{"id":11517,"date":"2024-09-26T10:15:13","date_gmt":"2024-09-26T10:15:13","guid":{"rendered":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/?page_id=11517"},"modified":"2024-10-18T17:52:46","modified_gmt":"2024-10-18T17:52:46","slug":"automated-saxs","status":"publish","type":"page","link":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/home\/sample-environments\/automated-saxs\/","title":{"rendered":"Automated SAXS"},"content":{"rendered":"\n<div class=\"embl-grid\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p class=\"vf-text--body vf-text-body--1\"><strong>Automated BioSAXS at P12 <\/strong><\/p>\n\n\n\n<p>allows users to perform experiments at a <strong>high turnover rate<\/strong>, including data collection and downstream processing. <\/p>\n\n\n\n<p>Experiments are possible <strong>on-site or by mail-in access<\/strong>, where the beamline staff conducts them. <\/p>\n\n\n\n<p>Three modules can be operated simultaneously:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Static SAXS<\/li>\n\n\n\n<li>SEC coupled SAXS<\/li>\n\n\n\n<li>AF4-SAXS<\/li>\n<\/ul>\n\n\n\n<p>Samples are measured in the <strong>vacuum Sample Exposure Uni<\/strong>t, which houses a <strong>quartz capillary with a 1 or 2 mm diameter<\/strong><\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<article class=\"vf-card vf-card--brand vf-card--bordered vf-u-margin__bottom--800\" default>\n  <div class=\"vf-card__content | vf-stack vf-stack--400\">\n      <h3 class=\"vf-card__heading\">\n      Batch SAXS (static)    <\/h3>\n            <p class=\"vf-card__subheading\">automated auto sampler<\/p>\n          <\/div>\n<\/article>\n\n\n\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"770\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-1024x770.png\" alt=\"\" class=\"wp-image-11519\" srcset=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-1024x770.png 1024w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-300x225.png 300w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-768x577.png 768w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS.png 1187w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p><\/p>\n\n\n\n<p><strong>Specifications:<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Minimum Volume:<\/strong> 10 \u00b5l<\/li>\n\n\n\n<li><strong>Recommended Volume:<\/strong> 50 \u00b5L (100 \u00b5l for complete concentration series)<\/li>\n\n\n\n<li><strong>Measurement cycle:<\/strong> 90 seconds<\/li>\n\n\n\n<li><strong>Data analysis:<\/strong> &lt; 3min after measurement<\/li>\n\n\n\n<li><strong>Temperature Range:<\/strong>\n<ul class=\"wp-block-list\">\n<li><strong>Cell:<\/strong> 7\u00b0C to 60\u00b0C (up to 80\u00b0C for short periods)<\/li>\n\n\n\n<li><strong>Tray:<\/strong> 7\u00b0C to 40\u00b0C<\/li>\n<\/ul>\n<\/li>\n\n\n\n<li><strong>Compatible Sample Formats (all available on-site):<\/strong>\n<ul class=\"wp-block-list\">\n<li>Eppendorf strip (4 x 8 slots)<\/li>\n\n\n\n<li>1.5 mL lidless Eppendorf tubes (12 slots)<\/li>\n\n\n\n<li>2 x 96-well plates<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n\n\n\n<p><\/p>\n\n\n\n<p><a href=\"#batch-saxs-2\" data-type=\"internal\" data-id=\"#batch-saxs-2\">LEARN MORE<\/a><\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<article class=\"vf-card vf-card--brand vf-card--bordered vf-u-margin__bottom--800\" default>\n  <div class=\"vf-card__content | vf-stack vf-stack--400\">\n      <h3 class=\"vf-card__heading\">\n      SEC-(MALS)-SAXS    <\/h3>\n            <p class=\"vf-card__subheading\">size exclusion coupled SAXS<\/p>\n          <\/div>\n<\/article>\n\n\n\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"778\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-2-1024x778.png\" alt=\"\" class=\"wp-image-11559\" srcset=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-2-1024x778.png 1024w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-2-300x228.png 300w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-2-768x584.png 768w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-2.png 1192w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p><\/p>\n\n\n\n<p><strong>Specifications:<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Injection Volume:<\/strong> 20 &#8211; 100 \u00b5l<\/li>\n\n\n\n<li><strong>Concentration:<\/strong> 5-10 mg\/ml<\/li>\n\n\n\n<li><strong>Measurement length:<\/strong> 50 min for 24 ml columns; 15min for 3 ml columns<\/li>\n\n\n\n<li><strong>Data analysis:<\/strong> frames available for Chromixs analysis ~ 5 min after measurement<\/li>\n\n\n\n<li><strong>Temperature Range:<\/strong>\n<ul class=\"wp-block-list\">\n<li><strong>Autosampler:<\/strong> 4\u00b0C to 40\u00b0C<\/li>\n\n\n\n<li><strong>Column:<\/strong> Room temperature<\/li>\n<\/ul>\n<\/li>\n\n\n\n<li><strong>Important information:<\/strong>\n<ul class=\"wp-block-list\">\n<li>Sufficient mobile phase (500 ml) required<\/li>\n\n\n\n<li><a href=\"#available-columns\" data-type=\"internal\" data-id=\"#available-columns\">Columns available<\/a> at the beamline (upon request)<\/li>\n\n\n\n<li>Addition of MALS detectors (upon request)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n\n\n\n<p><\/p>\n\n\n\n<p><a href=\"#sec-saxs\" data-type=\"internal\" data-id=\"#sec-saxs\">LEARN MORE<\/a><\/p>\n\n\n\n<p><\/p>\n\n\n\n<p><\/p>\n\n\n\n<p><\/p>\n\n\n\n<p><\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<article class=\"vf-card vf-card--brand vf-card--bordered vf-u-margin__bottom--800\" default>\n  <div class=\"vf-card__content | vf-stack vf-stack--400\">\n      <h3 class=\"vf-card__heading\">\n      AF4-SAXS    <\/h3>\n            <p class=\"vf-card__subheading\">AF4 prior to SAXS<\/p>\n          <\/div>\n<\/article>\n\n\n\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"780\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-1-1024x780.png\" alt=\"\" class=\"wp-image-11557\" srcset=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-1-1024x780.png 1024w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-1-300x228.png 300w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-1-768x585.png 768w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-1.png 1187w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure>\n\n\n\n<p><\/p>\n\n\n\n<p><strong>Specifications:<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Injection Volume: <\/strong>20 &#8211; 1000 \u00b5l<\/li>\n\n\n\n<li><strong>Concentration:<\/strong> dependent on sample type<\/li>\n\n\n\n<li><strong>Measurement length:<\/strong> dependent on established method, typically between 35- 80 min<\/li>\n\n\n\n<li><strong>Data analysis:<\/strong> frames available for Chromixs analysis ~ 5min after measurement <\/li>\n\n\n\n<li><strong>Important information:<\/strong>\n<ul class=\"wp-block-list\">\n<li>Established separation method required (to be prearranged with beamline staff)<\/li>\n\n\n\n<li>standard channel and preparative channel available<\/li>\n\n\n\n<li>Sufficient mobile phase (0.5 &#8211; 2 l) required<\/li>\n\n\n\n<li>Financed through: BMBF (F\u00f6rderkennzeichen: 05K22UM3)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n\n\n\n<p><a href=\"#af4-saxs\" data-type=\"internal\" data-id=\"#af4-saxs\">LEARN MORE<\/a><\/p>\n\n\n\n<p><\/p>\n\n\n\n<p><\/p>\n\n\n\n<p><\/p>\n\n\n\n<p><\/p>\n\n\n\n<p><\/p>\n\n<\/div>\n<\/div>\n<\/div>\n\n\n<hr class=\"vf-divider\">\n\n\n<div class=\"embl-grid\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p id=\"batch-saxs-2\"><\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<article class=\"vf-card vf-card--brand vf-card--bordered vf-u-margin__bottom--800\" default>\n  <div class=\"vf-card__content | vf-stack vf-stack--400\">\n      <h3 class=\"vf-card__heading\">\n      BATCH SAXS    <\/h3>\n              <\/div>\n<\/article>\n\n\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"embl-grid\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"770\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-2-1024x770.png\" alt=\"\" class=\"wp-image-11605\" srcset=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-2-1024x770.png 1024w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-2-300x225.png 300w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-2-768x577.png 768w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1_b_BATCH_SAXS-2.png 1187w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><figcaption class=\"vf-figure__caption\">Left: Automated sample changer for efficient small-volume loading.<br>Right top: Liquid held in the capillary. Right bottom: Workflow from sample loading, measurement, to results in under 3 minutes.<\/figcaption><\/figure>\n\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"embl-grid\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p>Batch measurements are conducted using the sample changer, a liquid-handling robot designed for SAXS data collection on protein solutions at equilibrium (<strong>EMBL-ARINAX<\/strong>). <\/p>\n\n\n\n<p>The device can load as little as 5 \u00b5L of solution into a vacuum flow-through capillary, exposing the sample to the X-ray beam. <\/p>\n\n\n\n<p>The sample<strong> continuously flows<\/strong> through the capillary to minimize radiation damage during exposure (<strong>20 ul <\/strong>loading volume required). <\/p>\n\n\n\n<p>Between measurements, the system automatically cleans and dries the sample cell. A <strong>full cycle\u2014including sample loading, exposure, cell cleaning, and drying<\/strong>\u2014takes approximately <strong>90 seconds.&nbsp;<\/strong><\/p>\n\n\n\n<p>The <strong>temperature <\/strong>of the sample cell and tray, where samples are stored before measurement, can be <strong>individually controlled.<\/strong> <\/p>\n\n\n\n<p>The sample tray holds blocks for PCR strips (8 x 200 \u00b5l), 1.5ml tubes and 96-well plates. These <strong>consumables are available<\/strong> at the beamline.<\/p>\n\n\n\n<p>A typical batch experiment involves <strong>measuring the sample at different concentrations<\/strong>, along with the <strong>corresponding buffer<\/strong>. Buffers are typically measured between each sample concentration to ensure accuracy.<\/p>\n\n\n\n<p><strong>Exposure times <\/strong>range from <strong>1 to 5 seconds<\/strong>, depending on the sample&#8217;s radiation sensitivity. During this time,<strong> 20 to 50 frames<\/strong> are collected <strong>to identify any radiation damage<\/strong> so that compromised data can be discarded as needed.&nbsp;<\/p>\n\n<\/div>\n<\/div>\n<\/div>\n\n\n<hr class=\"vf-divider\">\n\n\n<div class=\"embl-grid\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p id=\"sec-saxs\"><\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<article class=\"vf-card vf-card--brand vf-card--bordered vf-u-margin__bottom--800\" default>\n  <div class=\"vf-card__content | vf-stack vf-stack--400\">\n      <h3 class=\"vf-card__heading\">\n      SEC-SAXS  and SEC-MALS\/SAXS    <\/h3>\n              <\/div>\n<\/article>\n\n\n\n\n<figure class=\"vf-figure wp-block-image size-large is-resized\"><img loading=\"lazy\" decoding=\"async\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-3-1024x778.png\" alt=\"\" class=\"wp-image-11653\" width=\"512\" height=\"389\" srcset=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-3-1024x778.png 1024w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-3-300x228.png 300w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-3-768x584.png 768w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1c_SEC_SAXS-3.png 1192w\" sizes=\"auto, (max-width: 512px) 100vw, 512px\" \/><figcaption class=\"vf-figure__caption\">Left: Setup of HPLC system at P12 hutch with adjacent detectors. Right top: Scheme illustrating the separation process. Right bottom: Elution profile highlighting larger particles eluting before smaller particles.<\/figcaption><\/figure>\n\n\n\n<p>Size exclusion chromatography, in line with SAXS, <strong>SEC-SAXS<\/strong>, features an<strong> Agilent 1260 Infinity II Bio-Inert HPLC system<\/strong> with a <strong>temperature-controlled autosampler <\/strong>and a <strong>UV detector<\/strong> is a variable wavelength detector (VWD) ranging from 190-600 nm.<\/p>\n\n\n\n<p>We<strong> provide Cytiva Superdex\u2122 or Superose\u2122 increase prepacked SEC columns.<\/strong> However, Users are<strong> encouraged to bring their column<\/strong>s, especially with specialized solvents, samples, or other columns containing different chromatography resins. <\/p>\n\n\n\n<p>Duration of SEC-SAXS experiment is dependent on column separation. It ranges from<strong> 15 min (for 3ml column<\/strong>) to <strong>50 min (24 ml column)<\/strong>. During the whole elution process, SAXS frames are collected, where the <strong>exposure time<\/strong> ranges from <strong>0.5 to 1 second<\/strong>. <\/p>\n\n\n\n<p>The <strong>recommended injection<\/strong> can be<strong> between 20-100 \u00b5l <\/strong>depending on the sample concentration and selected column. As the sample is diluted during the separation process, higher sample concentrations are required, around <strong>5-10 mg\/ml<\/strong>. All measurements are possible at room temperature only. <\/p>\n\n\n\n<p>In addition, we offer<strong> additional molecular weight validation from multi-angle laser light scattering (MALS) <\/strong>coupled to hydrodynamic radius measurements from dynamic light scattering (DLS) with the <strong>Wyatt miniDAWN TREOS\/QELS<\/strong> instrument and a <strong>Optilab\u2122 T-rEX <\/strong>refractive index (RI) instrument. When using the 24ml, this is done by splitting the eluent equally between detector array and SAXS capillary.<\/p>\n\n\n\n<p>More information here:<\/p>\n\n\n\n<blockquote class=\"vf-blockquote\">\n<h3 class=\"wp-block-heading\"><a href=\"https:\/\/www.doi.org\/10.3390\/cryst10110975\">Adding size exclusion chromatography (SEC) and light scattering (LS) devices to obtain high-quality small angle X-ray scattering (SAXS) data.<\/a><\/h3>\n\n\n\n<p>Graewert MA, Da Vela S, Gr\u00e4wert TW, Molodenskiy DS, Blanchet CE, Svergun DI, Jeffries CM<\/p>\n\n\n\n<p>Crystals,&nbsp;2020<\/p>\n\n\n\n<p>doi:10.3390\/cryst10110975.<\/p>\n<\/blockquote>\n\n<\/div>\n<\/div>\n<\/div>\n\n\n<hr class=\"vf-divider\">\n\n\n<div class=\"embl-grid\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p id=\"available-columns\"><\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p><strong>List of available columns provided:<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Superdex 200 Increase 10\/300: general purpose separation across a wide MW range (10-600 kDa).&nbsp;<\/li>\n\n\n\n<li>Superdex 200 Increase 5\/150<\/li>\n\n\n\n<li>Superdex 75 Increase 10\/300 (for small macromolecules less than 70 kDa).<\/li>\n\n\n\n<li>Superdex 75 Increase 5\/150<\/li>\n\n\n\n<li>Superose 6 Increase 10\/300 (for very-large macromolecules and complexes up to 5 MDa).<\/li>\n\n\n\n<li>Superose 6 Increase 5\/150 (for very-large macromolecules and complexes up to 5 MDa).<\/li>\n<\/ul>\n\n<\/div>\n<\/div>\n<\/div>\n\n\n<hr class=\"vf-divider\">\n\n\n<div class=\"embl-grid\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p id=\"af4-saxs\"><\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<article class=\"vf-card vf-card--brand vf-card--bordered vf-u-margin__bottom--800\" default>\n  <div class=\"vf-card__content | vf-stack vf-stack--400\">\n      <h3 class=\"vf-card__heading\">\n      AF4-SAXS    <\/h3>\n              <\/div>\n<\/article>\n\n\n\n\n<figure class=\"vf-figure wp-block-image size-large is-resized\"><img loading=\"lazy\" decoding=\"async\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-2-1024x780.png\" alt=\"\" class=\"wp-image-11655\" width=\"512\" height=\"390\" srcset=\"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-2-1024x780.png 1024w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-2-300x228.png 300w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-2-768x585.png 768w, https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-content\/uploads\/2024\/09\/1d_AF4-SAXS-2.png 1187w\" sizes=\"auto, (max-width: 512px) 100vw, 512px\" \/><figcaption class=\"vf-figure__caption\">Top: Scheme highlighting separation with the AF4 technique. Bottom left: Elution profile emphasizing smaller particles eluting before larger species. Bottom right: Picture of AF4 setup at P12.<\/figcaption><\/figure>\n\n\n\n<p><strong>Asymmetric Flow Field-Flow Fractionation, in line with SAXS, AF4-SAXS<\/strong> features a Postnova AF2000 MultiFlow FFF coupled with the P12 beamline for in situ SAXS measurements. This setup allows for precise fractionation of macromolecules, aggregates, and nanoparticles in the aqueous phase.<\/p>\n\n\n\n<p>A wide variety of sample types, <strong>including proteins, polymers, nanoparticles, and large macromolecular complexes,<\/strong> can be processed, offering unique advantages in separating macromolecules from aggregates or other components that may interfere with SAXS signal analysis.<\/p>\n\n\n\n<p>The sample is injected into the AF4 system using the autosampler, separating the components. After fractionation, The mobile phase goes through the corresponding detectors Postnova Detectors PN3211 UV-Vis for absorbance measurements, PN3609 <strong>Multi-Angle Light Scattering (MALS) <\/strong>for molecular weight determination, and PN3712 <strong>Dynamic Light Scattering (DLS) <\/strong>for hydrodynamic radius measurements, through the elution peaks, providing a comprehensive analysis of the sample.<\/p>\n\n\n\n<p>(Currently a <strong>additional set-up with Wyatt components<\/strong> is under commission and accessible upon request)<\/p>\n\n\n\n<p>Before obtaining beamtime, the <strong>AF4 separation method <\/strong>must be established. This must be <strong>prearranged with bemaline staff.<\/strong><\/p>\n\n\n\n<p>AF4-SAXS experiments typically <strong>require 30-60 min per run<\/strong>, with a recommended exposure time of 0.5 to 2 seconds. <\/p>\n\n\n\n<p>Set-up described in detail here:<\/p>\n\n\n\n<blockquote class=\"vf-blockquote\">\n<p><\/p>\n<p><cite><a href=\"https:\/\/europepmc.org\/abstract\/MED\/37737457\">Quantitative size-resolved characterization of mRNA nanoparticles by in-line coupling of asymmetrical-flow field-flow fractionation with small angle X-ray scattering.<\/a><br>Graewert MA, Wilhelmy C, Bacic T, Schumacher J, Blanchet C, Meier F, Drexel R, Welz R, Kolb B, Bartels K, Nawroth T, Klein T, Svergun D, Langguth P, Haas H<br>Scientific reports,&nbsp;2023<br>doi:10.1038\/s41598-023-42274-z.<\/cite><\/p><\/blockquote>\n\n<\/div>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":21,"featured_media":0,"parent":11465,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"footnotes":""},"embl_taxonomy":[],"class_list":["post-11517","page","type-page","status-publish","hentry"],"acf":[],"embl_taxonomy_terms":[],"_links":{"self":[{"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/pages\/11517","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/users\/21"}],"replies":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/comments?post=11517"}],"version-history":[{"count":63,"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/pages\/11517\/revisions"}],"predecessor-version":[{"id":12091,"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/pages\/11517\/revisions\/12091"}],"up":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/pages\/11465"}],"wp:attachment":[{"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/media?parent=11517"}],"wp:term":[{"taxonomy":"embl_taxonomy","embeddable":true,"href":"https:\/\/www.embl.org\/groups\/small-angle-x-ray-scattering\/wp-json\/wp\/v2\/embl_taxonomy?post=11517"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}