Thousand-fold Increase in Plasmonic Light Emission via Combined Electronic and Optical Excitations

ABSTRACTWe describe the acquisition of Raman and photoluminescence (PL) spectra on porous silicon (PS) samples that emit visible light. Spectra were acquired in both ex situ experiments (after exposure to air) and in situ experiments (with the PS covered either with the hydrofluoric acid electrolyte used in the formation process or water). Our results generally show a correlation of blue-shifted PL with increased oxidation. In one set of ex situ experiments, however, we observed an inconsistency in the shift of the wavelengthof maximum luminescence intensity for PS samples that exhibit oxygenated character in the Raman spectra. A higher anodization current density produced a red shift in the PL spectra in one experiment, while chemical dissolution of the PS by hydrofluoric acid produced the well-known blue shift in the other case. In two in situ experiments, we observed very weak and red-shifted PL for a PS sample immersed in HF (compared to the same sample measured later in air) while in another we immersed air-exposed PS in water and observed a 15-fold increase in PL intensity along with a blue shift in the luminescence maximum.

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Measuring C. elegans spatial foraging and food intake using bioluminescent bacteria

10.1101/759928 ◽

2019 ◽

Cited By ~ 1

Author(s):

Siyu Serena Ding ◽

Karen S. Sarkisyan ◽

Andre E. X. Brown

Keyword(s):

Food Intake ◽

Foraging Behaviour ◽

Light Emission ◽

Fold Increase ◽

Food Patch ◽

Background Signal ◽

Bioluminescent Bacteria ◽

C Elegans ◽

Tightly Coupled ◽

Lower Background

ABSTRACTFor most animals, feeding includes two behaviours: foraging to find a food patch and food intake once a patch is found. The nematode Caenorhabditis elegans is a useful model for studying the genetics of both behaviours. However, most methods of measuring feeding in worms quantify either foraging behaviour or food intake but not both. Imaging the depletion of fluorescently labelled bacteria provides information on both the distribution and amount of consumption, but even after patch exhaustion a prominent background signal remains, which complicates quantification. Here, we used a bioluminescent Escherichia coli strain to quantify C. elegans feeding. With light emission tightly coupled to active metabolism, only living bacteria are capable of bioluminescence so the signal is lost upon ingestion. We quantified the loss of bioluminescence using N2 reference worms and eat-2 mutants, and found a nearly 100-fold increase in signal-to-background ratio and lower background compared to loss of fluorescence. We also quantified feeding using aggregating npr-1 mutant worms. We found that groups of npr-1 mutants first clear bacteria from each other before foraging collectively for more food; similarly, during high density swarming, only worms at the migrating front are in contact with bacteria. These results demonstrate the usefulness of bioluminescent bacteria for quantifying feeding and suggest a hygiene hypothesis for the function of C. elegans aggregation and swarming.

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Measuring Caenorhabditis elegans Spatial Foraging and Food Intake Using Bioluminescent Bacteria

Genetics ◽

10.1534/genetics.119.302804 ◽

2020 ◽

Vol 214 (3) ◽

pp. 577-587 ◽

Cited By ~ 3

Author(s):

Siyu Serena Ding ◽

Maksym Romenskyy ◽

Karen S. Sarkisyan ◽

Andre E. X. Brown

Keyword(s):

Caenorhabditis Elegans ◽

Food Intake ◽

Light Emission ◽

Large Population ◽

Fold Increase ◽

Bioluminescent Bacteria ◽

C Elegans ◽

Link Type ◽

Tightly Coupled ◽

Lower Background

For most animals, feeding includes two behaviors: foraging to find a food patch and food intake once a patch is found. The nematode Caenorhabditis elegans is a useful model for studying the genetics of both behaviors. However, most methods of measuring feeding in worms quantify either foraging behavior or food intake, but not both. Imaging the depletion of fluorescently labeled bacteria provides information on both the distribution and amount of consumption, but even after patch exhaustion a prominent background signal remains, which complicates quantification. Here, we used a bioluminescent Escherichia coli strain to quantify C. elegans feeding. With light emission tightly coupled to active metabolism, only living bacteria are capable of bioluminescence, so the signal is lost upon ingestion. We quantified the loss of bioluminescence using N2 reference worms and eat-2 mutants, and found a nearly 100-fold increase in signal-to-background ratio and lower background compared to loss of fluorescence. We also quantified feeding using aggregating npr-1 mutant worms. We found that groups of npr-1 mutants first clear bacteria from within the cluster before foraging collectively for more food; similarly, during large population swarming, only worms at the migrating front are in contact with bacteria. These results demonstrate the usefulness of bioluminescent bacteria for quantifying feeding and generating insights into the spatial pattern of food consumption.

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Bacterial luciferase alpha beta fusion protein is fully active as a monomer and highly sensitive in vivo to elevated temperature

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.86.17.6528 ◽

1989 ◽

Vol 86 (17) ◽

pp. 6528-6532 ◽

Cited By ~ 61

Author(s):

A Escher ◽

D J O'Kane ◽

J Lee ◽

A A Szalay

Keyword(s):

Elevated Temperature ◽

Fusion Protein ◽

Transcriptional Control ◽

Light Emission ◽

Fold Increase ◽

Wild Type ◽

Bacterial Luciferase ◽

T7 Promoter ◽

Alpha Beta

A 2.2-kilobase-pair (kbp) DNA fragment from Vibrio harveyi contains the luxA and luxB genes separated by a 26-base-pair (bp) intergenic region. The two genes were converted to a single open reading frame by site-specific mutagenesis. A full-length fusion protein is obtained when the new gene is placed under transcriptional control of a T7 promoter in Escherichia coli. Bioluminescence of colonies containing the gene fusion is 0.002% of the wild-type luciferase [alkanal monooxygenase (FMN-linked); alkanal, reduced-FMN:oxygen oxidoreductase (1-hydroxylating, luminescing), EC 1.14.14.3] at 37 degrees C. Growth at 23 degrees C results in a greater than 50,000-fold increase in light emission in cells containing fusion protein, whereas only a 3-fold increase in observed with cells containing the luxAB dicistron. Purified fusion protein isolated from E. coli grown at 19 degrees C exists in both monomeric and dimeric forms with specific bioluminescence activities comparable to the heterodimeric wild-type enzyme at 23 degrees C and 37 degrees C. These findings show that the alpha beta fusion polypeptide is functional as a monomer and suggest that its folding is drastically affected at elevated temperature. We hypothesize that the two-subunit bacterial luciferase may have evolved from a monomer as a result of a temperature increase in the environment.

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Renilla Bioluminescence: A Morphologic Approach to the Location of Photocytes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100051050 ◽

1974 ◽

Vol 32 ◽

pp. 208-209

Author(s):

Ben O. Spurlock ◽

Milton J. Cormier

Keyword(s):

Excited State ◽

Ground State ◽

Molecular Basis ◽

Light Emission ◽

Chemical Basis ◽

Electronic Excited State ◽

Colonial Form ◽

The Common ◽

Eighteenth And Nineteenth Centuries ◽

Catalyzed Oxidation

The phenomenon of bioluminescence has fascinated layman and scientist alike for many centuries. During the eighteenth and nineteenth centuries a number of observations were reported on the physiology of bioluminescence in Renilla, the common sea pansy. More recently biochemists have directed their attention to the molecular basis of luminosity in this colonial form. These studies have centered primarily on defining the chemical basis for bioluminescence and its control. It is now established that bioluminescence in Renilla arises due to the luciferase-catalyzed oxidation of luciferin. This results in the creation of a product (oxyluciferin) in an electronic excited state. The transition of oxyluciferin from its excited state to the ground state leads to light emission.

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Detection of viruses by electron microscopy: Increased sensitivity by direct micro-ultracentrifugation of samples

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100128821 ◽

1987 ◽

Vol 45 ◽

pp. 908-909

Author(s):

Alain R. Trudel ◽

M. Trudel

Keyword(s):

Electron Microscopy ◽

Fold Increase ◽

Observation Time ◽

Clinical Samples ◽

Maximum Speed ◽

Viral Particles ◽

Structural Details ◽

Latex Spheres ◽

Increased Sensitivity ◽

Size Of Particles

AirfugeR (Beckman) direct ultracentrifugation of viral samples on electron microscopy grids offers a rapid way to concentrate viral particles or subunits and facilitate their detection and study. Using the A-100 fixed angle rotor (30°) with a K factor of 19 at maximum speed (95 000 rpm), samples up to 240 μl can be prepared for electron microscopy observation in a few minutes: observation time is decreased and structural details are highlighted. Using latex spheres to calculate the increase in sensitivity compared to the inverted drop procedure, we obtained a 10 to 40 fold increase in sensitivity depending on the size of particles. This technique also permits quantification of viral particles in samples if an aliquot is mixed with latex spheres of known concentration.Direct ultracentrifugation for electron microscopy can be performed on laboratory samples such as gradient or column fractions, infected cell supernatant, or on clinical samples such as urine, tears, cephalo-rachidian liquid, etc..

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Advantages of Complementary Stereo Images as Viewed with the Low-Temperature Field-Emission SEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100131206 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1314-1315

Author(s):

William P. Wergin ◽

Eric F. Erbe

Keyword(s):

Fold Increase ◽

Horizontal Displacement ◽

Three Dimensions ◽

Stereo Image ◽

Stereo Pair ◽

Sem Micrograph ◽

Left And Right ◽

The Common ◽

The Brain ◽

Pair Analysis

The eye-brain complex allows those of us with normal vision to perceive and evaluate our surroundings in three-dimensions (3-D). The principle factor that makes this possible is parallax - the horizontal displacement of objects that results from the independent views that the left and right eyes detect and simultaneously transmit to the brain for superimposition. The common SEM micrograph is a 2-D representation of a 3-D specimen. Depriving the brain of the 3-D view can lead to erroneous conclusions about the relative sizes, positions and convergence of structures within a specimen. In addition, Walter has suggested that the stereo image contains information equivalent to a two-fold increase in magnification over that found in a 2-D image. Because of these factors, stereo pair analysis should be routinely employed when studying specimens.Imaging complementary faces of a fractured specimen is a second method by which the topography of a specimen can be more accurately evaluated.

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Scanning luminescence x-ray microscopy: Progress toward selective staining using microspheres

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100168104 ◽

1994 ◽

Vol 52 ◽

pp. 74-75

Author(s):

C. Jacobsen ◽

J. Fu ◽

S. Mayer ◽

Y. Wang ◽

S. Williams

Keyword(s):

Visible Light ◽

Active Sites ◽

Light Emission ◽

Chinese Hamster ◽

Polystyrene Spheres ◽

Good Resistance ◽

X Ray ◽

Selective Staining ◽

Visible Light Emission ◽

Specific Labelling

In scanning luminescence x-ray microscopy (SLXM), a high resolution x-ray probe is used to excite visible light emission (see Figs. 1 and 2). The technique has been developed with a goal of localizing dye-tagged biochemically active sites and structures at 50 nm resolution in thick, hydrated biological specimens. Following our initial efforts, Moronne et al. have begun to develop probes based on biotinylated terbium; we report here our progress towards using microspheres for tagging.Our initial experiments with microspheres were based on commercially-available carboxyl latex spheres which emitted ~ 5 visible light photons per x-ray absorbed, and which showed good resistance to bleaching under x-ray irradiation. Other work (such as that by Guo et al.) has shown that such spheres can be used for a variety of specific labelling applications. Our first efforts have been aimed at labelling ƒ actin in Chinese hamster ovarian (CHO) cells. By using a detergent/fixative protocol to load spheres into cells with permeabilized membranes and preserved morphology, we have succeeded in using commercial dye-loaded, spreptavidin-coated 0.03μm polystyrene spheres linked to biotin phalloidon to label f actin (see Fig. 3).

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The Advantages of Cryotechniques: Application To Bioluminescent Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010018118x ◽

1990 ◽

Vol 48 (1) ◽

pp. 486-487

Author(s):

Gisèle Nicolas ◽

Jean-Marie Bassot ◽

Marie-Thérèse Nicolas

Keyword(s):

Endoplasmic Reticulum ◽

Striated Muscle ◽

Light Emission ◽

Stable State ◽

Freeze Substitution ◽

Initial Step ◽

Point Of Interest ◽

Cell Components ◽

Excellent Preservation ◽

External Water

The use of fast-freeze fixation (FFF) followed by freeze-substitution (FS) brings substantial advantages which are due to the extreme rapidity of this fixation compared to the conventional one. The initial step, FFF, physically immobilizes most molecules and therefore arrests the biological reactions in a matter of milliseconds. The second step, FS, slowly removes the water content still in solid state and, at the same time, chemically fixes the other cell components in absence of external water. This procedure results in an excellent preservation of the ultrastructure, avoids osmotic artifacts,maintains in situ most soluble substances and keeps up a number of cell activities including antigenicities. Another point of interest is that the rapidity of the initial immobilization enables the capture of unstable structures which, otherwise, would slip towards a more stable state. When combined with electrophysiology, this technique arrests the ultrastructural modifications at a well defined state, allowing a precise timing of the events.We studied the epithelium of the elytra of the scale-worm, Harmothoe lunulata which has excitable, conductible and bioluminescent properties. The intracellular sites of the light emission are paracrystals of endoplasmic reticulum (PER), named photosomes (Fig.1). They are able to flash only when they are coupled with plasma membrane infoldings by dyadic or triadic junctions (Fig.2) basically similar to those of the striated muscle fibers. We have studied them before, during and after stimulation. FFF-FS showed that these complexes are labile structures able to diffentiate and dedifferentiate within milliseconds. Moreover, a transient network of endoplasmic reticulum was captured which we have named intermediate endoplasmic reticulum (IER) surrounding the PER (Fig.1). Numerous gap junctions are found in the membranous infoldings of the junctional complexes (Fig.3). When cryofractured, they cleave unusually (Fig.4-5). It is tempting to suggest that they play an important role in the conduction of the excitation.

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Prevalence of Self-Reported Depression Symptoms and Perceived Anxiety Among Community-Dwelling U.S. Adults Reporting Tinnitus

Perspectives of the ASHA Special Interest Groups ◽

10.1044/2020_persp-19-00178 ◽

2020 ◽

Vol 5 (4) ◽

pp. 959-970

Author(s):

Kelly M. Reavis ◽

James A. Henry ◽

Lynn M. Marshall ◽

Kathleen F. Carlson

Keyword(s):

Mental Health ◽

Fold Increase ◽

Community Dwelling ◽

Self Report ◽

Depression Symptoms ◽

Community Based Interventions ◽

Data Set ◽

Health Distress ◽

Validated Questionnaire ◽

Mental Health Distress

Purpose The aim of this study was to examine the relationship between tinnitus and self-reported mental health distress, namely, depression symptoms and perceived anxiety, in adults who participated in the National Health and Nutrition Examinations Survey between 2009 and 2012. A secondary aim was to determine if a history of serving in the military modified the associations between tinnitus and mental health distress. Method This was a cross-sectional study design of a national data set that included 5,550 U.S. community-dwelling adults ages 20 years and older, 12.7% of whom were military Veterans. Bivariable and multivariable logistic regression was used to estimate the association between tinnitus and mental health distress. All measures were based on self-report. Tinnitus and perceived anxiety were each assessed using a single question. Depression symptoms were assessed using the Patient Health Questionnaire, a validated questionnaire. Multivariable regression models were adjusted for key demographic and health factors, including self-reported hearing ability. Results Prevalence of tinnitus was 15%. Compared to adults without tinnitus, adults with tinnitus had a 1.8-fold increase in depression symptoms and a 1.5-fold increase in perceived anxiety after adjusting for potential confounders. Military Veteran status did not modify these observed associations. Conclusions Findings revealed an association between tinnitus and both depression symptoms and perceived anxiety, independent of potential confounders, among both Veterans and non-Veterans. These results suggest, on a population level, that individuals with tinnitus have a greater burden of perceived mental health distress and may benefit from interdisciplinary health care, self-help, and community-based interventions. Supplemental Material https://doi.org/10.23641/asha.12568475

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