ExperimentChangeMedia
ExperimentChangeMedia[Samples]⟹Protocol
creates a Protocol object to change the media of the `Samples` containing living cells in order to replenish nutrients required for the cell growth.
Changing Media is the process of using a solution (usually media or buffer) to remove impurities or unwanted metabolites from a suspension or adherent cell samples. Removing unwanted metabolites is important during cell culture because they can be harmful and pontentially kill cells if left accumulating to high levels. For suspension cells, centrifugation is used to separate the cells from the previous media and metabolites. For adherent cultures, the previous media and metabolites are directly aspirated from the cell samples adherent to the surface of the container. After removing the previous media and metabolites, the cells can be washed with a buffer or media which is added to the cells, mixed, and then aspirated off again in order to remove the trace amounts of media and metabolites. The washing procedure can be repeated multiple times; after which cells can be optionally resuspended in media for further growth.
Experimental Principles
Figure 1.1: Procedural overview of a ChangeMedia experiment in which cell isolation technique is Pellet. Step 1: Source media is removed from the cell culture by centrifuging. Step 2: Wash solution is added and mixed with the cells by shaking or pipetting. Step 3: Wash solution is removed by centrifuging. Step 4: Optional media replenishment is applied after Step 2 and Step 3 being repeated for multiple times. The cell sample is optionally mixed with the fresh media by shaking or pipetting.
Figure 1.2: Procedural overview of a ChangeMedia experiment in which cell isolation technique is Aspirate. Step 1: Source media is removed from the cell culture by directly aspirating from the cell sample adherent to the surface of the container. Step 2: Wash solution is added and mixed with the cells by shaking or pipetting. Step 3: Wash solution is removed by aspirating from the cell sample. Step 4: Optional media replenishement is applied after Step 2 and Step 3 being repeated for multiple times. The cell sample is optionally mixed with the fresh media by shaking or pipetting.
Instrumentation
bioSTAR
microbioSTAR
Inheco Incubator Shaker DWP
Figure 2.3: An overview of the Inheco Incubator Shaker DWP. It is capabale of heating and mixing the contents of a single SBS plate by shaking in an enclosed space, which more ideal for warming deep well plates than heaters which heat from the bottom of the plate.
Inheco Thermoshake
Figure 2.4: An overview of the Inheco Thermoshake. It is capabale of heating or cooling while mixing the contents of a single SBS plate by shaking.
HiG4
Figure 2.5: The HiG4 robotic centrifuge has a swinging bucket rotor that can hold two SBS standard plates. Plates can be loaded and spins can be performed fully robotically. This centrifuge is accessible only using RoboticCellPreparation protocols.
Hamilton Heater Cooler
Experiment Options
General
CellType
The taxon of the organism or cell line from which the cell sample originates. Options include Bacterial, Mammalian, Insect, Plant, and Yeast.
Default Calculation: Automatically set to the CellType specified in Object[Sample] for the input cell sample.
CultureAdhesion
Indicates how the cell sample physically interacts with its container. Options include Adherent and Suspension (including any microbial liquid media).
CellIsolationTechnique
The technique used to remove impurities, debris, and media from cell samples. Suspension cells are centrifuged to separate the cells from the media or buffer. Adherent cells remain attached to the bottom of the culture plate whereas the media can be removed via aspiration.
Default Calculation: If CultureAdhesion is Adherent, then set to Aspirate. If CultureAdhesion is set to Suspension and any of the Pellet Options not Null, automatically set to Pellet.
CellIsolationInstrument
The instrument used to isolate the cell sample. Centrifuging separates cells from media, forming a cell pellet. The supernatant is removed or harvested, leaving the cell pellet in the container.
Default Calculation: Automatically set to robotic integrated centrifuge, Model[Instrument, Centrifuge, "HiG4"] if CellIsolationTechnique is set to Pellet.
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
RoboticInstrument
Default Calculation: Automatically set to Model[Instrument, LiquidHandler, "bioSTAR"] if CellType is Mammalian. Otherwise, set to Model[Instrument, LiquidHandler, "microbioSTAR"].
Pattern Description: An object of type or subtype Model[Instrument, LiquidHandler] or Object[Instrument, LiquidHandler]
Programmatic Pattern: ObjectP[{Model[Instrument, LiquidHandler], Object[Instrument, LiquidHandler]}] | Automatic
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.
WorkCell
The automated workstation with a collection of integrated instruments on which this unit operation will be will be performed if Preparation -> Robotic.
Method
Default Calculation: Automatically set to the object specified in Model[Cell][WashMethod], if any. Automatically set to Custom if no method object is specified for Model[Cell].
Media Aspiration
CellAspirationVolume
Indicates how much media to remove from an input sample of cells when isolating with Aspriate or Pellet.
Default Calculation: Automatically set to aspirate off the majority of the sample volume, leaving the smaller of 5% of the container's capacity or 100 Microliter.
Pattern Description: All or greater than or equal to 0 microliters and less than or equal to 200 milliliters.
CellIsolationTime
The amount of time to centrifuge the cell samples. Centrifuging is intended to separate cells from media.
CellPelletContainer
Default Calculation: Automatically set to the deep-well plate whose WorkingVolume is larger than the sample volume if CellIsolationTechnique is set to Pellet.
Pattern Description: Container or Container with Index or Container with Well or Container with Well and Index or Null.
Programmatic Pattern: (((ObjectP[{Object[Container], Model[Container]}] | _String) | {GreaterEqualP[1, 1], ObjectP[{Object[Container], Model[Container]}]} | {Alternatives @@ Flatten[AllWells[NumberOfWells -> $MaxNumberOfWells]], ObjectP[{Object[Container], Model[Container]}]} | {Alternatives @@ Flatten[AllWells[NumberOfWells -> $MaxNumberOfWells]], {GreaterEqualP[1, 1], ObjectP[{Model[Container]}]}}) | Automatic) | Null
CellPelletIntensity
The rotational speed or force applied to the cell sample by centrifugation. Centrifuging is intended to separate cells from media, forming a cell pellet.
Default Calculation: If CellIsolationTechnique is set to Pellet, automatically set to 300 standard accelerations due to gravity on the surface of the earth if CellType is Mammalian, 2000 standard accelerations due to gravity on the surface of the earth if CellType is Bacterial, 1000 standard accelerations due to gravity on the surface of the earth if CellType is Yeast.
Pattern Description: Greater than or equal to 0 revolutions per minute and less than or equal to 5703 revolutions per minute or greater than or equal to 0 standard accelerations due to gravity on the surface of the earth and less than or equal to 3600 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((RangeP[0*RPM, $MaxRoboticCentrifugeSpeed] | RangeP[0*GravitationalAcceleration, $MaxRoboticRelativeCentrifugalForce]) | Automatic) | Null
CellAspirationAngle
Indicates the tilting angle of the adherent cell culture plate when aspirating off the input sample media. See figure 2 of the documentation page.
Pattern Description: Greater than or equal to 0 degrees and less than or equal to 10 degrees or Null.
AliquotSourceMedia
Default Calculation: Automatically set to True if any source media options (AliquotMediaVolume, AliquotMediaContainer,...) are not Null.
AliquotMediaVolume
Default Calculation: Automatically set to the same as CellAspirationVolume if AliquotSourceMedia is True.
Pattern Description: All or greater than or equal to 0 microliters and less than or equal to 200 milliliters or Null.
AliquotMediaContainer
Default Calculation: Automatically set to the same container as the input sample if AliquotSourceMedia is True.
Pattern Description: Container or Container with Index or Container with Well or Container with Well and Index or Null.
Programmatic Pattern: (((ObjectP[{Object[Container], Model[Container]}] | _String) | {GreaterEqualP[1, 1], ObjectP[{Object[Container], Model[Container]}]} | {Alternatives @@ Flatten[AllWells[NumberOfWells -> $MaxNumberOfWells]], ObjectP[{Object[Container], Model[Container]}]} | {Alternatives @@ Flatten[AllWells[NumberOfWells -> $MaxNumberOfWells]], {GreaterEqualP[1, 1], ObjectP[{Model[Container]}]}}) | Automatic) | Null
AliquotMediaContainerLabel
A user defined word or phrase used to identify the SourceMediaContainer that contains the source media separated from cell samples, for use in future analysis.
Default Calculation: Automatically set to "source media from cell sample container #" if AliquotSourceMedia is True.
AliquotMediaStorageCondition
Programmatic Pattern: (((SampleStorageTypeP | Disposal) | ObjectP[{Model[StorageCondition]}]) | Automatic) | Null
AliquotMediaLabel
A user defined word or phrase used to identify the saved cells sample that is being isolated from media, for use in downstream unit operations.
Default Calculation: Automatically set to "source media from cell sample #" if AliquotSourceMedia is True.
Washing
NumberOfWashes
The number of times the sample is washed with WashSolution, prior to replenishment with fresh media, in order to wash trace amounts of media and metabolites from the cells.
Default Calculation: Automatically set to 3 if CultureAdhesion is set to Adherent. Otherwise, set to 2.
WashSolution
Default Calculation: Automatically set to Model[Sample,"1x PBS (Phosphate Buffer Saline), Autoclaved"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
WashSolutionTemperature
Programmatic Pattern: ((RangeP[$MinRoboticIncubationTemperature, $MaxRoboticIncubationTemperature] | Ambient) | Automatic) | Null
WashSolutionEquilibrationTime
The WashSolution will be incubated for a minimum duration of this time at WashSolutionTemperature prior to washing the live cell sample.
WashVolume
Pattern Description: Greater than or equal to 0 milliliters and less than or equal to 200 milliliters or Null.
WashMixType
Default Calculation: Automatically set to Shake if any of shake options (WashMixRate, WashMixTime, WashMixInstrument) are set. Otherwise, set to Pipette if any of the pipette options (NumberOfWashMix, WashMixVolume) are set.
WashMixInstrument
Default Calculation: Automatically set to Model[Instrument, Shaker,"Inheco ThermoshakeAC"] if the WashMixType is set to Shake.
Pattern Description: An object of type or subtype Model[Instrument, Shaker] or Object[Instrument, Shaker] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, Shaker], Object[Instrument, Shaker]}] | Automatic) | Null
WashMixTime
WashMixRate
The rate at which the sample is mixed with WashSolution via the selected WashMixType for the duration specified by WashMixTime.
Pattern Description: Greater than or equal to 100 revolutions per minute and less than or equal to 2000 revolutions per minute or Null.
WashMixVolume
The volume of the cell sample in WashSolution that is pipetted up and down to mix the live cell samples with WashSolution.
Default Calculation: If WashMixType is set to Pipette, automatically set to the smaller of half of the volume of the input sample or 970 Microliter, which is the largest volume that can be pipetted in a single step by the robotic liquid handler.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 970 microliters or Null.
NumberOfWashMixes
The number of pipetting cycles (drawing liquid up into the pipette and dispensing back down out of the pipette) used to mix the live cell samples with WashSolution.
Default Calculation: Automatically set to be the lesser of 50 or 5 * WashVolume/WashMixVolume if WashMixType is set to Pipette.
Pattern Description: Greater than or equal to 1 and less than or equal to 100 in increments of 1 or Null.
WashTemperature
Programmatic Pattern: ((RangeP[$MinRoboticIncubationTemperature, $MaxRoboticIncubationTemperature] | Ambient) | Automatic) | Null
WashAspirationVolume
Indicates how much wash solution to remove from an input sample of cells prior to media replenishment when isolating with Aspriate or Pellet.
Pattern Description: All or greater than or equal to 0 microliters and less than or equal to 200 milliliters or Null.
WashIsolationTime
The amount of time to pellet the cell samples prior to changing media. Centrifuging is intended to separate cells from media.
Default Calculation: Automatically set the same as CellIsolationTime selection if CellIsolationTechnique is set to Pellet.
WashPelletIntensity
The rotational speed or force applied to the cell sample by centrifugation prior to changing media. Centrifuging is intended to separate cells from media, forming a cell pellet.
Default Calculation: Automatically set to the same as CellPelletIntensity selection if CellIsolationTechnique is Pellet.
Programmatic Pattern: ((RangeP[0*RPM, $MaxRoboticCentrifugeSpeed] | RangeP[0*GravitationalAcceleration, $MaxRoboticRelativeCentrifugalForce]) | Automatic) | Null
WashAspirationAngle
Indicates the tilting angle of the adherent cell culture plate to aspirate off the wash buffer. The tilt causes the liquid to pool on one edge of the container, thereby making it easier to aspirate off the liquid.
Default Calculation: Automatically set to the CellAspirationAngle selection if CellIsolationTechnique is Aspirate.
Pattern Description: Greater than or equal to 0 degrees and less than or equal to 10 degrees or Null.
Media Replenishment
ResuspensionMedia
Default Calculation: Automatically set to the PreferredLiquidMedia of the cell model. Otherwise, set to the same media as the input sample object.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or None.
ResuspensionMediaTemperature
Programmatic Pattern: ((RangeP[$MinRoboticIncubationTemperature, $MaxRoboticIncubationTemperature] | Ambient) | Automatic) | Null
ResuspensionMediaEquilibrationTime
Indicates the ResuspensionMedia is incubated at ResuspensionMediaTemperature for at least this amount of time prior to resuspending the cell sample.
ResuspensionMediaVolume
Default Calculation: Automatically set to WorkingVolume of the container object. Otherwise set to 80% of the container's capacity.
Pattern Description: Greater than or equal to 0 milliliters and less than or equal to 200 milliliters or Null.
ResuspensionMixType
Indicates the style of motion (Shake or Pipette) used to resuspend the cell sample with ResuspensionMedia. If ResuspensionMixType is set to None, the cells will not be mixed with the ResuspensionMedia.
Default Calculation: Automatically set to Shake if any of ResuspensionMixRate, ResuspensionMixTime, ResuspensionMixInstrument are set. Otherwise, set to Pipette.
ResuspensionTemperature
Programmatic Pattern: ((RangeP[$MinRoboticIncubationTemperature, $MaxRoboticIncubationTemperature] | Ambient) | Automatic) | Null
ResuspensionMixInstrument
Default Calculation: Automatically set to Model[Instrument, Shaker,"Inheco ThermoshakeAC"] if ResuspensionMixType is Shake.
Pattern Description: An object of type or subtype Model[Instrument, Shaker] or Object[Instrument, Shaker] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, Shaker], Object[Instrument, Shaker]}] | Automatic) | Null
ResuspensionMixTime
ResuspensionMixRate
Default Calculation: Automatically set to 200 RPM if ResuspensionMixType is set to Shake. Automatically set to 20 RPM if ResuspensionMixType is set to 20 RPM.
Pattern Description: Greater than or equal to 100 revolutions per minute and less than or equal to 2000 revolutions per minute or Null.
ResuspensionMixVolume
The volume of the sample that is pipetted up and down to resuspend the cell sample in ResuspensionMedia.
Default Calculation: If ResuspensionMixType is set to Pipette, automatically set to the smaller of half of the volume of the input sample or 970 Microliter, which is the largest volume that can be pipetted in a single step by the robotic liquid handler.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 970 microliters or Null.
NumberOfResuspensionMixes
The number of pipetting cycles (drawing liquid up into the pipette and dispensing back down out of the pipette) used to resuspend the cell sample in ResuspensionMedia.
Default Calculation: Automatically set to be the lesser of 50 or 5 * ResuspensionMediaVolume/ResuspensionMixVolume if ResuspensionMixType is set to Pipette.
Pattern Description: Greater than or equal to 1 and less than or equal to 100 in increments of 1 or Null.
ReplateCells
ContainerOut
The container into which the resuspended cell sample is transferred into after cell washing and media replenishment.
Default Calculation: Automatically set to the same plate as the input sample if ReplateCells is True.
Pattern Description: Container or Container with Index or Container with Well or Container with Well and Index or Null.
Programmatic Pattern: (((ObjectP[{Object[Container], Model[Container]}] | _String) | {GreaterEqualP[1, 1], ObjectP[{Object[Container], Model[Container]}]} | {Alternatives @@ Flatten[AllWells[NumberOfWells -> $MaxNumberOfWells]], ObjectP[{Object[Container], Model[Container]}]} | {Alternatives @@ Flatten[AllWells[NumberOfWells -> $MaxNumberOfWells]], {GreaterEqualP[1, 1], ObjectP[{Model[Container]}]}}) | Automatic) | Null
ContainerOutLabel
A user defined word or phrase used to identify the ContainerOut that contains the replated cell samples.
Default Calculation: Automatically set to "wash cell sample container #" if ContainerOut is not Null.
SampleOutLabel
A user defined word or phrase used to identify the SampleOut that contains the cell samples after WashCells or ChangeMedia.
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, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
SamplesOutStorageCondition
The non-default conditions under which any new samples generated by this experiment should be stored after the protocol is completed. If left unset, the new samples will be stored according to their Models' DefaultStorageCondition.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
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
Methods
CultureAdhesion
CellIsolationTechnique
Pellet Options
Aspirate Options
Clarify the options for aspirating the source media from the cell sample adherent to the surface of the container:
Clarify the options for aspirating the wash solution from the cell sample adherent to the surface of the container:
NumberOfWashes
Mixing Options
Warnings and Errors
Messages (15)
AspirationVolumeConflictingOption (1)
CellIsolationTechniqueConflictingOptions (1)
ExtraneousMediaReplenishmentOptions (1)
ExtraneousWashingOptions (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:
ResuspensionMixTypeConflictingOptions (1)
TotalVolumeConflictingOption (1)
WashIsolationTechniqueConflictingOptions (1)
Last modified on Wed 8 Oct 2025 12:53:14