ExperimentDynamicLightScattering
ExperimentDynamicLightScattering[Samples]⟹Protocol
Generates a Protocol object to run a Dynamic Light Scattering (DLS) experiment. DLS can be used to measure the size, polydispersity, thermal stability, and attractive/repulsive interactions of particles in solution. Concomitant static light scattering can be used to measure the molecular weight of particles in solution.
ExperimentDynamicLightScattering[Containers]⟹Protocol
Generates a Protocol object to run a Dynamic Light Scattering (DLS) experiment from the samples present in the container. DLS can be used to measure the size, polydispersity, thermal stability, and attractive/repulsive interactions of particles in solution. Concomitant static light scattering can be used to measure the molecular weight of particles in solution. If the Containers and their samples can be analyzed directly with the given assay form factor, no further sample preparation is required.
Dynamic light scattering (DLS) is an analytical technique that correlates changes in scattered light intensity over time to the diffusion rate, and thus the size, of the moving particles in the sample being interrogated. DLS can be used to calculate the hydrodynamic diameter (the diameter of a hypothetical sphere that undergoes diffusion equivalent to that of the relevant particle), size distribution of particles in solution, polydispersity (a measure of the variation of molecular weights of a polymer), isothermal stability, and colloidal stability of particles in solution. Concomitant static light scattering (SLS), which measures the intensity of light scattered relative to the intensity of illumination, allows for the calculation of molar mass. This technique is often used in the formulation development of biologics; DLS measurements can provide insight into whether various formulations will cause aggregation by measuring the weak attractive or repulsive interactions across concentrations. At lower concentrations (>20 mg/mL), the Second Virial Coefficient (B22) and the Diffusion Interaction Parameter (kD) provide this information, while at higher concentrations (>20 mg/mL), this insight into stability is expressed by the Kirkwood-Buff Integral (G22). Thermal stability parameters, including temperature of onset of aggregation (Tagg) and temperatures of onset (Tonset) and midpoint transition temperature (Tm) for global unfolding, can also be extracted from variable-temperature DLS measurements.
Experimental Principles
Figure 1.1: Procedural overview of a light scattering experiment. Step 1: A robotic liquid handler is used to place the input samples into the AssayContainer(s) either directly or via SampleLoadingPlate (if the samples are not already contained in containers compatible with the given AssayFormFactor), and perform any dilutions or serial dilutions, if applicable. Step 2: The AssayContainer(s) are incubated at the Temperature for the EquilibrationTime before any measurements are made. Step 3: Light scattering measurements are made. Process A, Step 4: Dynamic light scattering data is acquired as speckle patterns. Step 5: An autocorrelation function is generated based on the plot of intensity versus time. Step 6: The autocorrelation function is fit via a regularization algorithm. Step 7: Data including hydrodynamic radius, size distribution, polydispersity index, B22, kD, Tagg, Tm are obtained. Process B, Step 4: Static light scattering intensity data is generated as counts per second. Step 5: Data including molar mass, A2 (or B22), and Tagg are obtained. Process C, Step 4: When AssayFormFactor is Plate, well images are acquired.
Figure 1.2: In a dynamic light scattering experiment, a laser is used to probe particles in solution, which scatter the incoming light, thereby generating speckle patterns. A series of speckle patterns are generated at discrete time intervals. Small particles exhibit faster Brownian motion than large particles, so the intensity of the scattered light varies more between time intervals for small particles than for large particles. An autocorrelation function can be generated for the variation of intensity with time, and the distribution of the hydrodynamic radii of the particles may be obtained from the autocorrelation function via the Stokes-Einstein equation.
Figure 1.3: In a static light scattering experiment, a laser is used to probe particles in solution which scatter the incoming light. The intensity yields the Rayleigh ratio of the particles in solution, which, in conjunction with the concentration and refractive index of the solvent, may be used to obtain quantitative analysis of the molecular weight of the analyte when it is probed at a range of concentrations.
Figure 1.4: Dynamic and static light scattering may be used to probe the isothermal stability of a sample. After an initial equilibration time at a given temperature, light scattering data is collected. Data collection occurs over the length of the experiment separated by a measurement delay time, and sizing data can be plotted over a range of times.
Figure 1.5: Dynamic and static light scattering may be used to probe the colloidal stability of a sample. Samples are diluted to a range of concentrations, and light scattering data collected at each concentration yields data expressing the weak attractive or repulsive interactions across concentrations. At lower concentrations (>20 mg/mL), the Second Virial Coefficient (B22) and the Diffusion Interaction Parameter (kD) provide this information, while at higher concentrations (>20 mg/mL), this insight into stability is expressed by the Kirkwood-Buff Integral (G22). When a linearized avalanche photodiode is employed, static light scattering across the range of concentrations also affords the precise molecular weight of the analyte.
Figure 1.6: Dynamic and static light scattering may be used to obtain a melting curve for an analyte. Samples are interrogated at a range of temperatures. The plot of hydrodynamic radius or static light scattering intensity versus temperature can be used to obtain thermal stability parameters, including temperature of onset of aggregation (Tagg) and temperatures of onset (Tonset) and midpoint transition temperature (Tm) for global unfolding.
Instrumentation
Uncle
Figure 2.1.1: The Uncle is a multimode spectrophotometer with the capacity to interrogate forty eight samples in one run. Samples must be assayed in Uncle compatible capillary strips. Each strip contains 16 quartz capillaries and each capillary requires 9 uL of sample. Sample dynamic light scattering can be quantified by a 660 nm laser diode and detected using an avalanche photodiode module. Laser power can be specified from opaque to full power (0 - 100% respectively). For all measurements, each capillary is interrogated independently. Capillary strip temperature is controlled by a thermoelectric heating and cooling plate and can range from 15°C to 95°C.
DynaPro
Figure 2.2.1: The DynaPro is a plate reader with the capacity to execute Dynamic Light Scattering (DLS) or Static Light Scattering (SLS) experiments on up to 384 samples in one run. Samples may be assayed in 96- or 384-well plates. Both DLS and SLS can be quantified by an 830 nm laser diode and detected using a linearized avalanche photodiode module. Laser power can be specified from opaque to full power (10 - 100% respectively). For all measurements, each well is interrogated independently. The temperature of the incubation chamber is controlled by a heating element and can range from 4°C to 85°C.
Experiment Options
General
AssayType
The Dynamic Light Scattering (DLS) assay that is run. SizingPolydispersity makes a single DLS measurement that provides information about the size and polydispersity (defined as the ratio of mass-average molar mass to number-average molar mass) of particles in the input samples. ColloidalStability makes DLS measurements at various dilutions of a sample below 25 mg/mL to calculate the diffusion interaction parameter (kD) and the second virial coefficient (B22 or A2), and does the same for a sample of mass concentration 20-100 mg/mL to calculate the Kirkwood-Buff Integral (G22) at each dilution concentration; Static Light Scattering (SLS) measurements can be used to calculate A2 and the molecular weight of the analyte. MeltingCurve makes DLS measurements over a range of temperatures in order to calculate melting temperature (Tm), temperature of aggregation (Tagg), and temperature of onset of unfolding (Tonset). IsothermalStability makes multiple DLS measurements at a single temperature over time in order to probe stability of the analyte at a particular temperature.
Default Calculation: The AssayType is set to IsothermalStability if any of the options in the "Isothermal Stability" category are specified, to MeltingCurve if any of the options in the "Melting Curve" category are defined, to ColloidalStability if Dilution-related options are specified, and to SizingPolydispersity otherwise.
Pattern Description: SizingPolydispersity, IsothermalStability, MeltingCurve, or ColloidalStability.
AssayFormFactor
Indicates if the sample is loaded in capillary strips, which are utilized by a Multimode Spectrophotometer, or a standard plate which is utilized by a DLS Plate Reader.
Default Calculation: AssayFormFactor is set to Capillary if the Instrument model is set to Model[Instrument,MultimodeSpectrophotometer,"Uncle"] and Plate if Instrument is set to Model[Instrument,DLSPlateReader,"DynaPro"]. If neither AssayFormFactor nor Instrument is defined, AssayFormFactor is set to Capillary if SampleVolume is set to 25 uL and Object[Instrument,DLSPlateReader,"DynaPro" if SampleVolume is set to greater than 25 uL.
Instrument
The instrument used for this experiment. Options comprise Model[Instrument,MultimodeSpectrophotometer,"Uncle"], or any instruments of that model, which uses a capillary form factor and can perform fluorescence as well as light scattering experiments assays, and Model[Instrument,DLSPlateReader,"DynaPro"], or any instruments of that model, which uses a plate form factor and performs only light scattering experiments.
Default Calculation: Instrument is set to Model[Instrument,MultimodeSpectrophotometer,"Uncle"] if AssayFormFactor is set to Capillary and Model[Instrument,DLSPlateReader,"DynaPro"] if AssayFormFactor is set to Plate. If neither AssayFormFactor nor Instrument is defined, Instrument is set to Model[Instrument,MultimodeSpectrophotometer,"Uncle"] if SampleVolume is set to 25 uL or less and Model[Instrument,DLSPlateReader,"DynaPro"] if SampleVolume is set to greater than 25 uL.
Pattern Description: An object of type or subtype Object[Instrument, MultimodeSpectrophotometer], Model[Instrument, MultimodeSpectrophotometer], Object[Instrument, DLSPlateReader], or Model[Instrument, DLSPlateReader]
Programmatic Pattern: ObjectP[{Object[Instrument, MultimodeSpectrophotometer], Model[Instrument, MultimodeSpectrophotometer], Object[Instrument, DLSPlateReader], Model[Instrument, DLSPlateReader]}] | Automatic
AssayContainers
The capillary strips or plates that the samples in this experiment are assayed in. Options comprise a list of one of Model[Container, Plate, "96 well Flat Bottom DLS Plate"], Model[Container, Plate, "96 well Clear Flat Bottom DLS Plate"], Model[Container, Plate, "384-well Aurora Flat Bottom DLS Plate"], each of which use a plate form factor, and a list of one, two, or three of Model[Container, Plate, CapillaryStrip, "Uncle 16-capillary strip"], which uses a capillary form factor, or any objects of those models. If AssayFormFactor is Capillary, it is strongly recommended to allow AssayContainers to resolve automatically.
Default Calculation: When AssayFormFactor is set to Plate, AssayContainers is set to {Model[Container, Plate, "96 well Flat Bottom DLS Plate"]} if SampleVolume is set to more than 30 Microliter and there are fewer than 96 wells required for the assay, and to {Model[Container, Plate, "384-well Aurora Flat Bottom DLS Plate"]} otherwise. When AssayFormFactor is set to Capillary, AssayContainers is set to one or more instances of Model[Container, Plate, CapillaryStrip, "Uncle 16-capillary strip"], depending on the number of wells required for the assay.
Pattern Description: List of one or more an object of type or subtype Object[Container, Plate] or Model[Container, Plate] or a prepared sample entries.
Programmatic Pattern: {(ObjectP[{Object[Container, Plate], Model[Container, Plate]}] | _String)..} | Automatic
Sample Loading
SampleLoadingPlate
The container into which input samples are transferred (or in which input sample dilutions are performed when AssayType is ColloidalStability) before centrifugation and transfer into the AssayContainer(s) for DLS measurement.
Default Calculation: The SampleLoadingPlate is set to Null when AssayFormFactor is Plate and to Model[Container, Plate, "96-well PCR Plate"] when AssayFormFactor is Capillary.
Pattern Description: An object of type or subtype Object[Container, Plate] or Model[Container, Plate] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Object[Container, Plate], Model[Container, Plate]}] | _String) | Automatic) | Null
NumberOfReplicates
The number of times the specified Dynamic Light Scattering (DLS) assay is run on each input sample. In practice, this refers to the number of wells of the AssayContainer(s) that each input sample occupies. For example, ExperimentDynamicLightScattering[{input1, input2}, NumberOfReplicates->2] is equivalent to ExperimentDynamicLightScattering[{input1, input1, input2, input2}]. When AssayType is SizingPolydispersity, IsothermalStability, and MeltingCurve, NumberOfReplicates refers to the number of wells for each distinct input sample. When AssayType is ColloidalStability, NumberOfReplicates refers to the replicates of each dilution concentration of each sample. At least two replicates of each dilution concentration are required for ColloidalStability, and three are recommended.
Pattern Description: Greater than or equal to 1 and less than or equal to 384 in increments of 1 or Null.
AssayContainerFillDirection
WhenAssayFormFactor is Capillary, controls how the capillary strip AssayContainers are filled. Column indicates that all 16 wells of an AssayContainer capillary strip (Model[Container, Plate, CapillaryStrip,"Uncle 16-capillary strip"]) are filled with input samples or sample dilutions before starting to fill a second capillary strip (up to 3 strips, 48 wells). Row indicates that one well of each capillary strip is filled with input samples or sample dilutions before filling the second well of each strip. Setting the AssayContainerFillDirection to Column will minimize the number of capillary strips needed for the experiment, while Row will always use three capillary strips if there are more than two input samples. Setting this option to Row will ensure that replicate dilution concentration measurements in ColloidalStability occur in separate capillary strips. When AssayFormFactor is Plate, controls the direction the AssayContainer is filled: either Row, Column, SerpentineRow, or SerpentineColumn.
SampleVolume
When AssayType is SizingPolydispersity, MeltingCurve, or IsothermalStability, the SampleVolume is the amount of each input sample that is transferred into the SamplePreparationPlate before the SamplePreparationPlate is centrifuged and 10 uL of each sample is transferred into the AssayContainer(s) for DLS measurement. When the AssayType is ColloidalStability, the amount of input sample required for the experiment is specified with either the DilutionCurve or SerialDilutionCurve option.
Default Calculation: If the AssayType is SizingPolydispersity, MeltingCurve, or IsothermalStability, when the AssayFormFactor is Capillary, SampleVolume is set to 15 uL; when the AssayFormFactor is Plate, SampleVolume is set to 30 uL (for a 384-well plate) or 100 uL (for a 96-well plate); and set to Null otherwise.
Pattern Description: Greater than or equal to 15 microliters and less than or equal to 100 microliters or Null.
Temperature
Pattern Description: Greater than or equal to 4 degrees Celsius and less than or equal to 95 degrees Celsius.
EquilibrationTime
The length of time for which the samples are held in the chamber which is incubated at the Temperature before the first DLS measurement is made, in order to warm or cool the samples to Temperature.
CapillaryLoading
The loading method for capillaries. When set to Robotic, capillaries are loaded by liquid handler. When set to Manual, capillaries are loaded by a multichannel pipette. Each method presents specific mechanical challenges due to the difficulty of precise loading.
Default Calculation: When AssayFormFactor is Capillary, set to Manual. When AssayFormFactor is Plate, set to Null.
WellCover
When AssayFormFactor is Plate, determines what covering will be used for wells. Available coverings are plate seal, and oil; other covers (e.g. lids) have not yet been evaluated for their effects on light scattering optics.
Default Calculation: When AssayFormFactor is Plate, automatically set to Model[Sample,"Silicone Oil"]. When AssayFormFactor is Capillary, automatically set to Null.
Pattern Description: An object of type or subtype Object[Item, PlateSeal], Model[Item, PlateSeal], Object[Sample], or Model[Sample] or a prepared sample or None or Null.
Programmatic Pattern: (((ObjectP[{Object[Item, PlateSeal], Model[Item, PlateSeal], Object[Sample], Model[Sample]}] | _String) | None) | Automatic) | Null
WellCoverHeating
When WellCover is Model[Item,PlateSeal] or any object of that model, indicates if the plate seal is heated.
Default Calculation: When AssayFormFactor is Plate and WellCover is set to a Model[Item,PlateSeal] or Object[Item,PlateSeal], automatically set to True. When AssayFormFactor is Capillary or WellCover is set to anything other than Model[Item,PlateSeal] or Object[Item,PlateSeal], automatically set to Null.
Sample Dilution
ReplicateDilutionCurve
Indicates if a NumberOfReplicates number of StandardDilutionCurves or SerialDilutionCurves are made for each input sample. When ReplicateDilutionCurve is True, the replicate DLS measurements for ColloidalStability assay are made from the same concentration of each of the StandardDilutionCurves or SerialDilutionCurves created from a given input sample. When ReplicateDilutionCurve is False, the replicate DLS measurements for the ColloidalStability assay are made from aliquots of a given concentration of the DilutionCurve or SerialDilutionCurve.
Default Calculation: If the AssayType is ColloidalStability, ReplicateDilutionCurve is set to True if the AssayFormFactor is Plate, and to False if AssayFormFactor is Capillary. If the AssayType is not ColloidalStability, ReplicateDilutionCurve is set to Null.
StandardDilutionCurve
The collection of dilutions that are performed on each input sample in the SampleLoadingPlate or AssayContainer. For Fixed Dilution Volume Dilution Curves, the Sample Amount is the volume of the sample that is mixed with the Buffer to the Total Volume to create a desired concentration. For Fixed Target Concentration Dilution Curves, the Target Concentration is the desired final concentration of analyte after dilution of the input samples with the Buffer.
Figure 3.1: Use the StandardDilutionCurve option to create a collection of dilutions that will be performed on the sample.
Default Calculation: The Dilution is set to Null if the AssayType is SizingPolydispersity, IsothermalStability, or MeltingCurve, or if the SerialDilutionCurve option is specified. Otherwise, when AssayFormFactor is Capillary, the option is set to be {{14 uL, 10 mg/mL},{14 uL, 8 mg/mL},{14 uL, 6 mg/mL},{14 uL, 4 mg/mL},{14 uL, 2 mg/mL}} if ReplicateDilutionCurve is True, or to be {{14 uL times NumberOfReplicates), 10 mg/mL},{14 uL times NumberOfReplicates), 8 mg/mL},{14 uL times NumberOfReplicates), 6 mg/mL},{14 uL times NumberOfReplicates), 4 mg/mL},{14 uL times NumberOfReplicates), 2 mg/mL}} if ReplicateDilutionCurve is False. When AssayFormFactor is Plate, the Sample Amount is set to a series of dilutions of 90 uL or 30 uL (depending on number of wells).
Pattern Description: List of one or more {Sample Amount, Target Concentration} entries or list of one or more {Sample Amount, Total Volume} entries or Null.
Programmatic Pattern: (({{RangeP[0*Microliter, $MaxTransferVolume], RangeP[0*Microliter, $MaxTransferVolume]}..} | {{RangeP[0*Microliter, $MaxTransferVolume], GreaterEqualP[0*(Milligram/Milliliter)] | GreaterP[0*Molar]}..}) | Automatic) | Null
SerialDilutionCurve
The collection of dilutions that are performed on each input sample in the SampleLoadingPlate or AssayContainer. For Serial Dilution Factors, the sample will be diluted with the Buffer by the dilution factor at each transfer step. For example, a SerialDilutionCurve of {36*Microliter,{1,1.25,2,2},1} for a 100 mg/mL sample will result in 4 dilutions with concentrations of 100 mg/mL, 80 mg/mL, 40 mg/mL, and 20 mg/mL. For Serial Dilution Volumes, the Transfer Volume is taken out of the sample and added to a second well with the Buffer Volume of the Buffer. It is mixed, then the Transfer Volume is taken out of that well to be added to a third well. This is repeated to make Number Of Dilutions diluted samples. For example, if a 100 mg/mL sample with a Transfer Volume of 30 Microliters, a Buffer Volume of 10 Microliters and a Number of Dilutions of 2 is used, it will create a SerialDilutionCurve of 75 mg/mL, 56.3 mg/mL, and 42.2 mg/mL.
Figure 3.2: Use the SerialDilutionCurve option to create a collection of serial dilutions that will be performed on the sample.
Default Calculation: The SerialDilutionCurve is set to Null if the AssayType is SizingPolydispersity, IsothermalStability, or MeltingCurve, or if the StandardDilutionCurve option is specified. Otherwise, when AssayFormFactor is Capillary, the option is set to be {15 uL, {1, 1.25, 1.333, 1.5, 2}, 1} if ReplicateDilutionCurve is True, or to be {(15 uL times NumberOfReplicates), {1, 1.25, 1.333, 1.5, 2}, 1} if ReplicateDilutionCurve is False. When AssayFormFactor is Plate, the Sample Volume is set to 90 uL or 30 uL (depending on number of wells). In both cases, the dilution factors with respect to the input sample are {1, 1.25, 1.333, 1.5, 2}, so a 10 mg/mL sample would be diluted to {10 mg/mL, 8 mg/mL, 6 mg/mL, 4 mg/mL, 2 mg/mL}.
Programmatic Pattern: (({RangeP[0*Microliter, $MaxTransferVolume], RangeP[0*Microliter, $MaxTransferVolume], RangeP[1, 48, 1]} | {RangeP[0*Microliter, $MaxTransferVolume], {GreaterEqualP[1]..}, GreaterEqualP[1, 1]}) | Automatic) | Null
DilutionMixType
Default Calculation: Automatically set to Manual when AssayType is ColloidalStability, and to Null otherwise.
DilutionNumberOfMixes
The number of pipette out and in cycles that is used to mix the sample with the Buffer to make the DilutionCurve.
Default Calculation: Automatically set to Null if the AssayType is SizingPolydispersity, IsothermalStability, or MeltingCurve; or if either the DilutionNumberOfMixes or DilutionMixRate is specified as Null; or if MixFormFactor is set to Vortex; and to 5 otherwise.
Pattern Description: Greater than or equal to 0 and less than or equal to 20 in increments of 1 or Null.
DilutionMixRate
The speed at which the dilution sample is pipetted out of and into the dilution to mix the sample with the Buffer to make the DilutionCurve.
Default Calculation: Automatically set to Null if the AssayType is SizingPolydispersity, IsothermalStability, or MeltingCurve; or if either the DilutionNumberOfMixes or DilutionMixRate is specified as Null; or if MixFormFactor is set to Vortex; and to 30 uL/second otherwise.
Pattern Description: Greater than or equal to 0.4 microliters per second and less than or equal to 250 microliters per second or Null.
DilutionMixInstrument
The instrument used to mix the dilutions in the SampleLoadingPlate or AssayContainer used for dilution.
Default Calculation: DilutionMixInstrument is automatically set to Model[Instrument,Pipette] if DilutionMixType is set to Pipette; to Model[Instrument,Vortex] if DilutionMixType is set to Vortex; to Model[Instrument,Nutator] if DilutionMixType is set to Nutator; and to Null if AssayType is SizingPolydispersity, IsothermalStability, or MeltingCurve.
Pattern Description: An object of type or subtype Model[Instrument, Pipette], Object[Instrument, Pipette], Model[Instrument, Vortex], Object[Instrument, Vortex], Model[Instrument, Nutator], or Object[Instrument, Nutator] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, Pipette], Object[Instrument, Pipette], Model[Instrument, Vortex], Object[Instrument, Vortex], Model[Instrument, Nutator], Object[Instrument, Nutator]}] | Automatic) | Null
Sample and Solvent Information
SolventName
For each input sample, the name of the solvent specified in the DynaPro software. This option is only necessary if the AssayFormFactor is Plate and either CollectStaticLightScattering is True or the AssayType is ColloidalStability.
Default Calculation: If the AssayFormFactor is Plate and either CollectStaticLightScattering is True or the AssayType is ColloidalStability, SolventName is set based on SolventViscosity and SolventRefractiveIndex if those are specified, set to "Water" if the Buffer is "Water", or to "New" if otherwise unspecified.
Pattern Description: New or {Water, Acetic Acid 0.5%, Acetic Acid 1%, Acetic Acid 2%, Acetic Acid 3%, Acetic Acid 4%, Acetic Acid 5%, Acetic Acid 6%, Acetic Acid 7%, Acetic Acid 8%, Acetic Acid 9%, Acetone 0.5%, Acetone 1.%, Acetone 1.5%, Acetone 2.%, Acetone 2.5%, Acetone 3.%, Acetone 3.5%, Acetone 4.%, Acetone 4.5%, Acetone 5.%, Acetone 6%, Acetone 100%, Ammonium Chloride 0.5%, Ammonium Chloride 1%, Ammonium Chloride 2%, Ammonium Chloride 3%, Ammonium Chloride 4%, Ammonium Chloride 5%, Ammonium Chloride 6%, Ammonium Chloride 7%, Ammonium Chloride 8%, Ammonium Chloride 9%, Ammonium Chloride 10%, Calcium Chloride 0.5%, Calcium Chloride 1%, Calcium Chloride 2%, Calcium Chloride 3%, Calcium Chloride 4%, Calcium Chloride 5%, Calcium Chloride 6%, Calcium Chloride 7%, Calcium Chloride 8%, Calcium Chloride 9%, Calcium Chloride 10%, Cyclohexane, Ethanol 1%, Ethanol 2%, Ethanol 3%, Ethanol 4%, Ethanol 5%, Ethanol 6%, Ethanol 7%, Ethanol 8%, Ethanol 9%, Ethanol 10%, Ethanol 15%, Ethanol 20%, Ethanol 100%, Ethylene Glycol 1%, Ethylene Glycol 2%, Ethylene Glycol 3%, Ethylene Glycol 4%, Ethylene Glycol 5%, Ethylene Glycol 6%, Ethylene Glycol 7%, Ethylene Glycol 8%, Ethylene Glycol 9%, Ethylene Glycol 10%, Ethylene Glycol 20%, Glucose 1%, Glucose 2%, Glucose 3%, Glucose 4%, Glucose 5%, Glucose 6%, Glucose 7%, Glucose 8%, Glucose 9%, Glucose 10%, Glucose 20%, Glycerol 1%, Glycerol 2%, Glycerol 3%, Glycerol 4%, Glycerol 5%, Glycerol 6%, Glycerol 7%, Glycerol 8%, Glycerol 9%, Glycerol 10%, Glycerol 15%, Glycerol 20%, Glycerol 25%, Glycerol 30%, Glycerol 40%, Glycerol 50%, Glycerol 100%, Hexane, Lithium Chloride 0.5%, Lithium Chloride 1%, Lithium Chloride 2%, Lithium Chloride 3%, Lithium Chloride 4%, Lithium Chloride 5%, Lithium Chloride 6%, Lithium Chloride 7%, Lithium Chloride 8%, Lithium Chloride 9%, Lithium Chloride 10%, Magnesium Chloride 0.5%, Magnesium Chloride 1%, Magnesium Chloride 2%, Magnesium Chloride 3%, Magnesium Chloride 4%, Magnesium Chloride 5%, Magnesium Chloride 6%, Magnesium Chloride 7%, Magnesium Chloride 8%, Magnesium Chloride 9%, Magnesium Chloride 10%, Maltose 0.5%, Maltose 1%, Maltose 2%, Maltose 3%, Maltose 4%, Maltose 5%, Maltose 6%, Maltose 7%, Maltose 8%, Maltose 9%, Maltose 10%, Methanol 1%, Methanol 2%, Methanol 3%, Methanol 4%, Methanol 5%, Methanol 6%, Methanol 7%, Methanol 8%, Methanol 9%, Methanol 10%, Methanol 15%, Methanol 20%, Methanol 100%, Oleic Acid, PBS, Sodium Acetate 0.5%, Sodium Acetate 1%, Sodium Acetate 2%, Sodium Acetate 3%, Sodium Acetate 4%, Sodium Acetate 5%, Sodium Acetate 6%, Sodium Acetate 7%, Sodium Acetate 8%, Sodium Acetate 9%, Sodium Acetate 10%, Sodium Bromide 0.5%, Sodium Bromide 1%, Sodium Bromide 2%, Sodium Bromide 3%, Sodium Bromide 4%, Sodium Bromide 5%, Sodium Bromide 6%, Sodium Bromide 7%, Sodium Bromide 8%, Sodium Bromide 9%, Sodium Bromide 10%, Sodium Chloride 0.1%, Sodium Chloride 0.2%, Sodium Chloride 0.3%, Sodium Chloride 0.4%, Sodium Chloride 0.5%, Sodium Chloride 1.%, Sodium Chloride 1.5%, Sodium Chloride 2.%, Sodium Chloride 2.5%, Sodium Chloride 3.%, Sodium Chloride 3.5%, Sodium Chloride 4.%, Sodium Chloride 4.5%, Sodium Chloride 5.%, Sodium Chloride 6%, Sodium Chloride 7%, Sodium Chloride 8%, Sodium Chloride 9%, Sodium Chloride 10%, Sodium Chloride 15%, Sodium Chloride 20%, Sodium Hydroxide 0.5%, Sodium Hydroxide 1%, Sodium Hydroxide 2%, Sodium Hydroxide 3%, Sodium Hydroxide 4%, Sodium Hydroxide 5%, Sodium Hydroxide 6%, Sodium Hydroxide 7%, Sodium Hydroxide 8%, Sodium Hydroxide 9%, Sodium Hydroxide 10%, Sodium Nitrate 0.5%, Sodium Nitrate 1%, Sodium Nitrate 2%, Sodium Nitrate 3%, Sodium Nitrate 4%, Sodium Nitrate 5%, Sodium Nitrate 6%, Sodium Nitrate 7%, Sodium Nitrate 8%, Sodium Nitrate 9%, Sodium Nitrate 10%, Sodium Sulfate 0.5%, Sodium Sulfate 1%, Sodium Sulfate 2%, Sodium Sulfate 3%, Sodium Sulfate 4%, Sodium Sulfate 5%, Sodium Sulfate 6%, Sodium Sulfate 7%, Sodium Sulfate 8%, Sodium Sulfate 9%, Sodium Sulfate 10%, Sucrose 1%, Sucrose 2%, Sucrose 3%, Sucrose 4%, Sucrose 5%, Sucrose 6%, Sucrose 7%, Sucrose 8%, Sucrose 9%, Sucrose 10%, Sucrose 15%, Sucrose 20%, Toluene, Urea 1%, Urea 5%, Urea 6%, Urea 7%, Urea 8%, Urea 9%, Urea 10%, Urea 15%, Urea 20%} or Null.
SolventViscosity
For each input sample, the viscosity of the solvent specified in the DynaPro software. This option is only necessary if the AssayFormFactor is Plate and either CollectStaticLightScattering is True or the AssayType is ColloidalStability.
Default Calculation: If the AssayFormFactor is Plate and either CollectStaticLightScattering is True or the AssayType is ColloidalStability, SolventViscosity is set based on SolventName if it is specified or resolved, as described in $DLSSolventLookupTable, and set to Null otherwise.
Pattern Description: Greater than or equal to 0 centipoise and less than or equal to 50 centipoise or Null.
SolventRefractiveIndex
For each input sample, the viscosity of the solvent specified in the DynaPro software. This option is only necessary if the AssayFormFactor is Plate and either CollectStaticLightScattering is True or the AssayType is ColloidalStability.
Default Calculation: If the AssayFormFactor is Plate and either CollectStaticLightScattering is True or the AssayType is ColloidalStability, SolventRefractiveIndex is set based on SolventName if it is specified or resolved, as described in $DLSSolventLookupTable, and set to Null otherwise.
Analyte
For each input sample, the Model[Molecule] member of the Composition field whose concentration is used to collect static light scattering data, or to calculate B22/A2 and kD or G22 when the AssayType is ColloidalStability.
Default Calculation: Automatically set to Null when CollectStaticLightScattering is False or AssayFormFactor is Capillary and the AssayType is not ColloidalStability. When AssayType is ColloidalStability, or when CollectStaticLightScattering is True, automatically set to the first value in the Analytes field of the input sample, or, if not populated, to the first analyte in the Composition field of the input sample, or if none exist, the first identity model of any kind in the Composition field.
AnalyteMassConcentration
For each input sample, the initial mass concentration of the Analyte before any dilutions outlined by the StandardDilutionCurve or SerialDilutionCurve are performed when the AssayType is ColloidalStability.
Default Calculation: Automatically set to Null when CollectStaticLightScattering is False or AssayFormFactor is Capillary and the AssayType is not ColloidalStability. When AssayType is ColloidalStability, or when CollectStaticLightScattering is True, automatically set to the mass concentration of the first value in the Analytes field of the input sample, or, if not populated, to that of the first analyte in the Composition field of the input sample, or if none exist, that of the first identity model of any kind in the Composition field.
Pattern Description: Greater than or equal to 0 milligrams per milliliter or greater than 0 molar or Null.
Programmatic Pattern: ((GreaterEqualP[0*(Milligram/Milliliter)] | GreaterP[0*Molar]) | Automatic) | Null
AnalyteRefractiveIndexIncrement
For each input sample, the known or estimated refractive index increment (also known as dn/dc) of the Analyte.
Default Calculation: Automatically set to Null when CollectStaticLightScattering is False or AssayFormFactor is Capillary and the AssayType is not ColloidalStability. When AssayType is ColloidalStability, or when CollectStaticLightScattering is True and AssayFormFactor is Plate, automatically set to 0.185 Milliliter/Gram if the Analyte is a Model[Molecule, Protein] or Model[Molecule, cDNA], and to 0.15 Milliliter/Gram if the Analyte is a Model[Molecule, Polymer] or Model[Molecule, Oligomer].
Buffer
The buffer or solvent that is used to dilute the sample to make a DilutionCurve or in collecting static light scattering.
Default Calculation: The Buffer is automatically set to Null when CollectStaticLightScattering is False or AssayFormFactor is Capillary and the AssayType is not ColloidalStability. When AssayType is ColloidalStability, or when CollectStaticLightScattering is True, the Buffer is set to the BlankBuffer if it is specified, to the Solvent of the input sample if the BlankBuffer is not specified, and to Model[Sample, "Milli-Q water"] otherwise.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankBuffer
The sample that is used as a 0 mg/mL blank in ColloidalStability assays or when CollectStaticLightScattering is True, to determine the diffusion coefficient at infinite dilution.
Default Calculation: The BlankBuffer is set to the Buffer if the AssayType is ColloidalStability or if CollectStaticLightScattering is True and to Null otherwise.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Colloidal Stability
CalculateMolecularWeight
When AssayFormFactor is Plate, determines if Static Light Scattering (SLS) is used to calculate weight-average molecular weight.
Default Calculation: When AssayFormFactor is Plate and AssayType is ColloidalStability, automatically set to True. when CollectStaticLightScattering is False or AssayFormFactor is Capillary or AssayFormFactor is Capillary, automatically set to Null.
Light Scattering
CollectStaticLightScattering
Indicates whether static light scattering (SLS) data are collected along with DLS data. Reliable static light scattering data can be obtained only if the combination of plate and solvent has previously been calibrated.
CalibratePlate
When AssayFormFactor is Plate, indicates whether the AssayContainer should be calibrated by measuring the scattered light intensities of a series of dilutions of a standard sample before any other data collection.
Default Calculation: When AssayFormFactor is Plate and CollectStaticLightScattering is True, automatically set to True.
NumberOfAcquisitions
For each Dynamic Light Scattering (DLS) measurement, the number of series of speckle patterns that are each collected over the AcquisitionTime to create the measurement's autocorrelation curve.
AcquisitionTime
For each Dynamic Light Scattering (DLS) measurement, the length of time that each acquisition generates speckle patterns to create the measurement's autocorrelation curve.
AutomaticLaserSettings
Indicates if the LaserPower and DiodeAttenuation are automatically set at the beginning of the assay by the Instrument to levels ideal for the samples, such that the count rate falls within an optimal, predetermined range.
Default Calculation: Automatically set to False if either LaserPower or DiodeAttenuation is not Null, and to True otherwise.
LaserPower
The Percent of the max laser power that is used to make Dynamic Light Scattering (DLS) measurements. The laser level is optimized at run time by the instrument software when AutomaticLaserSettings is True and LaserLevel is Null.
Default Calculation: The LaserPower option is set to 100Percent if AutomaticLaserSettings is False and to Null otherwise.
Pattern Description: Greater than or equal to 1 percent and less than or equal to 100 percent or Null.
DiodeAttenuation
The Percent of scattered signal that is allowed to reach the avalanche photodiode (in Capillary-type assays) or prevented from reaching the avalanche photodiode (in Plate-type assays). The attenuator level is optimized at run time by the instrument software when AutomaticLaserSettings is True and DiodeAttenuation is Null.
Default Calculation: If AutomaticLaserSettings is False and AssayFormFactor is Plate, the DiodeAttenuation option is set to 0Percent; if AutomaticLaserSettings is False and AssayFormFactor is Capillary, the DiodeAttenuation option is set to 100Percent, and to Null in all other cases.
Pattern Description: Greater than or equal to 0 percent and less than or equal to 100 percent or Null.
Isothermal Stability
MeasurementDelayTime
The length of time between the consecutive Dynamic Light Scattering (DLS) measurements for a specific AssayContainer well during an IsothermalStability assay. The duration of the experiment is set either by this option or by the total IsothermalRunTime.
Default Calculation: When AssayType is IsothermalStability and when AssayFormFactor is Capillary, the MeasurementDelayTime option is set to a value calculated by the Instrument manufacture's proprietary algorithm (dependent on the number of input samples and the IsothermalAttenuatorAdjustment); when AssayFormFactor is Plate, MeasurementDelayTime is automatically set to 1 hour. When AssayType is SizingPolydispersity, MeltingCurve, or ColloidalStability, automatically set to Null.
Pattern Description: Greater than or equal to 70 seconds and less than or equal to 3.75 hours or Null.
IsothermalMeasurements
The number of separate DLS measurements that are made during the IsothermalStability assay, either separated by MeasurementDelayTime or distributed over IsothermalRunTime.
Default Calculation: The IsothermalMeasurements option is set to 10 if the AssayType is IsothermalStability and the IsothermalRunTime is Null, or to Null otherwise.
Pattern Description: Greater than or equal to 0 and less than or equal to 8022 in increments of 1 or Null.
IsothermalRunTime
The total length of the IsothermalStability assay during which the IsothermalMeasurements number of Dynamic Light Scattering (DLS) measurements are made. The duration of the experiment is set either by this option or by the MeasurementDelayTime.
Default Calculation: The IsothermalRunTime is set to 10 times the MeasurementDelayTime if the AssayType is IsothermalStability and the IsothermalMeasurements option is Null, or to Null otherwise.
Pattern Description: Greater than or equal to 70 seconds and less than or equal to 72 hours or Null.
IsothermalAttenuatorAdjustment
Indicates if the attenuator level is automatically set for each DLS measurement throughout the IsothermalStability assay. If First, the attenuator level is automatically set for the first DLS measurement and the same level is used throughout the assay.
Default Calculation: The IsothermalAttenuatorAdjustment is set to Every if the AssayType is IsothermalStability and to Null otherwise.
Melting Curve
MinTemperature
The low temperature of the heating or cooling curve; the starting temperature when TemperatureRampOrder is {Heating,Cooling}.
Default Calculation: When AssayType is MeltingCurve, automatically set to 4 Celsius if AssayFormFactor is Plate and to 15 Celsius when AssayFormFactor is Capillary. When AssayType is SizingPolydispersity, IsothermalStability, or ColloidalStability, automatically set to Null.
Pattern Description: Greater than or equal to 4 degrees Celsius and less than or equal to 100 degrees Celsius or Null.
MaxTemperature
The high temperature of the heating or cooling curve; the starting temperature when TemperatureRampOrder is {Cooling,Heating}.
Default Calculation: When AssayType is MeltingCurve, automatically set to 85 Celsius if AssayFormFactor is Plate and 95 Celsius if AssayFormFactor is Capillary. When AssayType is SizingPolydispersity, IsothermalStability, or ColloidalStability, automatically set to Null.
Pattern Description: Greater than or equal to 4 degrees Celsius and less than or equal to 100 degrees Celsius or Null.
TemperatureRampOrder
The order of temperature ramping (i.e., heating followed by cooling or vice versa) to be performed in each cycle. Heating is defined as going from MinTemperature to MaxTemperature; cooling is defined as going from MaxTemperature to MinTemperature.
Default Calculation: When AssayType is MeltingCurve, automatically set to {Heating,Cooling}. When AssayType is SizingPolydispersity, IsothermalStability, or ColloidalStability, automatically set to Null.
NumberOfCycles
Default Calculation: When AssayType is MeltingCurve, automatically set to 1 cycle. When AssayType is SizingPolydispersity, IsothermalStability, or ColloidalStability, automatically set to Null.
TemperatureRamping
The type of temperature ramp. Linear temperature ramps increase temperature at a constant rate given by TemperatureRampRate. Step temperature ramps increase the temperature by TemperatureRampStep/TemperatureRampStepTime and holds the temperature constant for TemperatureRampStepHold before measurement.
Default Calculation: When AssayType is MeltingCurve, automatically set to Step if NumberOfTemperatureRampSteps or StepHoldTime are specified and to Linear if they are not specified. When AssayType is SizingPolydispersity, IsothermalStability, or ColloidalStability, automatically set to Null.
TemperatureRampRate
The absolute value of the rate at which the temperature is changed in the course of one heating/cooling cycle.
Default Calculation: When AssayType is MeltingCurve, if TemperatureRamping is Linear, automatically set to 1 Celsius/Minute (when AssayFormFactor is Capillary) or 0.25 Celsius/Minute (when AssayFormFactor is Plate), and if TemperatureRamping is Step, automatically set to the max ramp rate available on the instrument.
Pattern Description: Greater than or equal to 0.0015 degrees Celsius per second and less than or equal to 3.4 degrees Celsius per second or Null.
TemperatureResolution
The amount by which the temperature is changed between each data point and the subsequent data point for a given sample during the heating/cooling curves.
Default Calculation: When AssayType is MeltingCurve, automatically set to highest possible resolution available for the Instrument. When AssayType is SizingPolydispersity, IsothermalStability, or ColloidalStability, automatically set to Null.
Pattern Description: Greater than or equal to 0 degrees Celsius and less than or equal to 100 degrees Celsius or Null.
NumberOfTemperatureRampSteps
Default Calculation: When AssayType is MeltingCurve, if TemperatureRamping is Step, automatically set to the absolute value of the difference in MinTemperature and MaxTemperature divided by 1 Celsius, rounded to the nearest integer; if TemperatureRamping is Linear, automatically set to Null.
Pattern Description: Greater than or equal to 1 and less than or equal to 100 in increments of 1 or Null.
StepHoldTime
The length of time samples are held at each temperature during a stepped temperature ramp prior to light scattering measurement.
Default Calculation: When AssayType is MeltingCurve, automatically set to 30 Second when TemperatureRamping is set to Step and Null when TemperatureRamping is set to Linear.
Pattern Description: Greater than or equal to 1 second and less than or equal to 3.75 hours or Null.
Sample Storage
SampleLoadingPlateStorageCondition
The conditions under which any leftover samples from the StandardDilutionCurve or SerialDilutionCurve are stored in the SampleLoadingPlate after the samples are transferred to the AssayContainer(s).
Default Calculation: Automatically set to Null when AssayFormFactor is Plate and to Disposal when AssayFormFactor is Capillary.
Pattern Description: {{AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting}, Disposal} or an object of type or subtype Model[StorageCondition] or Null.
Programmatic Pattern: ((ObjectP[Model[StorageCondition]] | (SampleStorageTypeP | Disposal)) | Automatic) | Null
SamplesInStorageCondition
The non-default conditions under which the SamplesIn of this experiment should be stored after the protocol is completed.
Pattern Description: {{AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting}, Disposal} or an object of type or subtype Model[StorageCondition] or Null.
Programmatic Pattern: ((ObjectP[Model[StorageCondition]] | (SampleStorageTypeP | Disposal)) | Automatic) | Null
SamplesOutStorageCondition
The non-default conditions under which the SamplesOut of this experiment should be stored after the protocol is completed.
Default Calculation: When AssayFormFactor is Capillary, automatically set to Disposal. When AssayFormFactor is Plate, if all of the experiment samples have the same StorageCondition, automatically set to that StorageCondition; if the experiment samples have different StorageConditions, automatically set to Disposal; if no StorageConditions are available for the experiment samples, automatically set to AmbientStorage.
Pattern Description: {{AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting}, Disposal} or an object of type or subtype Model[StorageCondition] or Null.
Programmatic Pattern: ((ObjectP[Model[StorageCondition]] | (SampleStorageTypeP | Disposal)) | Automatic) | Null
Model Input
PreparedModelContainer
Indicates the container in which a Model[Sample] specified as input to the experiment function will be prepared.
Default Calculation: If PreparedModelAmount is set to All and when the input model has a product associated with both Amount and DefaultContainerModel populated, automatically set to the DefaultContainerModel value in the product. Otherwise set to Model[Container, Vessel, "2mL Tube"].
PreparedModelAmount
Indicates the amount of a Model[Sample] specified as input to the experiment function that will be prepared in the PreparedModelContainer. When set to All and the input model sample is not preparable, the entire amount of the input model sample that comes from one of the Products is prepared. The selected product must have both Amount and DefaultContainerModel populated, and it must not be a KitProduct. When set to All and the input model is preparable such as water, 1 Milliliter of the input model sample is prepared.
Sample Prep Options
Sample Preparation
PreparatoryUnitOperations
Specifies a sequence of transferring, aliquoting, consolidating, or mixing of new or existing samples before the main experiment. These prepared samples can be used in the main experiment by referencing their defined name. For more information, please reference the documentation for ExperimentSamplePreparation.
Pattern Description: List of one or more unit Operation ManualSamplePreparation or RoboticSamplePreparation or unit Operation must match SamplePreparationP entries or Null.
Programmatic Pattern: {((ManualSamplePreparationMethodP | RoboticSamplePreparationMethodP) | SamplePreparationP)..} | Null
Preparatory Incubation
Incubate
Indicates if the SamplesIn should be incubated at a fixed temperature prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Incubation options are set. Otherwise, resolves to False.
IncubationTemperature
Temperature at which the SamplesIn should be incubated for the duration of the IncubationTime prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 500 degrees Celsius or Null.
Programmatic Pattern: ((Ambient | RangeP[$MinIncubationTemperature, $MaxIncubationTemperature]) | Automatic) | Null
IncubationTime
Duration for which SamplesIn should be incubated at the IncubationTemperature, prior to starting the experiment.
Mix
Default Calculation: Automatically resolves to True if any Mix related options are set. Otherwise, resolves to False.
MixType
Default Calculation: Automatically resolves based on the container of the sample and the Mix option.
Pattern Description: Roll, Vortex, Sonicate, Pipette, Invert, Stir, Shake, Homogenize, Swirl, Disrupt, or Nutate or Null.
MixUntilDissolved
Indicates if the mix should be continued up to the MaxIncubationTime or MaxNumberOfMixes (chosen according to the mix Type), in an attempt dissolve any solute. Any mixing/incubation will occur prior to starting the experiment.
Default Calculation: Automatically resolves to True if MaxIncubationTime or MaxNumberOfMixes is set.
MaxIncubationTime
Maximum duration of time for which the samples will be mixed while incubated in an attempt to dissolve any solute, if the MixUntilDissolved option is chosen. This occurs prior to starting the experiment.
Default Calculation: Automatically resolves based on MixType, MixUntilDissolved, and the container of the given sample.
IncubationInstrument
Default Calculation: Automatically resolves based on the options Mix, Temperature, MixType and container of the sample.
Pattern Description: An object of type or subtype Model[Instrument, Roller], Model[Instrument, OverheadStirrer], Model[Instrument, Vortex], Model[Instrument, Shaker], Model[Instrument, BottleRoller], Model[Instrument, Roller], Model[Instrument, Sonicator], Model[Instrument, HeatBlock], Model[Instrument, Homogenizer], Model[Instrument, Disruptor], Model[Instrument, Nutator], Model[Instrument, Thermocycler], Model[Instrument, EnvironmentalChamber], Model[Instrument, Pipette], Object[Instrument, Roller], Object[Instrument, OverheadStirrer], Object[Instrument, Vortex], Object[Instrument, Shaker], Object[Instrument, BottleRoller], Object[Instrument, Roller], Object[Instrument, Sonicator], Object[Instrument, HeatBlock], Object[Instrument, Homogenizer], Object[Instrument, Disruptor], Object[Instrument, Nutator], Object[Instrument, Thermocycler], Object[Instrument, EnvironmentalChamber], or Object[Instrument, Pipette] or Null.
AnnealingTime
Minimum duration for which the SamplesIn should remain in the incubator allowing the system to settle to room temperature after the IncubationTime has passed but prior to starting the experiment.
IncubateAliquotContainer
The desired type of container that should be used to prepare and house the incubation samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
IncubateAliquotDestinationWell
The desired position in the corresponding IncubateAliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
IncubateAliquot
The amount of each sample that should be transferred from the SamplesIn into the IncubateAliquotContainer when performing an aliquot before incubation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Centrifugation
Centrifuge
Indicates if the SamplesIn should be centrifuged prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Centrifuge options are set. Otherwise, resolves to False.
CentrifugeInstrument
Pattern Description: An object of type or subtype Model[Instrument, Centrifuge] or Object[Instrument, Centrifuge] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, Centrifuge], Object[Instrument, Centrifuge]}] | Automatic) | Null
CentrifugeIntensity
The rotational speed or the force that will be applied to the samples by centrifugation prior to starting the experiment.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
CentrifugeTime
CentrifugeTemperature
The temperature at which the centrifuge chamber should be held while the samples are being centrifuged prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -10 degrees Celsius and less than or equal to 40 degrees Celsius or Null.
CentrifugeAliquotContainer
The desired type of container that should be used to prepare and house the centrifuge samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
CentrifugeAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
CentrifugeAliquot
The amount of each sample that should be transferred from the SamplesIn into the CentrifugeAliquotContainer when performing an aliquot before centrifugation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Filtering
Filtration
Indicates if the SamplesIn should be filter prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Filter options are set. Otherwise, resolves to False.
FiltrationType
Default Calculation: Will automatically resolve to a filtration type appropriate for the volume of sample being filtered.
FilterInstrument
Default Calculation: Will automatically resolved to an instrument appropriate for the filtration type.
Pattern Description: An object of type or subtype Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], or Object[Instrument, SyringePump] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], Object[Instrument, SyringePump]}] | Automatic) | Null
Filter
The filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Will automatically resolve to a filter appropriate for the filtration type and instrument.
Pattern Description: An object of type or subtype Model[Container, Plate, Filter], Model[Container, Vessel, Filter], or Model[Item, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Plate, Filter], Model[Container, Vessel, Filter], Model[Item, Filter]}] | Automatic) | Null
FilterMaterial
The membrane material of the filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter material for the given sample is Filtration is set to True.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
PrefilterMaterial
The material from which the prefilter filtration membrane should be made of to remove impurities from the SamplesIn prior to starting the experiment.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
FilterPoreSize
The pore size of the filter that should be used when removing impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter pore size for the given sample is Filtration is set to True.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
PrefilterPoreSize
The pore size of the filter; all particles larger than this should be removed during the filtration.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
FilterSyringe
Default Calculation: Resolves to an syringe appropriate to the volume of sample being filtered, if Filtration is set to True.
Pattern Description: An object of type or subtype Model[Container, Syringe] or Object[Container, Syringe] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container, Syringe], Object[Container, Syringe]}] | _String) | Automatic) | Null
FilterHousing
The filter housing that should be used to hold the filter membrane when filtration is performed using a standalone filter membrane.
Default Calculation: Resolve to an housing capable of holding the size of the membrane being used, if filter with Membrane FilterType is being used and Filtration is set to True.
Pattern Description: An object of type or subtype Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], or Object[Instrument, FilterBlock] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], Object[Instrument, FilterBlock]}] | Automatic) | Null
FilterIntensity
Default Calculation: Will automatically resolve to 2000 GravitationalAcceleration if FiltrationType is Centrifuge and Filtration is True.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
FilterTime
Default Calculation: Will automatically resolve to 5 Minute if FiltrationType is Centrifuge and Filtration is True.
FilterTemperature
The temperature at which the centrifuge chamber will be held while the samples are being centrifuged during filtration.
Default Calculation: Will automatically resolve to 22 Celsius if FiltrationType is Centrifuge and Filtration is True.
FilterContainerOut
The desired container filtered samples should be produced in or transferred into by the end of filtration, with indices indicating grouping of samples in the same plates, if desired.
Default Calculation: Automatically set as the PreferredContainer for the Volume of the sample. For plates, attempts to fill all wells of a single plate with the same model before using another one.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or {Index, Container} or Null.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | {GreaterEqualP[1, 1] | Automatic, (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
FilterAliquotContainer
The desired type of container that should be used to prepare and house the filter samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquot
The amount of each sample that should be transferred from the SamplesIn into the FilterAliquotContainer when performing an aliquot before filtration.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
FilterSterile
Default Calculation: Resolve to False if Filtration is indicated. If sterile filtration is desired, this option must manually be set to True.
Aliquoting
Aliquot
Indicates if aliquots should be taken from the SamplesIn and transferred into new AliquotSamples used in lieu of the SamplesIn for the experiment. Note that if NumberOfReplicates is specified this indicates that the input samples will also be aliquoted that number of times. Note that Aliquoting (if specified) occurs after any Sample Preparation (if specified).
AliquotAmount
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container if a liquid, or the current Mass or Count if a solid or counted item, respectively.
Programmatic Pattern: ((RangeP[1*Microliter, 20*Liter] | RangeP[1*Milligram, 20*Kilogram] | GreaterP[0*Unit, 1*Unit] | GreaterP[0., 1.] | All) | Automatic) | Null
TargetConcentration
The desired final concentration of analyte in the AliquotSamples after dilution of aliquots of SamplesIn with the ConcentratedBuffer and BufferDiluent which should be used in lieu of the SamplesIn for the experiment.
TargetConcentrationAnalyte
Default Calculation: Automatically set to the first value in the Analytes field of the input sample, or, if not populated, to the first analyte in the Composition field of the input sample, or if none exist, the first identity model of any kind in the Composition field.
Pattern Description: An object of type or subtype Model[Molecule], Model[Molecule, cDNA], Model[Molecule, Oligomer], Model[Molecule, Transcript], Model[Molecule, Protein], Model[Molecule, Protein, Antibody], Model[Molecule, Carbohydrate], Model[Molecule, Polymer], Model[Resin], Model[Resin, SolidPhaseSupport], Model[Lysate], Model[ProprietaryFormulation], Model[Virus], Model[Cell], Model[Cell, Mammalian], Model[Cell, Bacteria], Model[Cell, Yeast], Model[Tissue], Model[Material], or Model[Species] or Null.
AssayVolume
Default Calculation: Automatically determined based on Volume and TargetConcentration option values.
Pattern Description: Greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
ConcentratedBuffer
The concentrated buffer which should be diluted by the BufferDilutionFactor in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferDilutionFactor
The dilution factor by which the concentrated buffer should be diluted in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: If ConcentratedBuffer is specified, automatically set to the ConcentratedBufferDilutionFactor of that sample; otherwise, set to Null.
BufferDiluent
The buffer used to dilute the aliquot sample such that ConcentratedBuffer is diluted by BufferDilutionFactor in the final solution. The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AssayBuffer
The buffer that should be added to any aliquots requiring dilution, where the volume of this buffer added is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is not specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AliquotSampleStorageCondition
The non-default conditions under which any aliquot samples generated by this experiment should be stored after the protocol is completed.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
DestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
Pattern Description: Any well from A1 to H12 or list of one or more any well from A1 to H12 or any well from A1 to H12 entries or Null.
Programmatic Pattern: ((WellPositionP | {((Automatic | Null) | WellPositionP)..}) | Automatic) | Null
AliquotContainer
The desired type of container that should be used to prepare and house the aliquot samples, with indices indicating grouping of samples in the same plates, if desired. This option will resolve to be the length of the SamplesIn * NumberOfReplicates.
Default Calculation: Automatically set as the PreferredContainer for the AssayVolume of the sample. For plates, attempts to fill all wells of a single plate with the same model before aliquoting into the next.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null or {Index, Container} or list of one or more an object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null entries or list of one or more Automatic or Null or {Index, Container} entries.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null) | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | {((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null))..} | {({GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | (Automatic | Null))..}) | Automatic) | Null
AliquotPreparation
Default Calculation: Automatic resolution will occur based on manipulation volumes and container types.
ConsolidateAliquots
Protocol Options
Organizational Information
Template
A template protocol whose methodology should be reproduced in running this experiment. Option values will be inherited from the template protocol, but can be individually overridden by directly specifying values for those options to this Experiment function.
Pattern Description: An object of type or subtype Object[Protocol] or an object of type or subtype of Object[Protocol] with UnresolvedOptions, ResolvedOptions specified or Null.
Programmatic Pattern: (ObjectP[Object[Protocol]] | FieldReferenceP[Object[Protocol], {UnresolvedOptions, ResolvedOptions}]) | Null
Name
A object name which should be used to refer to the output object in lieu of an automatically generated ID number.
Post Experiment
MeasureWeight
Indicates if any solid samples that are modified in the course of the experiment should have their weights measured and updated after running the experiment. Please note that public samples are weighed regardless of the value of this option.
MeasureVolume
Indicates if any liquid samples that are modified in the course of the experiment should have their volumes measured and updated after running the experiment. Please note that public samples are volume measured regardless of the value of this option.
ImageSample
Example Results
Example Calls
Obtain z-average diameter, polydispersity, and mass intensity data from a single time point.
Setting the AssayType to SizingPolydispersity makes DLS measurements at a single time point and temperature:
Obtain z-average diameter and polydispersity data over time at a single temperature by setting AssayType to IsothermalStability.
The length of the IsothermalStability assay can be set by specifying the IsothermalMeasurements and MeasurementDelayTime options:
The length of the IsothermalStability assay can also be set by specifying the IsothermalRunTime option:
Obtain information about colloidal stability by setting the AssayType to ColloidalStability.
B22 (also known as A2) and kD (at analyte concentrations below 20 mg/mL) or G22 (at analyte concentrations above 20 mg/mL) provide a measure of weak protein-protein attraction or repulsion:
Sample Dilution
Generate a serial dilution curve to dilute the sample. Provide the volume of the sample, a dilution factor and the number of times the sample should be diluted. This should be used if you want the sample to be diluted by the same factor at each step of the serial dilution. The volume of the sample will be the final volume of the sample after the dilution is performed. The example call below will create a series of samples with dilution factors of (1.2, 1.44, 1.73, 2.07, 2.45, 2.99) and a volume of 60 Microliters.
Generate a serial dilution curve to dilute the sample. Provide the volume of the sample and a series of dilution factors the sample should be diluted by at each step. This should be used if you want the sample to be diluted by different factors at each step of the serial dilution. The volume of the sample will be the final volume of the sample after the dilution is performed. The example call below will create a series of samples with dilution factors of (1.2, 1.44, 1.73, 1.90, 2.09, 2.30) and a volume of 60 Microliters.
Generate a serial dilution curve to dilute the sample. Provide the volume of the sample that should be transferred, the volume of diluent the sample should be added to and the number of times the sample should be diluted. This should be used if you want give the volumes the sample will be diluted by with instead of dilution factors. The example call below will create a series of samples with dilution factors of (1.5, 2.25, 3.38, 5.06, 7.59, 11.40) and a volume of 50 Microliters.
Generate a dilution curve to dilute the sample. Provide the volume of the sample and target concentration at each point of the curve:
Obtain information about midpoint transition temperature, aggregation temperature, and temperature of onset of unfolding by collecting DLS and SLS measurements over a range of temperatures
Warnings and Errors
Messages (64)
ConflictingDLSAssayTypeNullOptions (2)
ConflictingDLSAssayTypeOptions (1)
ConflictingDLSAssayTypeSampleVolumeOptions (1)
ConflictingDLSDilutionCurves (1)
ConflictingDLSDilutionMixOptions (1)
ConflictingDLSDilutionMixTypeInstrument (1)
ConflictingDLSDilutionMixTypeOptions (1)
ConflictingDLSFormFactorCapillaryLoading (1)
ConflictingDLSFormFactorInstrument (1)
ConflictingDLSFormFactorLoadingPlate (1)
ConflictingDLSFormFactorPlateStorage (1)
ConflictingDLSFormFactorReplicateDilutionCurve (1)
ConflictingDLSIsothermalOptions (1)
ConflictingDLSIsothermalTimeOptions (1)
ConflictingFormFactorAssayContainers (2)
ConflictingFormFactorCalibratePlate (1)
ConflictingFormFactorWellCoverHeating (1)
ConflictingRampOptions (1)
DLSBufferAndBlankBufferDiffer (1)
DLSBufferNotSpecified (1)
DLSConflictingLaserSettings (1)
DLSDilutionCurveLoadingPlateMismatch (1)
DLSInputSampleNoAnalyte (2)
For ColloidalStability assays, each input sample must have a Model[Molecule,Protein] or Model[Molecule,Polymer] in its Composition, or have the Analyte option specified:
When CollectStaticLightScattering in a Plate format, each input sample must have a Model[Molecule,Protein] or Model[Molecule,Polymer] in its Composition, or have the Analyte option specified:
DLSInputSampleNoAnalyteMassConcentration (2)
For ColloidalStability assays, the Analyte must have an associated amount in the Composition field, or have the AnalyteMassConcentration option specified:
When CollectStaticLightScattering in a Plate format, the Analyte must have an associated amount in the Composition field, or have the AnalyteMassConcentration option specified:
DLSInputSampleNoAnalyteMolecularWeight (2)
DLSIsothermalAssayTooLong (1)
DLSMeasurementDelayTimeTooLow (1)
DLSMoreThanOneCompositionAnalyte (1)
DLSNotEnoughDilutionCurveVolume (1)
DLSOnlyOneDilutionCurveConcentration (1)
DLSTooManyDilutionWellsRequired (1)
InvalidAssayContainers (4)
Raise an error if we were given more than one plate type assay container:
Raise an error if we were given more than three capillary type assay containers:
Raise an error if we were given a mix of plate type and capillary type assay containers:
Raise an error if we were given an invalid dls assay container model:
InvalidCapillaryAssayContainerContents (1)
InvalidDLSInstrument (1)
InvalidDLSTemperature (1)
LowDLSReplicateConcentrations (1)
MinMaxTemperatureMismatch (1)
NonDefaultDLSSampleLoadingPlate (1)
NotEnoughAssayContainerSpace (1)
NoWellCover (1)
ObjectDoesNotExist (6)
Throw a message if we have a sample that does not exist (name form):
Throw a message if we have a container that does not exist (name form):
Throw a message if we have a sample that does not exist (ID form):
Throw a message if we have a container that does not exist (ID form):
Do NOT throw a message if we have a simulated sample but a simulation is specified that indicates that it is simulated:
Do NOT throw a message if we have a simulated container but a simulation is specified that indicates that it is simulated:
SolventNameMismatch (1)
SpecifiedCapillaryAssayContainers (1)
TooManyDLSInputs (2)
The NumberOfReplicates times the number of input samples cannot be larger than 48 when the AssayFormFactor is Capillary and the AssayType is SizingPolydispersity, MeltingCurve, or IsothermalStability:
The NumberOfReplicates times the number of input samples cannot be larger than 384 when the AssayFormFactor is Plate and the AssayType is SizingPolydispersity, MeltingCurve, or IsothermalStability:
Last modified on Mon 18 Aug 2025 14:54:20