Introduction to women in microscopy: Volume 2 DOI
Michelle Peckham, Ulla Neumann, S J Culley

et al.

Journal of Microscopy, Journal Year: 2024, Volume and Issue: 295(1), P. 3 - 5

Published: June 11, 2024

As with the first volume of 'Women in Microscopy', we have assembled a wide-ranging collection papers focus on microscopy techniques and technologies encompassing five contributions topics as diverse surface passivation functionalisation for mass photometry applications, innovative time-modulated structured illumination single-molecule localisation microscopy, Mesolens imaging large fields view at low magnification yet high resolution, array tomography mainly but not exclusively 3D electron quantification life sciences microscopy. We would like to thank once more all authors two fantastic volumes Microscopy' their participation. The return rate after our initial call demonstrated interest this topic throughout community. second starts letter editor implementation an interdisciplinary bootcamp UC Berkeley intended promote supportive networking women alleviate gender-related inequities field microscopy.1 In early 1990s, when Ulla Neumann was undergraduate student, she asked practical course by male instructor German university if knew what photomultiplier was—at time didn't. And pointed out that kind information could evidently be found Brigitte, typical 'women's magazine'. Meagan Esbin Lena Blackman shows some progress has been made over last three decades. Still, given global only about 10% are late-stage careers, is excellent example building network focusing incoming students STEM. Through lectures, demos, nanorotations industry tours, opportunity connect role models different, or less public/private settings both academia industry, providing academic also career development support. future, specifically include trans gender nonconforming diversity initiatives. technical note Suliana Manley's group describes improvements glass coatings cover slips reduce nonspecific binding label-free, single-particle based interferometric scattering (iSCAT) microscopy.2 This technique relies interference between light scattered single particles reflected coverslip. When observing weakly scattering, proteins, detection signal above background noise caused impurities remains challenge. While various routinely used fluorescence studies, case photometry. described note, successfully PEG molecular weight (2 kDa) combination APTES (3-aminopropyltriethoxysilane), producing high-quality (smaller dynamic signal) evidenced equal numbers detected unbinding events different test proteins. Surface functionalisation, process which target molecules displayed surface, efficient using PEG-maleimide create covalent thioether bonds thiol groups comparison positively charging surfaces poly-L-lysine. contribution will useful applications other label-free techniques. One perpetual quest localise individual ever-increasing precision, bypassing diffraction limit light. work Sandrine Lévêque-Fort's temporal method changes improve precision.3 random subset fluorescent specific emitting excitation ('blinking') allows spatially analyse its point spread function (based models) calculate localisation. Acquisition thousands images, each recording molecules, reconstruct emitters entire analysed. paper presents approach shifted temporarily either xy xz resulting sinusoid, temporally modulated emission fluorophores depending position positional being carried phase signal. Key novel named ModLoc (Modulated Localisation) strategy acquire four intensity values necessary extract demodulation within same camera frame frequency, combined use digital micromirrors devices modulation galvanometric mirrors optical setup. With help combination, achieve twofold improvement precision conventional approaches enabling higher spatial excitation, adaption objective parameters possibility extend tracking studies. Gail McConnell's introduces us into world mesoscale standing wave imaging, combining axially (a sinusoidal pattern generated interfering beams) image formation through low-magnification, high-numerical aperture lens produce high-resolution images fluorescently labelled, three-dimensional structures 2D dataset.4 Thus far, so-called (NA 0.47, 4×), McConnell colleagues 2016,5 number modalities (confocal, sheet, colour bright-field, wide-field epifluorescence, FLIM). However, used, one compromised: acquisition time, restriction wavelength two-dimensional information, lack resolution larger views. elegantly new paper, these limitations partially overcome multiwavelength fixed fibroblast cells upscaling single-wavelength live red blood cells, allowing little than sixteen thousand single, view. Furthermore, extended study behaviour dense cell populations under flow conditions, movement topology no detectable photobleaching photodamage specimen. throughput lateral speed offers great advantages analysing heterogeneity phenotypes populations, making it promising tool diagnosis diseases affect shape. her review, Irina Kolotuev showcases versatility consists serial sections resin-embedded samples solid substrates such silicon wafers.6 beginning, section arrays were frequently analysed included EM high-end scanning microscopes offering unprecedented areas/volumes. review touches upon key steps workflow (namely sample preparation, embedding, trimming, sectioning, analysis), step compared those serial-sectioning TEM main SEM-based (serial block face focused ion beam SEM). Using examples very types originally processed TEM-based biology projects supported detailed highly informative illustrations supplementary movies, demonstrates (1) applicable most preparation protocols resins, (2) easy transfer multiple substrate, (3) offers, nondestructive method, reimaging series (of areas, magnification, microscopes), (4) provides choice thickness, (5) facilitates screening x, y z order randomly distributed scarce interest, (6) enables simultaneous BSE detectors z-resolution. makes strong claiming 'SEM can efficiently replace address many developmental questions'. rich source researchers interested workflows. Special Issue concludes in-depth Siân Culley, Alicia Cuber Caballero, Jemima J. Burden Virginie Uhlmann vast vague data sciences.7 aspects seem self-explanatory, Culley managed describe, define evaluate seemingly apparent notions. General concepts affecting measurements subsequent contrast generation modalities, photons electrons (considering binning, pixel size, under-/oversampling respect Nyquist–Shannon criterion, bit depth) segmentation processing and/or machine-learning tools explained illustrations. Based principle 'any extracted from good themselves', collaborators then distinguish quantitative bioimage information—intensity, morphology counts labels. Detailed definitions descriptions traits bioimaging given, again illustrations, methods how quantify them carefully evaluated, taking particularly care factors may influence thus quality any analysis. Information strategies deep-neural networks weaved chapters explaining potentials, challenges pitfalls quantification. put together impressive community comprehensive complex principles quantifying well data, completed assembling nonredundant feature vectors quality. It represents document scientific practice surely reference field.

Language: Английский

Introduction to women in microscopy: Volume 2 DOI
Michelle Peckham, Ulla Neumann, S J Culley

et al.

Journal of Microscopy, Journal Year: 2024, Volume and Issue: 295(1), P. 3 - 5

Published: June 11, 2024

As with the first volume of 'Women in Microscopy', we have assembled a wide-ranging collection papers focus on microscopy techniques and technologies encompassing five contributions topics as diverse surface passivation functionalisation for mass photometry applications, innovative time-modulated structured illumination single-molecule localisation microscopy, Mesolens imaging large fields view at low magnification yet high resolution, array tomography mainly but not exclusively 3D electron quantification life sciences microscopy. We would like to thank once more all authors two fantastic volumes Microscopy' their participation. The return rate after our initial call demonstrated interest this topic throughout community. second starts letter editor implementation an interdisciplinary bootcamp UC Berkeley intended promote supportive networking women alleviate gender-related inequities field microscopy.1 In early 1990s, when Ulla Neumann was undergraduate student, she asked practical course by male instructor German university if knew what photomultiplier was—at time didn't. And pointed out that kind information could evidently be found Brigitte, typical 'women's magazine'. Meagan Esbin Lena Blackman shows some progress has been made over last three decades. Still, given global only about 10% are late-stage careers, is excellent example building network focusing incoming students STEM. Through lectures, demos, nanorotations industry tours, opportunity connect role models different, or less public/private settings both academia industry, providing academic also career development support. future, specifically include trans gender nonconforming diversity initiatives. technical note Suliana Manley's group describes improvements glass coatings cover slips reduce nonspecific binding label-free, single-particle based interferometric scattering (iSCAT) microscopy.2 This technique relies interference between light scattered single particles reflected coverslip. When observing weakly scattering, proteins, detection signal above background noise caused impurities remains challenge. While various routinely used fluorescence studies, case photometry. described note, successfully PEG molecular weight (2 kDa) combination APTES (3-aminopropyltriethoxysilane), producing high-quality (smaller dynamic signal) evidenced equal numbers detected unbinding events different test proteins. Surface functionalisation, process which target molecules displayed surface, efficient using PEG-maleimide create covalent thioether bonds thiol groups comparison positively charging surfaces poly-L-lysine. contribution will useful applications other label-free techniques. One perpetual quest localise individual ever-increasing precision, bypassing diffraction limit light. work Sandrine Lévêque-Fort's temporal method changes improve precision.3 random subset fluorescent specific emitting excitation ('blinking') allows spatially analyse its point spread function (based models) calculate localisation. Acquisition thousands images, each recording molecules, reconstruct emitters entire analysed. paper presents approach shifted temporarily either xy xz resulting sinusoid, temporally modulated emission fluorophores depending position positional being carried phase signal. Key novel named ModLoc (Modulated Localisation) strategy acquire four intensity values necessary extract demodulation within same camera frame frequency, combined use digital micromirrors devices modulation galvanometric mirrors optical setup. With help combination, achieve twofold improvement precision conventional approaches enabling higher spatial excitation, adaption objective parameters possibility extend tracking studies. Gail McConnell's introduces us into world mesoscale standing wave imaging, combining axially (a sinusoidal pattern generated interfering beams) image formation through low-magnification, high-numerical aperture lens produce high-resolution images fluorescently labelled, three-dimensional structures 2D dataset.4 Thus far, so-called (NA 0.47, 4×), McConnell colleagues 2016,5 number modalities (confocal, sheet, colour bright-field, wide-field epifluorescence, FLIM). However, used, one compromised: acquisition time, restriction wavelength two-dimensional information, lack resolution larger views. elegantly new paper, these limitations partially overcome multiwavelength fixed fibroblast cells upscaling single-wavelength live red blood cells, allowing little than sixteen thousand single, view. Furthermore, extended study behaviour dense cell populations under flow conditions, movement topology no detectable photobleaching photodamage specimen. throughput lateral speed offers great advantages analysing heterogeneity phenotypes populations, making it promising tool diagnosis diseases affect shape. her review, Irina Kolotuev showcases versatility consists serial sections resin-embedded samples solid substrates such silicon wafers.6 beginning, section arrays were frequently analysed included EM high-end scanning microscopes offering unprecedented areas/volumes. review touches upon key steps workflow (namely sample preparation, embedding, trimming, sectioning, analysis), step compared those serial-sectioning TEM main SEM-based (serial block face focused ion beam SEM). Using examples very types originally processed TEM-based biology projects supported detailed highly informative illustrations supplementary movies, demonstrates (1) applicable most preparation protocols resins, (2) easy transfer multiple substrate, (3) offers, nondestructive method, reimaging series (of areas, magnification, microscopes), (4) provides choice thickness, (5) facilitates screening x, y z order randomly distributed scarce interest, (6) enables simultaneous BSE detectors z-resolution. makes strong claiming 'SEM can efficiently replace address many developmental questions'. rich source researchers interested workflows. Special Issue concludes in-depth Siân Culley, Alicia Cuber Caballero, Jemima J. Burden Virginie Uhlmann vast vague data sciences.7 aspects seem self-explanatory, Culley managed describe, define evaluate seemingly apparent notions. General concepts affecting measurements subsequent contrast generation modalities, photons electrons (considering binning, pixel size, under-/oversampling respect Nyquist–Shannon criterion, bit depth) segmentation processing and/or machine-learning tools explained illustrations. Based principle 'any extracted from good themselves', collaborators then distinguish quantitative bioimage information—intensity, morphology counts labels. Detailed definitions descriptions traits bioimaging given, again illustrations, methods how quantify them carefully evaluated, taking particularly care factors may influence thus quality any analysis. Information strategies deep-neural networks weaved chapters explaining potentials, challenges pitfalls quantification. put together impressive community comprehensive complex principles quantifying well data, completed assembling nonredundant feature vectors quality. It represents document scientific practice surely reference field.

Language: Английский

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