ExperimentFluorescenceSpectroscopy
ExperimentFluorescenceSpectroscopy[Samples]⟹Protocol
generates a Protocol for measuring fluorescence intensity of the provided Samples over a range of excitation or emission wavelengths.
Fluorescence spectroscopy is a fluorescence-based microplate assay for measuring the excitation or emission spectra of a sample. For a emission spectra measurement, a fixed excitation wavelength illuminates the sample and a range of emission wavelengths is detected. For a excitation spectra measurement, the emission wavelength is fixed and a range of excitation wavelengths illuminate the sample.
Experimental Principles

Figure 1.1: Procedural overview of a FluorescenceSpectroscopy experiment. Step 1: Samples are prepared in a reader-compatible shallow well plate. Step 2: Injection lines are cleaned and primed. Step 3: The assay plate is optionally held at a specified temperature. Step 4: Injection samples are dispensed into the assay plate. Step 5: The assay plate is shaken. Step 6: Excitation and/or emission spectra are measured. Step 7: Optionally follow-up analysis such as peak detection can be performed.
Instrumentation

CLARIOstar


Figure 2.1.1: The CLARIOstar is primarily a monochromator-based plate reader and can thus support arbitrary combinations of excitation and emission wavelengths. Up to five wavelength pairs can be read in a single run. The CLARIOstar also has specialty excitation and emission filters which be used for particularly sensitive assays, including TR-FRET.
The plate chamber can be heated up to 45°C and it can mix the plate at up to 700 RPM before and/or during the run.
The reader has two 300μL syringe pump injectors which can be used for 0.5 - 300 μL injections of two unique samples at up to 4 time points during the run.
There are two primary kinetics read modes. It can perform all readings, injections and mixing for a single well before moving onto the next well or it can read in cycles, reading all the assay wells in the plate again and again until the RunTime has been reached. The second mode is generally recommended for assays except those that use very fast kinetics.
Experiment Options
General
AdjustmentSample
The sample which should be used to perform automatic adjustments of gain and/or focal height values at run time. The focal height will be set so that the highest signal-to-noise ratio can be achieved for the AdjustmentSample. The gain will be set such that the AdjustmentSample fluoresces at the specified percentage of the instrument's dynamic range. When multiple aliquots of the same sample is used in the experiment, an index can be specified to use the desired aliquot for adjustments. When set to FullPlate, all wells of the assay plate are scanned and the well of the highest fluorescence intensity if perform gain and focal height adjustments.
Pattern Description: An object of type or subtype Object[Sample] or a prepared sample or FullPlate or {Index, Sample} or Null.
Programmatic Pattern: ((FullPlate | (ObjectP[Object[Sample]] | _String) | {RangeP[1, 384, 1], ObjectP[Object[Sample]] | _String}) | Automatic) | Null
RetainCover
Indicates if the plate seal or lid on the assay container should not be taken off during measurement to decrease evaporation. When this option is set to True, injections cannot be performed as it's not possible to inject samples through the cover.
ReadLocation
Temperature
The temperature at which the experiment will be performed, if using a plate reader with temperature incubation controls.
Pattern Description: Ambient or greater than or equal to 25 degrees Celsius and less than or equal to 45 degrees Celsius.
EquilibrationTime
The length of time for which the assay plates should equilibrate at the assay temperature in the plate reader before being read.
Default Calculation: Defaults to 0 Second if Temperature is set to Ambient. Otherwise defaults to 5 Minute.
NumberOfReadings
Number of redundant readings which should be taken by the detector to determine a single averaged fluorescence intensity reading.
FocalHeight
The distance from the bottom of the plate carrier to the focal point. If set to Automatic, the focal height will be adjusted based on the Gain adjustment, which will occur only if the gain values are set to percentages.
Pattern Description: Auto or greater than or equal to 0 millimeters and less than or equal to 25 millimeters or Null.
WorkCell
Preparation
Indicates if this unit operation is carried out primarily robotically or manually. Manual unit operations are executed by a laboratory operator and robotic unit operations are executed by a liquid handling work cell.
NumberOfReplicates
Number of times each of the input samples should be analyzed using identical experimental parameters.
ExcitationScanGain
The gain which should be applied to the signal reaching the primary detector during the excitation scan. This may be specified either as a direct voltage, or as a percentage (which indicates that the gain should be set such that the AdjustmentSample fluoresces at that percentage of the instrument's dynamic range).
Default Calculation: If unspecified defaults to 90% if an adjustment sample is provided or if the instrument can scan the entire plate to determine gain. Otherwise defaults to 2500 Microvolt
Pattern Description: Greater than or equal to 0 microvolts and less than or equal to 4095 microvolts or greater than or equal to 0 percent and less than or equal to 95 percent or Null.
Programmatic Pattern: ((RangeP[0*Percent, 95*Percent] | RangeP[0*Microvolt, 4095*Microvolt]) | Automatic) | Null
EmissionScanGain
The gain which should be applied to the signal reaching the primary detector during the emission scan. This may be specified either as a direct voltage, or as a percentage (which indicates that the gain should be set such that the AdjustmentSample fluoresces at that percentage of the instrument's dynamic range).
Default Calculation: If unspecified defaults to 90% if an adjustment sample is provided or if the instrument can scan the entire plate to determine gain. Otherwise defaults to 2500 Microvolt
Pattern Description: Greater than or equal to 0 microvolts and less than or equal to 4095 microvolts or greater than or equal to 0 percent and less than or equal to 95 percent or Null.
Programmatic Pattern: ((RangeP[0*Percent, 95*Percent] | RangeP[0*Microvolt, 4095*Microvolt]) | Automatic) | Null
Sample Handling
MoatSize
Indicates the number of concentric perimeters of wells which should be should be filled with MoatBuffer in order to decrease evaporation from the assay samples during the run.

Figure 3.1: Use the moat options, MoatBuffer, MoatVolume and MoatSize to create an outer ring of wells filled with buffer. This has been shown to decrease evaporation during long reads.
MoatVolume
Default Calculation: Automatically set to the RecommendedFillVolume of the assay plate if informed, or 75% of the MaxVolume of the assay plate if not, if any other moat options are specified.
MoatBuffer
Pattern Description: An object of type or subtype Object[Sample] or Model[Sample] or a prepared sample or Null.
PrimaryInjectionSample
The sample to be injected in the first round of injections in order to introduce a time sensitive reagent/sample into the plate before/during fluorescence measurement.
Pattern Description: An object of type or subtype Object[Sample] or Model[Sample] or a prepared sample or Null.
SecondaryInjectionSample
Pattern Description: An object of type or subtype Object[Sample] or Model[Sample] or a prepared sample or Null.
PrimaryInjectionVolume
Pattern Description: Greater than or equal to 0.5 microliters and less than or equal to 300 microliters or Null.
SecondaryInjectionVolume
Pattern Description: Greater than or equal to 0.5 microliters and less than or equal to 300 microliters or Null.
PrimaryInjectionFlowRate
The speed at which to transfer injection samples into the assay plate in the first round of injections.
Pattern Description: 430 microliters per second, 400 microliters per second, 350 microliters per second, 300 microliters per second, 260 microliters per second, 220 microliters per second, 190 microliters per second, 170 microliters per second, 150 microliters per second, 135 microliters per second, 115 microliters per second, 100 microliters per second, 85 microliters per second, 65 microliters per second, or 50 microliters per second or Null.
SecondaryInjectionFlowRate
The speed at which to transfer injection samples into the assay plate in the second round of injections.
Pattern Description: 430 microliters per second, 400 microliters per second, 350 microliters per second, 300 microliters per second, 260 microliters per second, 220 microliters per second, 190 microliters per second, 170 microliters per second, 150 microliters per second, 135 microliters per second, 115 microliters per second, 100 microliters per second, 85 microliters per second, 65 microliters per second, or 50 microliters per second or Null.
PlateReaderMix
PlateReaderMixTime
Default Calculation: Automatically set to 30 second if any other plate reader mix options are specified.
PlateReaderMixRate
Default Calculation: Automatically set to 700 RPM if any other plate reader mix options are specified.
Pattern Description: Greater than or equal to 100 revolutions per minute and less than or equal to 700 revolutions per minute or Null.
PlateReaderMixMode
Default Calculation: Automatically set to DoubleOrbital if any other plate reader mix options are specified.
ReadDirection
Indicates the order in which wells should be read by specifying the plate path the instrument should follow when measuring fluorescence.

Figure 3.2: Use the ReadDirection option to control how the plate reader scans the plate during each read cycle. Choosing a more efficient path will lead to a shorter DetectionInterval. Also note that you may want to consider dripping injectors if you're working with low viscosity injection samples.
InjectionSampleStorageCondition
The non-default conditions under which any injection samples used by this experiment should be stored after the protocol is completed.
Pattern Description: {AmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, BacteriaIncubation, MammalianIncubation, TissueCultureCellsIncubation, MicrobialCellsIncubation, MicrobialCellsShakingIncubation, YeastCellsIncubation, YeastCellsShakingIncubation, ViralIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
Sampling
SamplingPattern
SamplingDistance
Default Calculation: Automatically resolves to Null if SamplingPattern is set to Center otherwise resolves to 80% of the diameter of the well.
Pattern Description: Greater than or equal to 1 millimeter and less than or equal to 6 millimeters or Null.
SamplingDimension
Specifies the size of the grid used for Matrix sampling. For example SamplingDimension->5 will scan a 5 x 5 grid.
Post Experiment
SamplesInStorageCondition
The non-default conditions under which the SamplesIn of this experiment should be stored after the protocol is completed. If left unset, SamplesIn will be stored according to their current StorageCondition.
Pattern Description: {AmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, BacteriaIncubation, MammalianIncubation, TissueCultureCellsIncubation, MicrobialCellsIncubation, MicrobialCellsShakingIncubation, YeastCellsIncubation, YeastCellsShakingIncubation, ViralIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
Spectral Scanning
Instrument
Pattern Description: An object of type or subtype Model[Instrument, PlateReader] or Object[Instrument, PlateReader]
AdjustmentEmissionWavelength
The wavelength at which fluorescence should be read in order to perform automatic adjustments of gain and focal height values at run time.
Default Calculation: If unspecified uses the wavelength in the middle of the requested emission wavelength range if the gain is not being set directly.
Pattern Description: Greater than or equal to 320 nanometers and less than or equal to 740 nanometers or Null.
AdjustmentExcitationWavelength
The wavelength at which the sample should be excited in order to perform automatic adjustments of gain and focal height values at run time.
Default Calculation: If unspecified uses the wavelength in the middle of the requested excitation wavelength range if the gain is not being set directly.
Pattern Description: Greater than or equal to 320 nanometers and less than or equal to 740 nanometers or Null.
Optics
SpectralScan
Indicates if fluorescence should be recorded using a fixed excitation wavelength and a range of emission wavelengths (Emission) or using a fixed emission wavelength and a range of excitation wavelengths (Excitation). Specify Emission and Excitation to record both spectra.
Default Calculation: If unspecified uses the emission spectrum if EmissionWavelengthRange is specified and/or the excitation spectrum if ExcitationWavelengthRange is specified, defaulting to measure both spectrum if all options are left Automatic.
ExcitationWavelength
The wavelength which should be used to excite fluorescence in the samples. Fluorescence will then be measured over 'EmissionWavelengthRange'.
Default Calculation: If unspecified uses the wavelength 50 Nanometer below the smallest requested emission wavelength.
Pattern Description: Greater than or equal to 320 nanometers and less than or equal to 740 nanometers or Null.
EmissionWavelength
The wavelength at which fluorescence emitted from the sample should be measured after the sample has been excited by each wavelength in 'ExcitationWavelengthRange'.
Default Calculation: If unspecified uses the wavelength 50 Nanometer above the largest requested excitation wavelength.
Pattern Description: Greater than or equal to 320 nanometers and less than or equal to 740 nanometers or Null.
ExcitationWavelengthRange
Defines the wavelengths which should be used to excite fluorescence in the samples. Fluorescence will then be measured at 'EmissionWavelength'.
Default Calculation: If unspecified uses the the largest possible range of excitation wavelengths with the constraints that the range must be within the instrument operating limits and below the emission wavelength.
Pattern Description: A span from anything greater than or equal to 320 nanometers and less than or equal to 740 nanometers to anything greater than or equal to 320 nanometers and less than or equal to 740 nanometers or Null.
Programmatic Pattern: (RangeP[320*Nanometer, 740*Nanometer] ;; RangeP[320*Nanometer, 740*Nanometer] | Automatic) | Null
EmissionWavelengthRange
Defines the wavelengths at which fluorescence emitted from the sample should be measured after the sample has been excited by 'ExcitationWavelength'.
Default Calculation: If unspecified uses the the largest possible range of emission wavelengths with the constraints that the range must be within the instrument operating limits and above the excitation wavelength.
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 ExperimentSampleManipulation.
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
PreparatoryPrimitives
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 ExperimentSampleManipulation.
Pattern Description: List of one or more a primitive with head Define, Transfer, Mix, Aliquot, Consolidation, FillToVolume, Incubate, Filter, Wait, Centrifuge, or Resuspend entries or 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, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, Silica, or HLB 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, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, Silica, or HLB 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, 0.1 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, 0.1 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 ConcentrationFactor 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, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, BacteriaIncubation, MammalianIncubation, TissueCultureCellsIncubation, MicrobialCellsIncubation, MicrobialCellsShakingIncubation, YeastCellsIncubation, YeastCellsShakingIncubation, ViralIncubation, 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 or Null.
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 Calls
Excitation and Emission Wavelengths

Select a single emission wavelength to capture as the sample is illuminated with a spectrum of excitation wavelengths:

Injections
To specify injections, use the InjectionSample, InjectionVolume, and InjectionTime options. The sample and volume options are index matched to the input samples allowing you to control the wells which receive injections
and the volumes of those injections. In the example below, all the input samples will receive the first injection after 20 minutes.
At the 40 minute mark the first sample will receive a second injection of 75uL, the second sample will receive 25uL, and the third sample will not receive a second injection:

Plate Reader Mixing

Gain Setting

Alternatively, directly specify the voltage to apply to the PMT. This is generally more helpful in repeat experiments. The gain can be taken from Object[Data, FluorescenceSpectroscopy][Gains]

Warnings and Errors
Messages (46)
AdjustmentEmissionWavelengthOutOfRange (1)
AdjustmentEmissionWavelengthRequired (1)
AdjustmentEmissionWavelengthUnneeded (1)
AdjustmentExcitationWavelengthOutOfRange (1)
AdjustmentExcitationWavelengthRequired (1)
AdjustmentExcitationWavelengthUnneeded (1)
AdjustmentSampleIndex (1)
AdjustmentSampleUnneeded (1)
AmbiguousAdjustmentSample (1)
BottomReadingAliquotContainer (1)
CoveredTopRead (1)
FullPlateUnsupported (2)
InjectionFlowRateRequired (1)
InjectionFlowRateUnneeded (1)
InstrumentPrecision (1)
MissingInjectionInformation (2)
MixingParametersConflict (1)
MixingParametersRequired (1)
MixingParametersUnneeded (1)
MoatAliquotsRequired (1)
PlateReaderStowaways (2)
RepeatedPlateReaderSamples (1)
ReplicateAliquotsRequired (1)
SamplingCombinationUnavailable (1)
SharedContainerStorageCondition (1)
SinglePlateRequired (3)
TooManyMoatWells (1)
TooManyPlateReaderSamples (1)
Possible Issues
Evaporation
During long runs evaporation can cause signal drift over time. Additionally using a high NumberOfReadings can lead to an initial hump while the PMT heats up.

Slow Injections
Variable Detection Interval
The detection interval is determined by the instrument at run time. It depends on the number of wells being read, mixing parameters, the number of multichromatics, and the NumberOfReadings.

Last modified on Mon 19 Dec 2022 10:51:40