificity of the deep neural network for detecting cSCC and tumor free areas on ex vivo CLSM slides compared to expert evaluation were 0.76 and 0.91, respectively. The area under the ROC curve was equal to 0.90 and the area under the precision-recall curve was 0.85. The results demonstrate a high potential of deep learning models to detect cSCC regions on digitally stained ex vivo CLSM slides and to distinguish them from tumor-free skin.

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Singh, Adya, Ying Xiao, and Robin Wakeling. "Glutaraldehyde Autofluorescence Useful in Confocal Studies of Fungi." Microscopy Today 5, no. 8 (October 1997): 16–17. http://dx.doi.org/10.1017/s1551929500056753.

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In this note we show the potential usefulness of glutaraldehyde (GA) in confocai microscopic studies of wood-fungal interaction. We are presently developing methods to examine by the confocai laser scanning microscope (CLSM), the pattern of distribution of fungal hyphae within wood in relation to fungal degradation and sapstaining of wood. CLSM has the potential to be a very useful tool in such studies for a variety of reasons, including its use for optical sectioning to produce computer assisted 3-D images.

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Göring, Lena, Markus Finkeldey, Falk Schellenberg, Carsten Brenner, Martin R. Hofmann, and Nils C Gerhardt. "Optical metrology for the investigation of buried technical structures." tm - Technisches Messen 85, no. 2 (February 23, 2018): 104–10. http://dx.doi.org/10.1515/teme-2017-0096.

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Abstract In this paper, we present different optical metrology approaches for the investigation of buried technical structures. Contactless, potentially fast and non-destructive techniques such as optical beam induced current (OBIC), confocal laser scanning microscopy (CLSM) and digital holographic microscopy (DHM) are described. Their properties are illustrated by investigating the buried structures of a microcontroller.

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Nishikawa, Tetsunari, Kazuya Masuno, Masahiko Mori, Yasuhiro Tajime, Kenji Kakudo, and Akio Tanaka. "Calcification at the Interface Between Titanium Implants and Bone: Observation With Confocal Laser Scanning Microscopy." Journal of Oral Implantology 32, no. 5 (October 1, 2006): 211–17. http://dx.doi.org/10.1563/799.1.

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Abstract It has not been previously possible to observe bone formation in undecalcified sections with titanium implants at high magnification because of the difficulty in sectioning bone together with implants. A method for examining the bone-implant interface in undecalcified sections is described in which implants are left in situ and confocal laser scanning microscopy (CLSM) is used to examine both the implant surface and adjacent bone. Pulsing of animals at different times with the fluorescent dyes calcein and alizarin red permitted assessment of temporal patterns of bone formation by CLSM. Reflectivity of the polished implant surface permitted accurate assessment of the position of the implant relative to labeled bone. The analysis showed that bone first formed as thin processes towards and across the implant surface, followed by further bone formation behind these processes. The interface between calcified bone tissue and the implant surface was characterized by a 10-μm space. The CLSM technique enabled detailed observations of new bone formation at the titanium implant interface.

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Mohandas, Laxmi, Anju T. R., and Sarita G. Bhat*. "Confocal and SEM imaging to demonstrate food pathogen- biofilm biocontrol by pyocyanin from Pseudomonas aeruginosa BTRY1." International Journal of Bioassays 6, no. 01 (December 31, 2016): 5218. http://dx.doi.org/10.21746/ijbio.2017.01.007.

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An assortment of redox-active phenazine compounds like pyocyanin with their characteristic blue-green colour are synthesized by Pseudomonas aeruginosa, Gram-negative opportunistic pathogens, which are also considered one of the most commercially valuable microorganisms. In this study, pyocyanin from Pseudomonas aeruginosa BTRY1 from food sample was assessed for its antibiofilm activity by micro titer plate assay against strong biofilm producers belonging to the genera Bacillus, Staphylococcus, Brevibacterium and Micrococcus. Pyocyanin inhibited biofilm activity in very minute concentrations. This was also confirmed by Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM). Both SEM and CLSM helped to visualize the biocontrol of biofilm formation by eight pathogens. The imaging and quantification by CLSM also established the impact of pyocyanin on biofilm-biocontrol mainly in the food industry.

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Neilly, J. P., G. D. Gagne, J. Bryant, B. Daanen, and K. Schuette. "Applications of Confocal Microscopy and Scanning Electron Microscopy to the Localization of Immunoreagents Used in Medical Diagnostic Systems." Proceedings, annual meeting, Electron Microscopy Society of America 54 (August 11, 1996): 306–7. http://dx.doi.org/10.1017/s042482010016399x.

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Medical diagnostic assays often are based on the immobilization of immunolabeling reagents on solid substrates such as polystyrene beads, microparticles, or membranes. The distribution of immunoreagents on or within these substrates has a significant effect on product performance. Confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) can be used to localize immunological reagents on beads and other surfaces. In this paper we describe examples in which CLSM and SEM were used to assist in the design and troubleshooting of three diagnostic systems.A protective overcoat is used on HIV antibody-coated 0.25“ polystyrene beads in an HIV screening assay. To visualize topographical detail of the antibody distribution on beads with and without the overcoat, beads were labeled with a fluorescent secondary antibody and examined by CLSM. On overcoated beads, the distribution of HIV antibody was relatively uniform; but on non-overcoated beads, HIV antibody distribution was patchy and located mostly in low areas of the bead surface (Fig. 1), evidence that the overcoat provided protection for the HIV antibody. In the same assay system, performance was enhanced by using beads coated with a mixture of HIV antibodies from two IgG subclasses. The relative distribution of the two primary antibodies on the bead surface was demonstrated using isotype-specific secondary antibodies and CLSM.

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Mallya, Laxmish, M. Kundabala, and Vinod Jathanna. "Confocal LASER Scanning Microscopy (CLSM) for Evaluation of Endodontic Microflora-A Review." Indian Journal of Public Health Research & Development 10, no. 1 (2019): 69. http://dx.doi.org/10.5958/0976-5506.2019.00015.9.

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Ozaki, Yasushi. "Application of Confocal Laser Scanning Microscopy (CLSM) for Observing Adhesives in Paper." Journal of Adhesion Science and Technology 25, no. 6-7 (January 2011): 723–41. http://dx.doi.org/10.1163/016942410x525902.

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Hanthamrongwit, M., M. H. Grant, and R. Wilkinson. "Confocal laser scanning microscopy (CLSM) for the study of collagen sponge microstructure." Journal of Biomedical Materials Research 28, no. 2 (February 1994): 213–16. http://dx.doi.org/10.1002/jbm.820280211.

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Tzanakakis, Emmanouhl S., Chang-Chun Hsiao, Taku Matsushita, Rory P. Remmel, and Wei-Shou Hu. "Probing Enhanced Cytochrome P450 2B1/2 Activity in Rat Hepatocyte Spheroids through Confocal Laser Scanning Microscopy." Cell Transplantation 10, no. 3 (April 2001): 329–42. http://dx.doi.org/10.3727/000000001783986783.

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Cytochrome P450 (CYP450) enzymes are essential for xenobiotic metabolism. Although CYP450s are found in many tissues, CYP2B1/2 are primarily expressed in the rat liver. The constitutive expression in vivo of CYP2B1/2 is low but it is induced in the presence of various drugs such as phenobarbital (PB). In this study, CYP2B1/2 activity in cultured hepatocytes was assessed in situ with the introduction of a fluorogenic sub-strate, pentoxyresorufin. The product of 7-pentoxyresorufin-O-dealkylation (PROD), which is catalyzed specifically by CYP2B1/2, was detected using confocal laser scanning microscopy (CLSM). Primary hepatocytes cultured as monolayers on collagen-coated surfaces exhibited background PROD activity and minimal PB inducibility after 4 days in culture. In contrast, rat hepatocytes organized in compacted aggregates, or spheroids, exhibited higher levels of PROD activity and retained their ability for PB induction. The results from the CLSM analysis were verified by RT-PCR and Western immunoblotting analysis. Furthermore, CLSM in conjunction with image processing techniques and three-dimensional reconstruction revealed the localization of enhanced PROD activity in the center of spheroids. The results support the use of CLSM as a powerful tool for investigating CYP2B1/2 activity in cultured rat hepatocytes.

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Journal articles on the topic 'CLSM (confocal laser scanning microscopy)'

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