Dictyostelium Discoideum: Live Cell Imaging in Changing Perspective

2020 ◽  
pp. 126-145
Keyword(s):  
Dictyostelium Discoideum ◽  
Protein Localization ◽  
Red Light ◽  
Live Cell ◽  
Cell Organelle ◽  
Light Spectrum ◽  
Localization Study ◽  
Cell Localization ◽  
Uv Visible ◽  
Fluorescent Tags

The advent of advanced microscopes; during microscope evolution from simple microscopes to confocal and live cell microscope; having digital imaging facility revolutionized our view for the living cells. In the protein localization study, fluorescent proteins are tagged at amino or carboxyl (preferably) terminal of desired protein for live cell study. These live cell studies improved our understanding of protein dynamics and understanding its role in biological regulation. The mutational variants of fluorescent tags (GFP, RFP); can be used with different protein; which will efficiently use UV-Visible to Far Red light spectrum; without overlapping of excitation and emission spectrum. Further, various cell organelle (Lysosome, Golgi bodies, Endoplasmic Reticulum, Mitochondria, Nucleus) trackers; improved our live cell localization studies in the wide non-overlapping UV-Visible spectrum.This chapter gives an overview for live cell protein localization study in mitotically active, unicellular stage of Dictyostelium discoideum. This evolutionary cutting edge organism had both unicellular as well as multicellular stages during its life cycle. This chapter will provide the design of fusion of fluorescent tag to the specific gene and its live cell localization. Further, it will cover; transformation of the unicellular organism; drug based selection; sample preparation with nuclear, mitochondrial localization markers (trackers) and live cell localization study on live cell-confocal microscope setup. It will also have a glimpse of the design of fusion protein with an aspect of advantage and disadvantages.

scholarly journals The Preferences of Different Cultivars of Lettuce Seedlings (Lactuca sativa L.) for the Spectral Composition of Light

Agronomy ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1211
Author(s):  
Barbara Frąszczak ◽  
Monika Kula-Maximenko
Keyword(s):  
Chlorophyll Content ◽  
Blue Light ◽  
White Light ◽  
Leaf Shape ◽  
Red Light ◽  
Spectral Composition ◽  
Fresh Weight ◽  
Light Spectrum ◽  
Dark Green ◽  
Relative Chlorophyll Content

The spectrum of light significantly influences the growth of plants cultivated in closed systems. Five lettuce cultivars with different leaf colours were grown under white light (W, 170 μmol m−2 s−1) and under white light with the addition of red (W + R) or blue light (W + B) (230 μmol m−2 s−1). The plants were grown until they reached the seedling phase (30 days). Each cultivar reacted differently to the light spectrum applied. The red-leaved cultivar exhibited the strongest plasticity in response to the spectrum. The blue light stimulated the growth of the leaf surface in all the plants. The red light negatively influenced the length of leaves in the cultivars, but it positively affected their number in red and dark-green lettuce. It also increased the relative chlorophyll content and fresh weight gain in the cultivars containing anthocyanins. When the cultivars were grown under white light, they had longer leaves and higher value of the leaf shape index. The light-green cultivars had a greater fresh weight. Both the addition of blue and red light significantly increased the relative chlorophyll content in the dark-green cultivar. The spectrum enhanced with blue light had positive influence on most of the parameters under analysis in butter lettuce cultivars. These cultivars were also characterised by the highest absorbance of blue light.

scholarly journals Photocatalytic Activity and Characterization of Carbon-Modified Titania for Visible-Light-Active Photodegradation of Nitrogen Oxides

2012 ◽  
Vol 2012 ◽  
pp. 1-13 ◽  
Author(s):  
Chun-Hung Huang ◽  
Yu-Ming Lin ◽  
I-Kai Wang ◽  
Chun-Mei Lu
Keyword(s):  
Visible Light ◽  
Carbon Number ◽  
Red Light ◽  
Matrix Material ◽  
Carbon Sources ◽  
Impregnation Method ◽  
Light Spectrum ◽  
Precursor Molecule ◽  
Visible Light Active ◽  
Carbonate Species

A variety of carbon-modified titania powders were prepared by impregnation method using a commercial available titania powder, Hombikat UV100, as matrix material while a range of alcohols from propanol to hexanol were used as precursors of carbon sources. Rising the carbon number of alcoholic precursor molecule, the modified titania showed increasing visible activities ofNOxphotodegradation. The catalyst modified with cyclohexanol exhibited the best activities of 62%, 62%, 59%, and 54% for the totalNOxremoval under UV, blue, green, and red light irradiation, respectively. The high activity with long wavelength irradiation suggested a good capability of photocatalysis in full visible light spectrum. Analysis of UV-visible spectrum indicated that carbon modification promoted visible light absorption and red shift in band gap. XPS spectroscopic analysis identified the existence of carbonate species (C=O), which increased with the increasing carbon number of precursor molecule. Photoluminescence spectra demonstrated that the carbonate species suppressed the recombination rate of electron-hole pair. As a result, a mechanism of visible-light-active photocatalyst was proposed according to the formation of carbonate species on carbon-modified TiO2.

Dihydro-Si-Rhodamine for Live-Cell Localization Microscopy

2021 ◽  
Author(s):  
Xiaodong Zhang ◽  
Mengmeng Zhang ◽  
Yu Yan ◽  
Mingkang Wang ◽  
Jin Li ◽  
...  
Keyword(s):  
Cutting Edge ◽  
Live Cell ◽  
Fluorescence Properties ◽  
Localization Microscopy ◽  
Cell Localization

Fluorophores with photo-modulatory fluorescence properties are valuable for cutting-edge localization microscopy. The existing probes are either photo-activatable, or photo-switchable, but not both. We report a probe (DH-SiR), a leuco-dye obtained...

scholarly journals Live-cell nanoscopy enabled with transient labeling and the control of fluorophore blinking

2018 ◽  
Vol 190 ◽  
pp. 03008
Author(s):  
Alexander Mishin ◽  
Maxim Perfilov ◽  
Alexey Gavrikov ◽  
Anastasia Mamontova ◽  
Alexey Bogdanov ◽  
...  
Keyword(s):  
Super Resolution ◽  
Live Cell ◽  
Dynamic Replacement ◽  
Fluorescent Tags

Live-cell super-resolution of proteins labeled with genetically encoded fluorescent tags is a challenging task because of the imperfect labeling and the inevitable deterioration of the signal in the course of the experiment. Incomplete maturation of the covalently attached fluorescent tags, inefficient photoconversion, and photobleaching further complicate prolonged live-cell nanoscopy. We have implemented two strategies for lowering the photodamage: ensuring the dynamic replacement of damaged molecules and establishing conditions for the robust intrinsic blinking of the tags at lower illumination powers.

scholarly journals Photosynthetic Physiology of Blue, Green, and Red Light: Light Intensity Effects and Underlying Mechanisms

2021 ◽  
Vol 12 ◽  
Author(s):  
Jun Liu ◽  
Marc W. van Iersel
Keyword(s):  
Blue Light ◽  
Interactive Effect ◽  
Red Light ◽  
Green Light ◽  
Photosynthetic Photon Flux Density ◽  
Interactive Effects ◽  
Photon Flux ◽  
Photon Flux Density ◽  
Light Spectrum ◽  
Response Curves

Red and blue light are traditionally believed to have a higher quantum yield of CO2 assimilation (QY, moles of CO2 assimilated per mole of photons) than green light, because green light is absorbed less efficiently. However, because of its lower absorptance, green light can penetrate deeper and excite chlorophyll deeper in leaves. We hypothesized that, at high photosynthetic photon flux density (PPFD), green light may achieve higher QY and net CO2 assimilation rate (An) than red or blue light, because of its more uniform absorption throughtout leaves. To test the interactive effects of PPFD and light spectrum on photosynthesis, we measured leaf An of “Green Tower” lettuce (Lactuca sativa) under red, blue, and green light, and combinations of those at PPFDs from 30 to 1,300 μmol⋅m–2⋅s–1. The electron transport rates (J) and the maximum Rubisco carboxylation rate (Vc,max) at low (200 μmol⋅m–2⋅s–1) and high PPFD (1,000 μmol⋅m–2⋅s–1) were estimated from photosynthetic CO2 response curves. Both QYm,inc (maximum QY on incident PPFD basis) and J at low PPFD were higher under red light than under blue and green light. Factoring in light absorption, QYm,abs (the maximum QY on absorbed PPFD basis) under green and red light were both higher than under blue light, indicating that the low QYm,inc under green light was due to lower absorptance, while absorbed blue photons were used inherently least efficiently. At high PPFD, the QYinc [gross CO2 assimilation (Ag)/incident PPFD] and J under red and green light were similar, and higher than under blue light, confirming our hypothesis. Vc,max may not limit photosynthesis at a PPFD of 200 μmol m–2 s–1 and was largely unaffected by light spectrum at 1,000 μmol⋅m–2⋅s–1. Ag and J under different spectra were positively correlated, suggesting that the interactive effect between light spectrum and PPFD on photosynthesis was due to effects on J. No interaction between the three colors of light was detected. In summary, at low PPFD, green light had the lowest photosynthetic efficiency because of its low absorptance. Contrary, at high PPFD, QYinc under green light was among the highest, likely resulting from more uniform distribution of green light in leaves.

scholarly journals Photoselective Protective Netting Improves “Honeycrisp” Fruit Quality

Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1708
Author(s):  
Sara Serra ◽  
Stefano Borghi ◽  
Giverson Mupambi ◽  
Hector Camargo-Alvarez ◽  
Desmond Layne ◽  
...  
Keyword(s):  
Fruit Quality ◽  
Scattered Light ◽  
Red Light ◽  
Light Transmittance ◽  
Light Spectrum ◽  
Growing Seasons ◽  
Color Development ◽  
Red Color ◽  
Bitter Pit ◽  
And Control

High temperatures, wind, and excessive sunlight can negatively impact yield and fruit quality in semi-arid apple production regions. Netting was originally designed for hail protection, but it can modify the light spectrum and affect fruit quality. Here, pearl, blue, and red photoselective netting (≈20% shading factor) was installed in 2015 over a commercial “Cameron Select® Honeycrisp” orchard. Our research objectives were to (1) describe the light quantity and quality under the colored nets compared to an uncovered control and (2) investigate the effect of Photoselective nets on “Honeycrisp” apple quality for two growing seasons. Light transmittance and scattering for each treatment were measured with a spectroradiometer, and samples for fruit quality analyses were collected at harvest. PAR (photosynthetic active radiation), UV, blue, red, and far-red light were lower underneath all netting treatments compared to an uncovered control. The scattered light was higher under the pearl net compared to other colors, while red and far-red light were lower under the blue net. For two consecutive years, trees grown under the photoselective nets intercepted more incoming light than the uncovered trees with no differences among the three colors. In both years, trees under red and blue nets had more sunburn-free (clean) apples than pearl and control. Red color development for fruit was lower when nets were used. Interestingly, bitter pit incidence was lower underneath red nets for both years. Other than red color development, “Honeycrisp” fruit quality was not appreciably affected by the use of netting. These results highlight the beneficial effect of nets in improving light quality in orchards and mitigating physiological disorders such as bitter pit in “Honeycrisp” apple.

scholarly journals Stressful colours: corticosterone concentrations in a free-living songbird vary with the spectral composition of experimental illumination

2015 ◽  
Vol 11 (8) ◽  
pp. 20150517 ◽  
Author(s):  
Jenny Q. Ouyang ◽  
Maaike de Jong ◽  
Michaela Hau ◽  
Marcel E. Visser ◽  
Roy H. A. van Grunsven ◽  
...  
Keyword(s):  
Large Scale ◽  
Red Light ◽  
Spectral Composition ◽  
Parus Major ◽  
Light Spectrum ◽  
Night Time ◽  
Free Living ◽  
Treatment Type ◽  
Artificial Illumination ◽  
Living Organisms

Organisms have evolved under natural daily light/dark cycles for millions of years. These cycles have been disturbed as night-time darkness is increasingly replaced by artificial illumination. Investigating the physiological consequences of free-living organisms in artificially lit environments is crucial to determine whether nocturnal lighting disrupts circadian rhythms, changes behaviour, reduces fitness and ultimately affects population numbers. We make use of a unique, large-scale network of replicated field sites which were experimentally illuminated at night using lampposts emanating either red, green, white or no light to test effect on stress hormone concentrations (corticosterone) in a songbird, the great tit ( Parus major ). Adults nesting in white-light transects had higher corticosterone concentrations than in the other treatments. We also found a significant interaction between distance to the closest lamppost and treatment type: individuals in red light had higher corticosterone levels when they nested closer to the lamppost than individuals nesting farther away, a decline not observed in the green or dark treatment. Individuals with high corticosterone levels had fewer fledglings, irrespective of treatment. These results show that artificial light can induce changes in individual hormonal phenotype. As these effects vary considerably with light spectrum, it opens the possibility to mitigate these effects by selecting street lighting of specific spectra.

Syntheses of lactosyl ceramide analogues carrying novel bifunctional BODIPY dyes directed towards the differential analysis of multiplexed glycosphingolipids by MS/MS using iTRAQ

2014 ◽  
Vol 50 (23) ◽  
pp. 3010-3013 ◽  
Author(s):  
Sang-Hyun Son ◽  
Shusaku Daikoku ◽  
Atsuko Ohtake ◽  
Katsuhiko Suzuki ◽  
Kazuya Kabayama ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Differential Analysis ◽  
Fluorescent Tags

Lactosyl ceramide analogues carrying novel bifunctional BODIPY-based fluorescent tags were designed and synthesised for live cell imaging.

scholarly journals Investigation of Growth Rate and Phycocolloid Content from Kappaphycus alvarezii (Rhodophyta, Solieriaceae) under Different Light Conditions Using Vibrational Spectroscopy

2016 ◽  
Vol 4 (2) ◽  
pp. 1 ◽  
Author(s):  
Vun Yee Thien ◽  
Kenneth Francis Rodrigues ◽  
Clemente Michael Vui Ling Wong ◽  
Wilson Thau Lym Yong
Keyword(s):  
Carbon Dioxide ◽  
Growth Rate ◽  
Chemical Structure ◽  
Kappaphycus Alvarezii ◽  
Red Light ◽  
Monochromatic Light ◽  
Carbon Dioxide Concentration ◽  
Major Constituent ◽  
Light Spectrum ◽  
Spectral Profiles

<p><em>Kappa</em>-carrageenan (<em>K</em>-carrageenan) is an important phycocolloid which is a major constituent of the cell wall of <em>Kappaphycus alvarezii</em>. The chemical structure of <em>K</em>-carrageenan comprises a linear backbone of D-galactose residues linked with alternating α-(1,3) and β-(1,4) linkages which are substituted by one ester-sulphonic group per di-galactose repeating unit. The spectral qualities of light as well as the ambient carbon dioxide concentration, both play an important role in the photosynthetic pathway in plants and this investigation set forth to establish the effect of different wavelengths of light and carbon dioxide supplementation on the chemical structure of <em>K</em>-carrageenan obtained from <em>K. alvarezii. </em>Specimens were cultivated under a range of monochromatic light spectra and assessed for chemical composition using Fourier Transform Infrared (FTIR) spectroscopy.The <em>K. alvarezii</em> control was irradiated with full light spectrum, treatments were carried out using blue (492-455 nm), green (577-492 nm) and red (780-622 nm)light. One experiment was carried out by supplementation with carbon dioxide. Samples were collected after 14 days. The effect of different wavelengths of light on the growth rates of experimental samples was determined. Red light had the most significant impact on the growth rate of <em>K. alvarezii </em>as compared to those treated with blue light. The FTIR fingerprint of the ground seaweed was found to be identical to that of commercial <em>K</em>-carrageenan (Sigma). Special emphasis was given to the 800-1300 cm<sup>-1 </sup>region, which presents several vibrational modes. All the samples produced similar FTIR spectral profiles, suggesting that genes related to the carrageenan biosynthesis are not affected by different wavelengths of light or CO<sub>2</sub>. The results obtained from FTIR spectroscopy demonstrated that different wavelengths of light and supplementation with CO<sub>2</sub> have no influence to the chemical structure of <em>K</em>-carrageenan in <em>K. alvarezii</em>.</p>

scholarly journals Live‐Cell Localization Microscopy with a Fluorogenic and Self‐Blinking Tetrazine Probe

2020 ◽  
Vol 59 (2) ◽  
pp. 804-810 ◽  
Author(s):  
Philipp Werther ◽  
Klaus Yserentant ◽  
Felix Braun ◽  
Nicolai Kaltwasser ◽  
Christoph Popp ◽  
...  
Keyword(s):  
Live Cell ◽  
Localization Microscopy ◽  
Cell Localization
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