Page Baluch

Arizona State University, School of Life Sciences, Manager WM Keck Bioimaging Facility

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Tel Aviv University

Northwestern University

Liliya Gabelev

Renping Zhou

Rutgers, The State University of New Jersey

Ilpo Huhtaniemi

Xuefeng Xia

Ray Wong

National University of Singapore

Sook-young Yoon

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Papers by Page Baluch

The Role of Tyramine Within the Mouse Ovary

Microscopy and Microanalysis, 2019

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Methods for Creating and Evaluating 3D Tactile Images to Teach Stem Courses to the Visually Impaired

National Science Teachers Association, Jul 1, 2015

effectiveness of incorporating 3D tactile images, which are critical for learning science, techno... more effectiveness of incorporating 3D tactile images, which are critical for learning science, technology, engineering, and mathematics, into entry-level lab courses. The focus of this project is to increase the participation and performance of blind or visually impaired (BVI) students by providing a multimodal tactile approach to learning image-rich material. This study explored the use of various tactile image formats and activity sets to evaluate their ability to assist students in lab exercise. These haptic tools were evaluated in classes that had both sighted and BVI students as well as in participation studies comprised primarily of individuals with vision impairment. The results of this study demonstrated that the inclusion of tactile images is helpful to both sighted and vision-impaired students and should be explored further to determine the extent of their educational benefit.

Volume Segmentation Using Weibull ESD Fields

This paper presents a coarse-grain approach based on the Weibull E-SD field for segmentation of a... more This paper presents a coarse-grain approach based on the Weibull E-SD field for segmentation of an object with gray levels appearing in a volume data. The input data is on a 3D structured grid and of the form δ =) , , (k j i v , where δ is some scalar value. In this paper, we consider a voxel as a k k k × × cube, and each voxel is assigned two values: expectancy and standard deviation (E-SD). We use the Weibull noise index to estimate the noise in a voxel, and to obtain more precise E-SD values for each voxel. Our test bed includes synthetic data as well as real volume data from a scanning laser confocal microscope. Analysis of this data all show distinct and defining regions in their E-SD plot. Under the guide of an E-SD plot, we can efficiently segment an object embedded in real and simulated 3D data.

Printing Tactile Graphics for the Blind : Application to Histology

Visual images, whether graphs, charts, diagrams, photographs, models, or actual specimens, have a... more Visual images, whether graphs, charts, diagrams, photographs, models, or actual specimens, have always been an important part of the educational process as an efficient way of conveying information. This is particularly true for science disciplines. Indeed, in today’s world of rapidly proliferating information, in an effort to concisely present this information, the trend is toward more stand-alone graphics. According to Burch and Pawluk (2011), over 70% of textbooks contain diagrams with no accompanying description in the text. This means that paradoxically while the number of electronically available textbooks is greatly increasing, which should lead to increased access for people who are blind or visually impaired (BVI), the reliance on graphical representations causes the information to be more inaccessible.

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Fibrin polymer on the surface of biomaterial implants drives the foreign body reaction

Biomaterials

Tyramine Localization Closely Corelates to Circular Vesicles Within the Mouse Uterine Horn Using Correlational Fluorescence and Scanning Electron Microscopy

Microscopy and Microanalysis

This is an Accepted Manuscript for the Microscopy and Microanalysis 2020 Proceedings. This versio... more

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An actin-based protrusion originating from a podosome-enriched region initiates macrophage fusion

Molecular Biology of the Cell

Macrophage fusion resulting in the formation of multinucleated giant cells occurs in a variety of... more Macrophage fusion resulting in the formation of multinucleated giant cells occurs in a variety of chronic inflammatory diseases, yet the mechanism responsible for initiating this process is unknown. Here, we used live cell imaging to show that actin-based protrusions at the leading edge initiate macrophage fusion. Phase-contrast video microscopy demonstrated that in the majority of events, short protrusions (∼3 µm) between two closely apposed cells initiated fusion, but occasionally we observed long protrusions (∼12 µm). Using macrophages isolated from LifeAct mice and imaging with lattice light sheet microscopy, we further found that fusion-competent protrusions formed at sites enriched in podosomes. Inducing fusion in mixed populations of GFP- and mRFP-LifeAct macrophages showed rapid spatial overlap between GFP and RFP signal at the site of fusion. Cytochalasin B strongly reduced fusion and when rare fusion events occurred, protrusions were not observed. Fusion of macrophages def...

GSK3� mediates acentromeric spindle stabilization by activated PKC?

Develop Biol, 2008

Jumpstarting STEM Careers

Forward to Professorship in STEM, 2016

<title>Line and net pattern segmentation using shape modeling</title>

Visualization and Data Analysis 2003, 2003

ABSTRACT Line and net patterns in a noisy environment exist in many biomedical images. Examples i... more ABSTRACT Line and net patterns in a noisy environment exist in many biomedical images. Examples include: Blood vessels in angiography, white matter in brain MRI scans, and cell spindle fibers in confocal microscopic data. These piecewise linear patterns with a Gaussian-like profile can be differentiated from others by their distinctive shape characteristics. A shape-based modeling method is developed to enhance and segment line and net patterns. The algorithm is implemented in an enhancement/thresholding type of edge operators. Line and net features are enhanced by second partial derivatives and segmented by thresholding. The method is tested on synthetic, angiography, MRI, and confocal microscopic data. The results are compared to the implementation of matched filters and crest lines. It shows that our new method is robust and suitable for different types of data in a broad range of noise levels.

The Arabidopsis thaliana putative sialyltransferase resides in the Golgi apparatus but lacks the ability to transfer sialic acid

Plant Biology, 2009

A common feature of the animal sialyltransferases (STs) is the presence of four conserved motifs,... more A common feature of the animal sialyltransferases (STs) is the presence of four conserved motifs, namely large (L), small (S), very small (VS) and motif III. Although sialic acid (SA) has not been detected in plants, three orthologues containing sequences similar to the ST motifs have been identified in the Arabidopsis thaliana L. database. In this study, we report that the At3g48820 gene (Gene ID: 824043) codes for a Golgi resident protein lacking the ability to transfer SA to asialofetuin or Galb1,3GalNAc and Galb1,4Glc-NAc oligosaccharide acceptors. Restoration of deteriorated motifs S, VS and motif III by constructing chimeric proteins consisting of the 28-308 amino acid region of the A. thaliana At3g48820 ST-like protein and the 264-393 amino acid region of the Oryza sativa L. AK107493 ST-like protein, or of the 28-240 amino acid region of the At3g48820 protein and the 204-350 amino acid region of the Homo sapiens L. a2,3-ST (NP_008858) was not able to recover sialyltransferase activity. Altering the appropriate amino acid regions of the A. thaliana At3g48820 ST-like protein to those typical for the mammalian motif III (HHYWE) and VS motif (HDADFE) also did not have any effect. Our data, together with previous results, indicate that A. thaliana in particular, and plants in general, do not have transferases for SA. Substrates for the plant ST-like proteins might be compounds involved in secondary metabolism.

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A semi-automated analysis method of small sensory nerve fibers in human skin-biopsies

Journal of Neuroscience Methods, 2010

Computerized detection method (CDM) software programs have been extensively developed in the fiel... more Computerized detection method (CDM) software programs have been extensively developed in the field of astronomy to process and analyze images from nearby bright stars to tiny galaxies at the edge of the Universe. These object-recognition algorithms have potentially broader applications, including the detection and quantification of cutaneous small sensory nerve fibers (SSNFs) found in the dermal and epidermal layers, and in the intervening basement membrane of a skin punch biopsy. Here, we report the use of astronomical software adapted as a semi-automated method to perform density measurements of SSNFs in skin-biopsies imaged by Laser Scanning Confocal Microscopy (LSCM). In the first half of the paper, we present a detailed description of how the CDM is applied to analyze the images of skin punch biopsies. We compare the CDM results to the visual classification results in the second half of the paper. Abbreviations used in the paper, description of each astronomical tools, and their basic settings and how-tos are described in the appendices. Comparison between the normalized CDM and the visual classification results on identical images demonstrates that the two density measurements are comparable. The CDM therefore can be used-at a relatively low cost-as a quick (a few hours for entire processing of a single biopsy with 8-10 scans) and reliable (high-repeatability with minimum user-dependence) method to determine the densities of SSNFs.

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Identification of a Role for CLASP2 in Insulin Action

Journal of Biological Chemistry, 2012

Background: Insulin stimulates glucose uptake in target tissues to promote blood glucose homeosta... more Background: Insulin stimulates glucose uptake in target tissues to promote blood glucose homeostasis, although the mechanism is not fully understood. Results: siRNA-mediated knockdown of CLASP2 reduces insulin-stimulated glucose transport. Conclusion: CLASP2 is a new protein involved in insulin-stimulated glucose uptake. Significance: The identification of CLASP2 as a new player in insulin action provides an improved understanding of insulinstimulated glucose uptake and GLUT4 trafficking.

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Involvement of PKCζ and GSK3β in the stability of the metaphase spindle

In Vitro Cellular & Developmental Biology - Animal, 2012

In the somatic cell, the mitotic spindle apparatus is centrosomal, and several isoforms of protei... more In the somatic cell, the mitotic spindle apparatus is centrosomal, and several isoforms of protein kinase C (PKC) have been associated with the mitotic spindle, but their role in stabilizing the mitotic spindle is still unclear. Other protein kinases such as, glycogen synthase kinase 3β (GSK3β) have also been shown to be associated with the mitotic spindle apparatus. In this study, we show the enrichment of active (phosphorylated) PKCζ at the centrosomal region of the spindle apparatus in metaphase stage of 3T3 cells. In order to understand whether the two kinases PKC and GSK3β are associated with the mitotic spindle, first, the co-localization of phosphorylated PKC isoforms with GSK3β was studied at the poles in metaphase cells. Fluorescence resonance energy transfer (FRET) analysis was used to demonstrate close molecular proximity of phospho-PKCζ with phospho(ser9)GSK3β. Second, the involvement of inactive GSK3β in maintaining an intact mitotic spindle in 3T3 cells was shown. Third, this study also showed that addition of a phospho-PKCζ specific inhibitor to cells can disrupt the mitotic spindle microtubules and some of the proteins associated with it. The mitotic spindle at metaphase in mouse fibroblasts appears to be maintained by PKCζ acting through GSK3β. Phospho-PKCζ is in close molecular proximity to GSK3β, whereas the other isoforms of PKC such as pPKCβII, pPKCγ, pPKCμ, and pPKCθ are not close enough to have significant FRET readings. The close molecular proximity supports the idea that GSK3β may be a substrate of PKCζ.

Thin structure segmentation and visualization in three-dimensional biomedical images: a shape-based approach

IEEE Transactions on Visualization and Computer Graphics, 2000

This paper presents a shape-based approach in extracting thin structures, such as lines and sheet... more This paper presents a shape-based approach in extracting thin structures, such as lines and sheets, from threedimensional (3D) biomedical images. Of particular interest is the capability to recover cellular structures, such as microtubule spindle fibers and plasma membranes, from laser scanning confocal microscopic (LSCM) data. Hessian-based shape methods are reviewed. A synthesized linear structure is used to evaluate the sensitivity of the multiscale filtering approach in extracting closely positioned fibers. We find that the multiscale approach tends to fuse lines together, which makes it unsuitable for visualizing mouse egg spindle fibers. Single-scale Gaussian filters, balanced between sensitivity and noise resistance, are adopted instead. In addition, through an ellipsoidal Gaussian model, the eigenvalues of the Hessian matrix are quantitatively associated with the standard deviations of the Gaussian model. Existing shape filters are simplified and applied to LSCM data. A significant improvement in extracting closely positioned thin lines is demonstrated by the resultant images. Further, the direct association of shape models and eigenvalues makes the processed images more understandable qualitatively and quantitatively.

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On marching cubes

IEEE Transactions on Visualization and Computer Graphics, 2003

A characterization and classification of the isosurfaces of trilinear functions is presented. Bas... more A characterization and classification of the isosurfaces of trilinear functions is presented. Based upon these results, a new algorithm for computing a triangular mesh approximation to isosurfaces for data given on a 3D rectilinear grid is presented. The original marching cubes algorithm is based upon linear interpolation along edges of the voxels. The asymptotic decider method is based upon bilinear interpolation on faces of the voxels. The algorithm of this paper carries this theme forward to using trilinear interpolation on the interior of voxels. The algorithm described here will produce a triangular mesh surface approximation to an isosurface which preserves the same connectivity/separation of vertices as given by the isosurface of trilinear interpolation.

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Cellular scaffolds in mammalian eggs

Frontiers in Bioscience, 2002

Introduction 3. Cellular scaffolds as a mechanism underlying cellular polarity and co-localizatio... more Introduction 3. Cellular scaffolds as a mechanism underlying cellular polarity and co-localization of factors 4. Conversion of the egg to the zygote: A brief overview 5. Signaling factors associated with structural elements in the egg/zygote 5.1. Calcium/calmodulin-dependent protein kinase II and the metaphase/anaphase transition 5.2. CaM KII and the midzone microtubules 5.3. Interaction between CaM KII and MAP Kinase 6. Conclusion 7. Acknowledgment 8. References

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GSK3β mediates acentromeric spindle stabilization by activated PKCζ

Developmental Biology, 2008

Upon fertilization, the mammalian egg undergoes a precise series of signaling events that orchest... more Upon fertilization, the mammalian egg undergoes a precise series of signaling events that orchestrate its conversion into a zygote. Mouse eggs contain acentrosomal spindle poles when arrested at meiotic metaphase II. The meiotic spindle is thought to provide a scaffold that mediates spatial and temporal regulation of the signaling pathways orchestrating post-fertilization events. Many kinases have been found to be enriched at the MII meiotic spindle, such as Protein Kinase C (PKC), and are thought to have an important role in regulating signaling events initiated through fertilization. In this study phosphorylated PKCζ (p-PKCζ) and Glycogen Synthase Kinase 3β (GSK3β) were found to be enriched at both acentrosomal spindle poles and the kinetochore region. Phosphorylated PKCζ (p-PKCζ) was immunopurified from MII eggs and was found to colocalize with known microtubule stabilizing components found in somatic cells, including GSK3β and Partition deficit protein 6 (Par6). Both fluorescence resonance energy transfer (FRET) and immunofluorescence confirmed the existence and close association of these proteins with p-PKCζ at the meiotic spindle. When GSK3β is phosphorylated on ser9 its activity is inhibited and the spindle is stabilized. However, when GSK3β is dephosphorylated (on ser9) it becomes active and the spindle is destabilized. The mechanism by which p-PKCζ maintains spindle organization appears to be through GSK3β and suggests that p-PKCζ phosphorylates GSK3β on the ser9 position inactivating GSK3β and consequently maintaining spindle stability during meiotic metaphase arrest.

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PKC isotypes in post-activated and fertilized mouse eggs: association with the meiotic spindle

Developmental Biology, 2004

Several isotypes of protein kinase C (PKC) have been reported to be expressed in mammalian eggs, ... more Several isotypes of protein kinase C (PKC) have been reported to be expressed in mammalian eggs, but it is unknown whether these isotypes have a common function in the egg during or within the first few hours of fertilization. Here we show that the isotypes of PKC exhibit distinct patterns of enrichment immediately after mouse egg activation. PKCa and g accumulate in the egg cortex 25 min postactivation, while only PKCa accumulates at the contractile ring of the forming second polar body about 1.5 h post-activation. PKC~exhibits some unique features that resulted in it being the focus of more extensive analysis. PKC~is tightly associated with the meiotic spindle as determined by detergent extraction and is closely associated with a-tubulin as determined by FRET analysis in the metaphase II (MII) egg. In addition, after egg activation, PKC~remains associated with the spindle as it transits into anaphase II and later telophase II, becoming associated with the midzone microtubules. Antibodies to the active form of PKC~are enriched on the spindle poles and later in development on the midzone microtubules. Active PKC~also is enriched in both pronuclei in the 6-h post-fertilization and in the 14-h post-fertilization embryo as well as in the nuclei of the two-cell embryo. Inhibition of PKC~, but not inhibition of other isotypes of PKC, results in rapid disruption of the meiotic spindle. This study suggests that PKC~has a role in spindle stability, while other PKC isotypes have different roles in the conversion of the egg to the zygote.

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Case Study: Comparing Two Methods for Filtering External Motion in 4D Confocal Microscopy Data

In this case study, we compare two methods for filtering external motion in time dependent volume... more In this case study, we compare two methods for filtering external motion in time dependent volume data sets acquired from confocal microscopy. The pros and cons of a landmark based and a voxel based method are discussed. We show that filtering external motion is an essential first step for the visualization of confocal data.

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The Role of Tyramine Within the Mouse Ovary

Microscopy and Microanalysis, 2019

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Methods for Creating and Evaluating 3D Tactile Images to Teach Stem Courses to the Visually Impaired

National Science Teachers Association, Jul 1, 2015