Expansion microscopy Chen, Fei; Tillberg, Paul W.; Boyden, Edward S.
Science (American Association for the Advancement of Science),
01/2015, Letnik:
347, Številka:
6221
Journal Article
Recenzirano
Odprti dostop
In optical microscopy, fine structural details are resolved by using refraction to magnify images of a specimen. We discovered that by synthesizing a swellable polymer network within a specimen, it ...can be physically expanded, resulting in physical magnification. By covalently anchoring specific labels located within the specimen directly to the polymer network, labels spaced closer than the optical diffraction limit can be isotropically separated and optically resolved, a process we call expansion microscopy (ExM). Thus, this process can be used to perform scalable superresolution microscopy with diffraction-limited microscopes. We demonstrate ExM with apparent ∼70-nanometer lateral resolution in both cultured cells and brain tissue, performing three-color superresolution imaging of ∼107 cubic micrometers of the mouse hippocampus with a conventional confocal microscope.
Mikroskop cahaya dapat digunakan untuk melakukan magnifikasi dan melihat objek mikroskopis seperti bakteri, virus, atau sel untuk tujuan diagnosis suatu penyakit. Dalam melakukan pengamatan ...berdasarkan jumlah sampel yang banyak, ahli laboratorium rentan terhadap kelelahan, kesalahan, dan subjektivitas pengamatan. Oleh karena itu, kamera digital, komputer mini raspberry pi, layar liquid crystal display (LCD), motor stepper, catu daya dan bahasa pemrograman python digunakan sebagai komponen pendukung mikroskop cahaya untuk mengakusisi sampel laboratorium yang diamati dalam format digital sehingga dapat dilakukan operasi pengolahan citra pada raspberry pi dan ditampilkan hasilnya di layar LCD. Sedangkan motor stepper dan driver motor digunakan untuk menggeser meja sampel secara otomatis. Semua proses ini dilakukan dengan menggunakan bahasa pemrograman python. Hal ini dilakukan dengan tujuan untuk mengurangi beban pengamat dalam melihat dan menggeser slide sampel sebanyak 100 kali. Sistem akuisisi citra digital dan penggeseran meja sampel otomatis ini dirancang pada mikroskop cahaya portabel mampu meng-capture sampel pada kaca preparat menjadi citra digital, menggeser kaca preparat 100 kali, dan dilengkapi dengan catu daya 220 Volt sehingga dapat digunakan di fasilitas kesehatan manapun di Indonesia. Mikroskop digital, otomatis, dan portabel berbasis platform komputer mini raspberry pi ini dapat digunakan untuk melakukan pemeriksaan laboratorium berbagai penyakit seperti tuberculosis, malaria, atau leukimia dengan efisien dan efektif sehingga dapat mempermudah proses diagnosis penyakit dan dapat meningkatkan layanan kesehatan di berbagai daerah di Indonesia dan berkontribusi dalam penurunan dan eliminasi berbagai penyakit.
The Zeiss Airyscan microscope transforms a diffraction-limited, point-scanning confocal microscope into a super-resolution microscope using a specialized 32-channel Airyscan detector. By improving ...resolution twofold and signal-to-noise ratio eightfold relative to conventional confocal microscopes while retaining confocal functionality, the Airyscan microscope has become a very popular super-resolution imaging tool for cell biologists. In this chapter, we describe the fundamentals of Airyscan imaging, with the aim of helping the reader determine the proper acquisition settings for different types of experiments, optimize imaging conditions, and process the raw Airyscan images to obtain final images with the best quality. We also provide some tips, tricks, and best practices for Airyscan imaging. Of note, while our focus is on the Airyscan function of this microscope rather than its conventional confocal function, the Airyscan unit comes as an add-on to the conventional Zeiss laser scanning confocal microscope. This protocol is for the first generation Airyscan Zeiss 800 series microscope.
This review commemorates the 40th anniversary of DNA sequencing, a period in which we have already witnessed multiple technological revolutions and a growth in scale from a few kilobases to the first ...human genome, and now to millions of human and a myriad of other genomes. DNA sequencing has been extensively and creatively repurposed, including as a 'counter' for a vast range of molecular phenomena. We predict that in the long view of history, the impact of DNA sequencing will be on a par with that of the microscope.
The nanoscale control afforded by scanning probe microscopes has prompted the development of a wide variety of scanning-probe-based patterning methods. Some of these methods have demonstrated a high ...degree of robustness and patterning capabilities that are unmatched by other lithographic techniques. However, the limited throughput of scanning probe lithography has prevented its exploitation in technological applications. Here, we review the fundamentals of scanning probe lithography and its use in materials science and nanotechnology. We focus on robust methods, such as those based on thermal effects, chemical reactions and voltage-induced processes, that demonstrate a potential for applications.
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•Superhydrophobic POSS-GO was obtained by grafting reaction of POSS-NH2 and GO.•The dispersibility of GO was greatly improved by connecting POSS in many solvents.•The POSS-GO with ...special function was good for anticorrosion ability of coating.
Super-hydrophobicity polyhedral oligomeric silsesquioxane-modified graphene oxide (POSS-GO) was synthesized by one-step reaction between graphene oxide (GO) and aminopropylisobutyl polyhedral oligomeric silsesquioxane (POSS-NH2). Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) spectra indicated that the POSS were successfully connected to the edge and surface of GO nanosheets. Scanning probe microscope (SPM) and transmission electron microscope (TEM) images demonstrated that the POSS-GO with a thickness of 1.58 nm presented a stably dispersion in anhydrous ethanol. Results from electrochemical behaviors showed that the proper incorporation of POSS-GO could availably improve the anticorrosion ability of epoxy based coatings in simulate marine environment (3.5 wt% NaCl solution). The good dispersion of POSS-GO helped to fill the original defect and enhanced the complication of penetration path. The super-hydrophobicity of POSS-GO was beneficial to the reduction of actual contact zone. The synergistic effect of good dispersion and super-hydrophobicity promoted the enhancement of anticorrosion ability for composite coatings.
Principle has it that even the most advanced super‐resolution microscope would be futile in providing biological insight into subcellular matrices without well‐designed fluorescent tags/probes. ...Developments in biology have increasingly been boosted by advances of chemistry, with one prominent example being small‐molecule fluorescent probes that not only allow cellular‐level imaging, but also subcellular imaging. A majority, if not all, of the chemical/biological events take place inside cellular organelles, and researchers have been shifting their attention towards these substructures with the help of fluorescence techniques. This Review summarizes the existing fluorescent probes that target chemical/biological events within a single organelle. More importantly, organelle‐anchoring strategies are described and emphasized to inspire the design of new generations of fluorescent probes, before concluding with future prospects on the possible further development of chemical biology.
See below the surface: Fluorescent probes that target individual organelles and elucidate their functionalities are systematically summarized in this Review. The design strategy towards organelle targeting will shed light on basic studies of cell biology.
There is limited literature on the modulation of fat crystallization by natural waxes, which are underutilized and cost-effective food industry resources. In this study, the effects of two natural ...waxes (beeswax (BW) and carnauba wax (CW)) on the crystallization behavior of palm kernel stearin (PKS85) were systematically investigated by using differential scanning calorimetry (DSC), pulsed nuclear magnetic resonance (p-NMR), X-ray diffraction (XRD) and polarized light microscope (PLM). CW addition significantly promoted the isothermal crystallization process of PKS85, especially at 20 °C, while BW had no obvious effect on the crystallization rate. BW and CW addition changed the crystal growth mode at 20 °C. BW and CW could participate in the crystallization of TAGs in PKS85, induce the formation of a new hydrocarbon chain distances of 3.70 and 4.15 Å and decrease the lamellar distance (d(001)) and domain size (ξ) of the single crystallites, leading to a different morphology of fat crystals. Our findings indicated that BW and CW could modulate the fat crystal structure and possibly engineer the functional properties of fats and fat-structured materials, which also provided new application of natural waxes in the food industry.
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•CW addition promoted the isothermal crystallization process of PKS85.•BW and CW addition changed the crystal growth mode at 20 °C.•BW and CW induced a new hydrocarbon chain distances of 3.70 and 4.15 Å.•BW and CW decreased the lamellar distance and domain size of single crystallite.•BW and CW significantly changed the crystal morphology of PKS85.
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•Sewage Treatment Plants are significant sources of microplastics (MPs).•Up to 99% of MPs’ can be removed during conventional wastewater (WW) treatment.•MPs accumulate to sludge, lack ...of data for possible changes during treatment.•Careful selection of advanced treatment process has positive impact on MPs removal.•A standardized protocol for MPs’ sampling, pretreatment and analysis is needed.
Microplastics are plastic fragments lower than 5 mm that are detected in the environment causing various effects on organisms. Several research articles have recognized Sewage Treatment Plants as important sources of polyethylene and polypropylene beads, polyester, polyamide and other types of microplastics. For their determination, techniques such as visual identification using microscope, Fourier-transform infrared and RAMAN spectroscopy are used, while chemical oxidation, enzymatic maceration and density separation are applied as pretreatment methods for the removal of the inorganic and organic content. Microplastics’ concentrations range up to 3160 particles L−1, 125 particles L−1 and 170.9 × 103 particles Kg-1 TS dw in raw, treated wastewater and sludge, respectively. Their removal during wastewater treatment ranges between 72% and 99.4%; the main processes that contribute to their removal are primary and secondary treatment, while the effect of tertiary treatment depends on the applied technology. Entrapment in suspended solids and accumulation to sludge are the major mechanisms governing their fate. A standardized protocol for samples’ collection and pretreatment as well as microplastics’ isolation and characterization is needed; future reseach should investigate the possible chemical and physical changes of microplastics during treatment, and their role as carriers for the transfer of emerging micropollutants.
Imaging the transcriptome in situ with high accuracy has been a major challenge in single-cell biology, which is particularly hindered by the limits of optical resolution and the density of ...transcripts in single cells
. Here we demonstrate an evolution of sequential fluorescence in situ hybridization (seqFISH+). We show that seqFISH+ can image mRNAs for 10,000 genes in single cells-with high accuracy and sub-diffraction-limit resolution-in the cortex, subventricular zone and olfactory bulb of mouse brain, using a standard confocal microscope. The transcriptome-level profiling of seqFISH+ allows unbiased identification of cell classes and their spatial organization in tissues. In addition, seqFISH+ reveals subcellular mRNA localization patterns in cells and ligand-receptor pairs across neighbouring cells. This technology demonstrates the ability to generate spatial cell atlases and to perform discovery-driven studies of biological processes in situ.
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