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Adobe these days launched a mid-cycle update to its eLearning Suite, a collection of equipment used to make tutorial content material for tutorial use and knowledgeable practicing. Updates to the suite, now in version 2.5, focal point on growing extra streamlined workflows, increasing output alternate options, and enhancing the software's usual usability.
The software is used by way of educational clothier and builders who create online or technologically delivered learning or practicing content. An example of e-gaining knowledge of content material that many company employees would locate regular is work safety and hazard practicing or sexual harassment practising, which may blend video clips, narrative storytelling, reading, and quizzes, all facilitated through a computer. E-studying is now used generally across better training, executive, businesses, and non-income agencies.
In one other new exchange, Adobe is letting shoppers license the eLearning Suite via paying for a month-to-month or every year subscription, because it did with its inventive Suite (version 5.5) a couple of weeks ago. The month-to-month plan even allows for purchasers to put their subscriptions on grasp and restart them as mandatory.
eLearning Suite 2.5 now contains Adobe Audition CS5.5, a robust substitute for Adobe Soundbooth that permits multitrack audio modifying on PCs and Macs. other updates to the application now let clients export audio for a complete Captivate 5.5 assignment with one click, significantly edit it in Audition CS5.5 and produce it back to Captivate while protecting slide affiliation.
On the visible side of issues, users can now create PDF-based mostly e-getting to know content the use of Acrobat X pro and post it to any number of leading discovering management methods.
New adjustments to Adobe Captivate 5.5 consist of being capable of put up content in MP4 structure, which could play on iOS and Android-based devices, create a video project using preset YouTube dimensions, draft quizzes with new templates, and more.
Adobe eLearning Suite 2.5 and Adobe Captivate 5.5 are available through Adobe licensed resellers and the Adobe store starting today. The estimated road cost for Adobe eLearning Suite 2.5 is $1,799 and $799 for Adobe Captivate 5.5. upgrade and training pricing for both items is additionally attainable.
Subscription pricing for eLearning Suite 2.5 starts at $89 per month for the one-year plan and $a hundred thirty five monthly for the month-to-month plan; subscription pricing for Adobe Captivate 5.5 is $39 per 30 days and $fifty nine monthly, respectively.
Adobe (http://www.adobe.com) has launched Adobe eLearning Suite 6, the enterprise’s integrated toolbox for developing knowledgeable-grade eLearning and HTML5-primarily based mLearning content.
The suite makes it possible for eLearning authorities, educators and trainers to create immersive, interactive eLearning classes finished with simulations, product demos, digital imaging, animations and audio production. Anchored via Adobe Captivate 6 and the brand new Adobe Presenter eight, Adobe eLearning Suite 6 comprises Adobe Flash knowledgeable CS6, Adobe Dreamweaver CS6, Adobe Photoshop CS6 extended and Adobe Acrobat X pro.
Adobe eLearning Suite 6 enhances productivity with round-tripping workflows between Adobe Captivate and Adobe Audition CS6, Adobe Flash knowledgeable CS6, Adobe Photoshop extended CS6 and Acrobat X seasoned, in keeping with Naresh Gupta, senior vice president, Print and Publishing, Adobe. as an instance, users can open Adobe Captivate information in Adobe Audition CS6 to alter speech pitch and alignment or Adobe Photoshop extended CS6 to retouch pictures and create 3D photos.
Prebuilt native extensions with Adobe Flash knowledgeable CS6 enable users to generate sprite sheets and greater. With Acrobat X seasoned users can embed Adobe Captivate videos into PDF documents and PDF portfolios, bringing text-primarily based getting to know materials to life, and make the accomplished undertaking accessible by way of Adobe Reader.
Adobe Presenter 8
the new Adobe Presenter eight empowers enterprise gurus, trainers and educators to create video shows correct from the computing device, devoid of the want for specialised machine or working towards, says Gupta. With Adobe Presenter 8, clients can streamline tasks and reduce expenses of manufacturing and editing videos with a single desktop solution, he provides. furthermore, they’re capable of concurrently capture slide shows, webcam video and audio and dynamically mix components with out a separate video editor.
Adobe Captivate 6
Adobe Captivate 6 enhancements improve cell entry and boost learner engagement while guaranteeing effortless publishing to leading SCORM- and AICC-compliant gaining knowledge of administration systems (LMSs) equivalent to Moodle, Blackboard, Plateau, Saba and SumTotal, in line with Gupta. It introduces the potential to submit content as each SWF and HTML5, enabling beginners to start a route on their desktop, pause and later resume on a cell gadget, together with iOS and Android primarily based capsules and smartphones.
clients can also create and edit HD-pleasant demos in the new “capture-as-a-video” workflow, insert videos in an image-in-picture format and publish tasks to YouTube — all within the identical consumer interface. A library of out-of-the-box belongings, more advantageous Microsoft PowerPoint integration and better quizzing capabilities complete the desirable new features in Adobe Captivate 6.
Adobe eLearning Suite 6 is automatically obtainable via Adobe authorized resellers and the Adobe store for an estimated street price of US$1,799. users of eLearning Suite 2 and 2.5 can upgrade at a reduced cost of $599. improve pricing from Adobe Captivate 5.5, 5 or four to Adobe eLearning Suite 6 is $1,199. certified training users can purchase eLearning Suite 6 for US$599. For a complimentary trial, visit http://www.adobe.com/products/elearningsuite/trial.html .
New HD Video Workflow, other Key Enhancements assist Trainers right now carry expert-Grade eLearning content material
London. - June, 15 2012 - Adobe techniques incorporated (Nasdaq:ADBE) nowadays announced the immediate availability of Adobe® Captivate® 6, a major improve to its business-leading eLearning authoring software for swiftly growing a wide array of interactive eLearning and HTML5-based mostly mobile discovering content material. Designed with cutting-edge cell beginners in mind, Captivate 6 allows for eLearning developers, corporate trainers, educators and different enterprise clients to support bring dynamic, SCORM- and AICC-compliant course content material this is obtainable anytime, anyplace.
"nowadays, freshmen are expecting attractive eLearning alternate options on their iOS and Android gadgets - static displays gotten smaller to fit mobile displays don't seem to be ample," pointed out Naresh Gupta, senior vice chairman, Print and Publishing, Adobe. "Captivate 6 offers discipline matter consultants and content creators the ability to bring eLearning content to cellular devices it really is as potent and interactive because the content brought to pcs."
Captivate 6 enhancements enhance cell access and enhance learner's engagement - properly points consist of:
With Captivate 6, trainers and educators can individualise eLearning modules by recording voiceovers and other sounds that immediately play returned when a learner clicks on a designated object. route designers can additionally be sure that eLearning content material continues a constant seem and believe the usage of customisable, professionally designed topics. stronger LMS integration helps eLearning developers quite simply submit content to leading gaining knowledge of administration systems, including Moodle, Blackboard, Plateau, Saba and SumTotal.
Dustin Tauer, vice president, practicing and construction, Easel options
Eric Fields, senior eLearning consultant, gaining knowledge of and construction, Coventry health Care workers' Comp functions
Damien Bruyndonckx, multimedia assistant, IHECS, Haute Ecole Gallilee
Pricing and Availability
Adobe Captivate 6 is immediately available through Adobe accredited resellers and the Adobe shop for an estimated road rate of £714 (ex VAT). Captivate 5.5 and Captivate four clients can upgrade at a discounted price of £286 (ex VAT) and £429 (ex VAT), respectively. certified schooling clients should buy Captivate 6 for £238 (ex VAT). For a free trial, discuss with www.adobe.com/go/trycaptivate.
About Adobe systems integrated
Adobe is changing the realm through digital experiences. For more assistance, talk over with www.adobe.com/uk.
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Adobe (http://www.adobe.com) has released Adobe eLearning Suite 6, the company’s integrated toolbox for creating professional-grade eLearning and HTML5-based mLearning content.
The suite enables eLearning professionals, educators and trainers to create immersive, interactive eLearning courses complete with simulations, product demos, digital imaging, animations and audio production. Anchored by Adobe Captivate 6 and the new Adobe Presenter 8, Adobe eLearning Suite 6 includes Adobe Flash Professional CS6, Adobe Dreamweaver CS6, Adobe Photoshop CS6 Extended and Adobe Acrobat X Pro.
Adobe eLearning Suite 6 enhances productivity with round-tripping workflows between Adobe Captivate and Adobe Audition CS6, Adobe Flash Professional CS6, Adobe Photoshop Extended CS6 and Acrobat X Pro, according to Naresh Gupta, senior vice president, Print and Publishing, Adobe. For example, users can open Adobe Captivate files in Adobe Audition CS6 to adjust speech pitch and alignment or Adobe Photoshop Extended CS6 to retouch images and create 3D graphics.
Prebuilt native extensions with Adobe Flash Professional CS6 enable users to generate sprite sheets and more. With Acrobat X Pro users can embed Adobe Captivate movies into PDF documents and PDF portfolios, bringing text-based learning materials to life, and make the finished project accessible via Adobe Reader.
Adobe Presenter 8
The new Adobe Presenter 8 empowers business professionals, trainers and educators to create video presentations right from the desktop, without the need for specialized equipment or training, says Gupta. With Adobe Presenter 8, users can streamline projects and lower costs of producing and editing videos with a single desktop solution, he adds. In addition, they’re able to simultaneously capture slide presentations, webcam video and audio and dynamically mix components without a separate video editor.
Adobe Captivate 6
Adobe Captivate 6 enhancements improve mobile access and boost learner engagement while ensuring effortless publishing to leading SCORM- and AICC-compliant Learning Management Systems (LMSs) such as Moodle, Blackboard, Plateau, Saba and SumTotal, according to Gupta. It introduces the ability to publish content as both SWF and HTML5, enabling learners to begin a course on their desktop, pause and later resume on a mobile device, including iOS and Android based tablets and smartphones.
Users can also create and edit HD-quality demos within the new “capture-as-a-video” workflow, insert videos in a picture-in-picture format and publish projects to YouTube — all within the same user interface. A library of out-of-the-box assets, improved Microsoft PowerPoint integration and enhanced quizzing capabilities complete the top new features in Adobe Captivate 6.
Adobe eLearning Suite 6 is immediately available through Adobe authorized resellers and the Adobe Store for an estimated street price of US$1,799. Users of eLearning Suite 2 and 2.5 can upgrade at a discounted price of $599. Upgrade pricing from Adobe Captivate 5.5, 5 or 4 to Adobe eLearning Suite 6 is $1,199. Qualified education users can purchase eLearning Suite 6 for US$599. For a complimentary trial, visit http://www.adobe.com/products/elearningsuite/trial.html .
The AIPLA has now offered its legislative proposal for rewriting 35 U.S.C. § 101 that is quite close to that offered by the IPO:
(a) Eligible Subject Matter.—Whoever invents or discovers any new and useful process, machine, manufacture, composition of matter, or any useful improvement thereof, may obtainshall be entitled to a patent therefor, subject only to the conditions and requirements ofset forth in this title.
(b) Sole Exceptions to Subject Matter Eligibility.—A claimed invention is ineligible under subsection (a) only if the claimed invention as a whole exists in nature independent of and prior to any human activity, or can be performed solely in the human mind.
(c) Sole Eligibility Standard.—The eligibility of a claimed invention under subsections (a) and (b) shall be determined without regard to the requirements or conditions of sections 102, 103, and 112 of this title, the manner in which the claimed invention was made or discovered, or whether the claimed invention includes an inventive concept.
AIPLA statement. The AIPLA proposal is strikingly similar to that of the IPO’s (although not acknowledged by the AIPLA statement).
[DOCX File of Table: Comparing101Proposals]
The research site near the town of Rifle, northwestern Colorado (USA), has been described previously42. Briefly, the site is located on a 9 ha floodplain in northwestern Colorado that is underlain by an aquifer comprised of 6–7 m of unconsolidated sands, silts, clays and gravels deposited by the Colorado River. Amendment of acetate to the aquifer occurred through five boreholes oriented orthogonal to groundwater flow direction and spaced at 1.5-m intervals. Cross-well mixing was used to disperse the injectate across the width of the injection zone.
Groundwater samples were taken prior to (GWA1) and following acetate amendment (GWB1). Acetate-amended groundwater was injected upgradient 3.5 and 5.5 m below the surface to achieve aquifer concentrations of 15 mM (acetate; Sigma-Aldrich, Saint Louis, MO, USA) and 2 mM (bromide; Sigma-Aldrich). Prior to acetate amendment 140 l, and on 03 September 2011 and 05 September 2011, 7 days (GWB1) and 9 days after the start of acetate amendment, 100 l of groundwater were pumped and filtered sequentially through a 1.2-μm pore size pre-filter (293-mm diameter Supor-1200 hydrophilic polyethersulfone membrane disc filter; Pall Corporation, Ann Arbor, MI, USA), with biomass retained on a 0.2-μm pore size (293-mm diameter Supor-200 hydrophilic polyethersulfone membrane disc; Pall Corporation) and a 0.1-μm pore size sample filter (142-mm diameter Supor-100 hydrophilic polyethersulfone membrane disk filter; Pall Corporation). Filters were immediately frozen in an ethanol–dry ice mix, stored at −80 °C and shipped overnight to the University of California, Berkeley, for DNA extraction. For cryo-TEM, 500 ml of 0.2-μm filtrate was concentrated with Vivaspins (cutoff 30 kDa; GE Healthcare, Pittsburgh, PA, USA) to ~500 μl and cryo-plunged immediately (see below). For molecular, metagenomic and cryo-TEM correlation analyses, the same groundwater sample (GWB1) was used.DNA extractions
Approximately 1 g of each filter was used for DNA extraction using the PowerMax Soil DNA Isolation kit (Mo Bio Laboratories Inc., Carlsbad, CA, USA, Cat# 12988). Manufacturer’s protocol was followed, with the exception of adding a freeze/thaw step and vortexing bead tubes for 3.5 min after addition of the SDS reagent, followed by 30 min at 65 °C with intermittent shaking. DNA in the 5-ml eluted volume was concentrated by sodium acetate/ethanol precipitation with glycogen followed by resuspension in provided elution buffer.Preparation of clone libraries and sequencing
Full-length, bacterial 16S rRNA sequences were amplified by utilizing a gradient PCR using general bacterial primers 27F (5′-AGAGTTTGATCMTGGCTCAG-3′) and 1492 R (5′-GGTTACCTTGTTACGACTT-3′)43. For PCR, the thermocycler reaction conditions were as follows: initial denaturation at 94 °C for 1 min, 25 cycles of denaturation at 94 °C for 30 s, annealing across an eight-step gradient from 48–59 °C for 30 s, extension at 72 °C for 1 min and a final extension at 72 °C for 7 min. Correct amplicon size was verified with gel electrophoresis and the PCR product was cleaned up using the UltraClean PCR Clean-up Kit (Mo Bio Laboratories Inc., CA Cat# 12500). Clone libraries were generated using a TOPO TA cloning kit and electrocompetent cells (Life Technologies Corp., Grand Island, NY, USA). One hundred transformants from the 0.1- and 0.2-μm clone libraries were verified by colony PCR using the M13 forward (5′-GTAAAACGACGGCCAGT-3′) and reverse (5′-CAGGAAACAGCTATGAC-3′) primers and gel electrophoresis. The colony PCR thermocycler amplification conditions were as follows: E. coli cell lysis and initial denaturation at 95 °C for 10 min, 25 cycles of denaturation at 95 °C for 30 s, annealing at 53 °C for 30 s and extension at 72 °C for 1.5 min and a final extension at 72 °C for 7 min. Successful transformants were Sanger sequenced using the M13 forward and reverse primers (only for the 0.1-μm filter). Sequences were primer and vector screened using cross_match ( http://www.phrap.org) and NCBI VecScreen ( http://www.ncbi.nlm.nih.gov/VecScreen/VecScreen.html), quality scored using Phred ( http://www.phrap.org) and assembled into contigs using Phrap ( http://www.phrap.org). Sequences were trimmed to retain only bases Phred ≥q20 and high-quality contigs were tested for chimeras using USEARCH 64 ( http://www.drive5.com). Sequences were identified utilizing BLAST44 against the Arb-Silva Database ( http://www.arb-silva.de).16S rRNA gene phylogenetic analysis
16S rRNA gene sequences from cells retained on the 0.2 μm filter (50 clones, resulting in 21 operational taxonomic units (OTUs) after chimera checking and clustering as described previously) and 0.1-μm filter (108 clones, resulting in 24 OTUs) were obtained by sequencing of the clone libraries. The individual clone sequences were clustered at 97% using UCLUST (part of USEARCH 64). We also used EMIRGE20 to reconstruct 16S rRNA gene sequences after trimming the Illumina reads using sickle to remove low-quality bases ( https://github.com/najoshi/sickle). For EMIRGE, paired-end reads, where both reads were at least 60 nucleotides in length after trimming, were used as inputs. For each sample, EMIRGE was run for 100 iterations. Reconstructed sequences for all sampled taxa were combined with database sequences representing the most closely related taxa for subsequent analysis. EMIRGE reconstructions generated 26 and 36 OTUs for the 0.2- and 0.1-μm filters, respectively. EMIRGE, clone library and Arb-Silva database WWE3-OP11-OD1 16S rRNA gene sequences were aligned with MUSCLE45 using default parameters. The alignment was used to generate a maximum likelihood tree with RAxML46 using the GTRCAT model of nucleotide substitution and 200 bootstrapped replicates and E. coli as an outgroup. The tree was edited using iTOL47. Poorly aligned or lower-quality sequences from the Arb-Silva database were removed prior to further analysis. The environments from which each sequence was obtained were pulled from the Arb-Silva database using the Arb software package.Metagenomics methods
A total of 9,781,022,700 bp of Illumina data (150 bp paired reads) was generated for GWA1 and 369,257,200 bp was generated for GWB1at the Joint Genome Institute, Walnut Creek, CA. The same GWB1 sample (0.1-μm filter fraction) was used for cryo-TEM characterization. Sequence data sets were assembled (after trimming to remove low-quality bases) using idba_ud48 using the default settings. Open-reading frames were predicted using Meta-Prodigal49 and assigned a preliminary annotation using USEARCH44 against the Uniref90 database ( http://www.uniprot.org/). Community composition was profiled primarily using single-copy ribosomal protein S3 genes carried on scaffolds >5 kb in length (detection limit ~0.01%). Organism abundance levels were determined based on sequence coverage. Detailed genome reconstructions for the organisms in these samples will be reported separately.
Because sequences from the most abundant populations (high sequence coverage) often assemble poorly, the analysis also used two data subsets per sample (1/10th and 1/50th of the data for the GWB1 sample and 1/9th and 1/27th of the data for the GWA1 sample). Community composition analysis used results reconciled from these subassemblies. Genomic data from the subassemblies were binned to specific populations based on GC content, coverage and phylogenetic profile. Each genome was either near-complete or well sampled in one or multiple data sets. Phylogenetic profiling-based binning was helpful because many organisms on the filtrates were relatively similar to organisms that are represented in our in-house candidate phyla genomic data set (WWE3, OP11, OD1 and archaea: reported in refs 14, 16, and data to be published elsewhere). Abundances are reported as coverage and/or DNA representation. Coverage was determined based on read mapping statistics. DNA representation used coverage statistics, approximate genome size and total data size (as above).Cryo-TEM specimen preparation in the field
For cryo-TEM and synchrotron infrared (SIR) spectromicroscopy (see below), 200 mesh lacey carbon-coated formvar Cu-grids (Ted Pella Inc., Redding, CA, USA) were used. For correlative FISH and TEM, a lacey or a continuous formvar support film was laid on TEM nickel finder grids (Maxtaform Finder Grid Style H7, 63-μm pitch 400 mesh) and grids were carbon coated. All TEM grids were treated by glow discharge to improve sample deposition onto the grids. Ten and 250 nm colloidal gold particles (BBInternational, Cardiff, UK) were put on TEM grids for cryo-TEM and SIR spectroscopy, and for correlative FISH and TEM, respectively, and allowed to dry prior to sample addition. Aliquots of 5 μl 0.2-μm-filtered groundwater sample were deposited onto the grids, manually blotted with filter paper and plunged into liquid propane at liquid nitrogen temperature using a portable cryo-plunge device on site17. Grids were stored in liquid nitrogen until further analysis.Clone fluorescence in situ hybridization
Subcloning for construction of the positive controls, E. coli cells each carrying the 16S rRNA gene sequence of one of the three bacterial types (WWE3, OP11 and OD1) was performed using the Novagen AccepTor Vector Kit (EMD Millipore, Merck KGaA, Darmstadt, Germany). Subclones with OP11-WWE3-OD1 16S rRNA gene sequences present were identified by sequencing using pETBlueT7UP forward (5′-TCATAACGTCCCGCGAAA-3′) and pETBlueDown reverse (5′-GTTAAATTGCTAACGCAGTCA-3′) primers and BLAST44 against the Arb-Silva Database. Plasmids containing WWE3-OP11-OD116S rRNA sequences were isolated from subclones used to transform into the NovaBlue (DE3) strain for the subsequent Clone-FISH steps (EMD Millipore, Merck KGaA).
Clone-FISH E. coli strains transformed with WWE3-OP11-OD1 sequences were fixed for FISH by centrifuging at 15,000 r.p.m. for 2 min at 4 °C, resuspending in 1 ml PBS (pH 7), centrifuging again and resuspending in 250 μl PBS and 750 μl 4% paraformaldehyde. Cells were allowed to fix for 3 h at 4 °C before centrifuging at 15,000 r.p.m. for 2 min at 4 °C, resuspending in a 1:1 mixture of ethanol and PBS. FISH runs were performed at a range of formamide concentrations between 20 and 50% to establish the optimum concentration that allowed proper hybridization but reduced apparent nonspecific binding.CARD-FISH
For correlative cryo-TEM and CARD-FISH two approaches were performed. For the first approach, frozen samples on Ni-Finder TEM grids were imaged and then the CARD-FISH protocol was applied50. For the second approach, frozen samples on Ni-Finder TEM grids were freeze-dried and embedded in low-gelling point agarose (0.1% final concentration), dried at room temperature, then fixed in paraformaldehyde solution (2% final concentration), washed in sterile Milli-Q water, dehydrated in 50, 80, 90 and in 100% ethanol and air dried. Three different oligonucleotide probes (Supplementary Table 8), targeting rRNA genes, were applied to cells on TEM grids. Hybridization was performed following a method previously described in ref. 50, with a formamide concentration of 50%, incubation at 46 °C for 3 h and washing at 48 °C for 10 min. The subsequent amplification was performed at 46 °C for 10 min. Samples were counterstained with 4',6-diamidino-2-phenylindole DNA stain (1 μg ml−1 final concentration).
Confocal laser scanning microscopy was performed on a Carl Zeiss Inc. LSM 710 Zen 2010, Release Version 6.0 software (Carl Zeiss MicroImaging Inc., Thornwood, NY, USA), equipped with argon (458 nm, 488 nm and 514 nm) and He–Ne (594 nm, 543 nm and 633 nm) lasers and a diode 45–30 (405 nm). The diode (405 nm) was used for 4',6-diamidino-2-phenylindole signals (BP filter 410–585). Positively labelled cells (fluorochrome Alexa Fluor 546) were detected using the He–Ne 543 nm laser line (BP filter 548–680). A Plan-Apochromat × 100/1.4 oil differential interference contrast (DIC) (Zeiss) lens was used.2D and 3D cryo-TEM
Cryo-TEM images were acquired on a JEOL–3100-FFC electron microscope (JEOL Ltd, Akishima, Tokyo, Japan) equipped with a field emission gun electron source operating at 300 kV, an Omega energy filter (JEOL), cryo-transfer stage and a Gatan 795 4 × 4 K charge-coupled device camera (Gatan Inc., Pleasanton, CA, USA) mounted at the exit of an electron decelerator held at a voltage of 200–250 kV51. The stage was cooled with liquid nitrogen to 80 K during acquisition of all data sets.
Over 100 2D images were recorded at different magnifications giving a pixel size of 0.375, 0.28 or 0.22 nm at the specimen. Underfocus values ranged between 3.6 μm±0.25 μm and 12 μm±0.5 μm, and energy filter widths were typically around 30 eV. The survey of the grids and the selection of suitable targets were done in low-dose defocused diffraction mode to minimize radiation damage.
Thirteen tomographic tilt series were acquired under low-dose conditions, typically over an angular range between +65° and −65°, ±5° with increments of 2°. Between 61 and 66 images were recorded for each tilt series, acquired semi-automatically with the program Serial-EM ( http://bio3d.colorado.edu/)52 adapted to JEOL microscopes. For tilt series data sets, all images show a pixel size of 0.56 or 0.746 nm at the specimen. Underfocus values ranged between 3.6 μm±0.25 μm and 9 μm±0.5 μm, and energy filter widths were ~30 eV. The average dose used per complete tilt series was ~113 e− Å−2. All tomographic reconstructions were obtained with the program Imod ( http://bio3d.colorado.edu/)52. The software ImageJ 1.38 × (NIH, http://rsb.info.nih.gov/ij/)53 was used for analysis of the 2D image projections. All movies were created with the open-source package ffmpeg ( http://www.ffmpeg.org/). Adobe Photoshop CS5.1 was used to adjust contrast in the images and to insert calibrated scale bars into images.SIR spectromicroscopy
Cryo-TEM grids were placed onto the BaF2 infrared windows (International Crystal Laboratories, NJ, USA) under liquid nitrogen. They were then allowed to air dry at ambient temperature on the BaF2 windows.
SIR spectromicroscopy was performed at the infrared beamline 1.4.3 (Advanced Light Source, http://infrared.als.lbl.gov/) on a Nic-Plan infrared microscope ( × 32 objective, numerical aperture=0.65; released software OMNIC 7.0) equipped with a Nicolet Magna 760 infrared spectrometer (Thermo Scientific Inc., MA, USA) at the mid-infrared frequency range (2.5–15.5 μm wavelength, or 4,000–650 cm–1 wavenumber). The infrared signals (in absorbance) from the energy exchange between the infrared photons and biomolecules were sampled by dividing the TEM grid in 2-μm pixels, raster scanned and processed following a method previously described elsewhere22,54. Cells were detected using the absorption bands of protein amide I and of lipids methyl (−CH3) and methylene (−CH2−) groups. Analysis made use of a database of known bacterial and archaeal standards.
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