Transcript
SPECTROLIGHTTM 600 IN PLATE
DYNAMIC LIGHT SCATTERING & UV IMAGER Rapidly screen sample quality and observe signs of crystal growth before anything is visible.
Time
Single narrow profile over time - indicates low polydispersity and good stability IDEAL START POINT!
Sub µl Pre-Crystallization Screening: Greatly reduce the amount of protein used and time taken to establish the right buffer, protein concentration and additives for your protein, ensuring a good clean, stable sample. Gain time and move to crystal optimization quickly: Scanning DLS of drops in crystallization plates allows you to rapidly see what is going on in your trials long before anything is visible under a microscope. identify wells that are progressing towards crystallization and eliminate those will precipitate or are crystallization dead ends. Identify new leads and prioritize quickly: Visualize small protein crystals masked by precipitate and differentiate protein from salt using UV imaging.
Slow nucleation - the presence of a slow forming larger species is GOOD!
SpectoLight 600 is a fully automated UV imaging and sub µl in-drop Dynamic Light Scattering system, designed to image, monitor and analyse crystallization, even before anything is visible to the naked eye.
So now you can:
Nucleation too fast – shower of micro crystals at best - NOT OPTIMAL.
Large scale aggregates formed – no crystals! BAD!
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Quickly screen sample stability and quality against buffers, ligands and even detergents prior to crystallization.
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Assess homogeneity, oligomerisation and assembly state.
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Analyse sample behaviour during the crystallization experiment.
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Track each targets progress towards crystallization in-situ and non invasively
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See signs of nucleation long before crystals appear.
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Differentiate protein/other biomolecules from salt crystals.
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Visualize small crystals against a background of precipitate.
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Rapidly record UV and white light images of drops. SpectoLight 600 is manufactured by
Protein crystal hidden by salt crystals!
DLS data for BSA taken under steady state conditions (A) taken directly at the start of crystallization set up and (B) after 12 hours when nuclei can be seen to form.
TAKE THE INTELLIGENT ROUTE TO CRYSTALLIZATION – THE 5 R'S Refine sample quality and stability prior to screening. Reduce the large amount of time wasted on false hits. Recover clear drops - learn vital information which would be missed. Revisit potential hits using in-situ DLS data for guidance. Record, annotate and cross reference all data easily.
SPECTROLIGHTTM 600 TECHNICAL DATA Laserdiode
Wavelength: 660 nm Different wavelength:____________ Optical power: 100 mW, adjustable
Particle size
Measurement range Minimum: 1nm Maximum: approx. 6µm
Sensitivity
Sample concentration with standard laser (660nm) Minimum 0.3mg/ml of lysozyme at 2ul for a ~30kDa protein and 2.0mg/ml for ~14kDa proteins Maximum > 100 mg/ml
Droplet size
About 500nl to 10µl (search algorithm for accurate drop centring)
Detector
For single photon counting Photomultiplier Tube Dark count rate < 300 Hz, Quantum eff.. 5-7% count sensitivity 1.5*105Hz/pW Scattering angle 142° Avalanche photodiode for higher sensitivity wavelengths >660nm
Correlator
Multi-tau architecture correlator to cover a wide sample time range Sample time from 400 ns to 30 s Total 208 quasi logarithmic arranged channels
Dimensions
520 mm x 230 mm x 450mm (LxBxH) Weight: approx. 22kg Power consumption: 115 to 230V 100W
Imaging system
Illumination
White light integrated LED UV by external light source (optional)
5 magnification steps: 0.63, 1.25, 2, 3.2, 6.4 Field of view: 9.4mm, 4.7mm, 3mm, 1.9mm, 0.9mm Resolution(per pixel) - 50µm, 25µm, 16µm, 5µm, 2.5µm CCD Colour Camera - 1280x960 pixels Higher resolution cameras available on request
Operation temperature Temperature of sample Built-in temperature controller- range 4°C to 40°C. (at an ambient temperature of 20°C) Plate Handling
The instrument ejects the plate holder for easy plate access Manual change of plates Plate swapping by robotic handling system Adapted to robot system type: _________________________
Sample format....
Plates in SBS standard crystallization plate format: Sitting Drop: e.g. MRC 96 well or 48 well MAXI plates. Hanging Drop: 24 well SBS plates e.g. Cellstar LCP sandwich plates Terasaki Microbatch Plates (with adaptor) Optional: Customised sample holder__________________________
Software features
Spectrolight 600 software runs on Linux Fully automated plate scanning LIMS database for full data relation and storage Full interactive control of UV and DLS operation Live display of camera image (UV with XtaLight 100) Graphical histogramming software Connection to an external data base (optional)__________________ Connection to third party plate handing system (optional)__________
OTHER MEMBERS OF THE SPECTROLIGHTTM RANGE: X-TALIGHTTM 200 UV/White Light Automated imaging systems
X-TALIGHTTM 100 UV for your existing microscope
SPECTROSIZETM 300 Cuvette DLS system for protein quality control
SPECTROPROBETM400 DLS for your existing microscope
SpectoLight range is manufactured by