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"Applications", "NameofStructureSystem": "Applications", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "OWP_Techniques", "NameofStructureSystem": "Techniques", "StructureNodeID": "999-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Disease_Areas", "NameofStructureSystem": "Disease Areas", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Pathogens", "NameofStructureSystem": "Pathogens", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Health_Topics", "NameofStructureSystem": "Health Topics", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Product_Grouping", "NameofStructureSystem": "Product Grouping", "StructureNodeID": "02-0008", "StructureGroupPath": "CustomBiotech->Biologics", "StructureGroupName": "Biologics" } ] }, { "ProductID": "3.8.9.9.1.7", "BrandName": "Cedex HiRes Control Beads", "ProductNameAddition": "1×106/mL Control Beads for the Cedex HiRes Analyzer", "ReferenceType": "Reagent", "Classification": [ { "IdentifierofStructureSystem": "Product_Grouping", "NameofStructureSystem": "Product Grouping", "StructureNodeID": "02-0007", "StructureGroupPath": "CustomBiotech->Cedex HiRes Analyzer Products", "StructureGroupName": "Cedex HiRes Analyzer Products" }, { "IdentifierofStructureSystem": "OWP_Product_Types", "NameofStructureSystem": "Product Types", "StructureNodeID": "90-000-00", "StructureGroupPath": "Controls", "StructureGroupName": "Controls" }, { "IdentifierofStructureSystem": "OWP_Organization", "NameofStructureSystem": "Organization", "StructureNodeID": "01-01-01-07-00", "StructureGroupPath": "Roche Diagnostics Solution (RDS)->Customer Areas->Core lab->CustomBiotech and LifeScience", "StructureGroupName": "CustomBiotech and LifeScience" }, { "IdentifierofStructureSystem": "OWP_Family", "NameofStructureSystem": "Product Families", "StructureNodeID": "490", "StructureGroupPath": "Cedex", "StructureGroupName": "Cedex" }, { "IdentifierofStructureSystem": "Product_Solutions", "NameofStructureSystem": "Product Solutions", "StructureNodeID": "520", "StructureGroupPath": "CustomBiotech", "StructureGroupName": "CustomBiotech" }, { "IdentifierofStructureSystem": "Lab_Type", "NameofStructureSystem": "Lab Types", "StructureNodeID": "180-00", "StructureGroupPath": "Manufacturing", "StructureGroupName": "Manufacturing" }, { "IdentifierofStructureSystem": "Applications", "NameofStructureSystem": "Applications", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "OWP_Techniques", "NameofStructureSystem": "Techniques", "StructureNodeID": "999-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Disease_Areas", "NameofStructureSystem": "Disease Areas", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Pathogens", "NameofStructureSystem": "Pathogens", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Health_Topics", "NameofStructureSystem": "Health Topics", "StructureNodeID": "99-00-00", "StructureGroupPath": "NOT APPLICABLE", "StructureGroupName": "NOT APPLICABLE" }, { "IdentifierofStructureSystem": "Product_Grouping", "NameofStructureSystem": "Product Grouping", "StructureNodeID": "02-0008", "StructureGroupPath": "CustomBiotech->Biologics", "StructureGroupName": "Biologics" } ] } ] }, "ProductSpec": [ { "ProductSpecVariant": { "Chapters": [ { "Language": "en", "Value": "2.5 - 4 minutes (depending on settings).", "Name": "Assay Time" }, { "Language": "en", "Value": "
  1. Cedex HiRes Analyzer
  2. Desktop Computer with Flat Screen
  3. Latest valid Cedex Software
  4. Operator's Guide
  5. Quick Start Guide
", "Name": "Content" }, { "Language": "en", "Value": "Use the Cedex HiRes Analyzer for fast and reliable, image-based, high resolution cell culture analysis. The instrument delivers precise results for cell concentration, viability, and morphological characteristics.", "Name": "Applications" }, { "Language": "en", "Value": "Cleaning recommendations

Under normal use and conditions, Roche recommends the following routines:

  • Chamber Clean 2: Run at least 2x per day, once in the middle of the day and once at the end of the day
  • Standard Clean: Run at least 1x per day
  • LM Shutdown: Run before leaving machine to stand overnight

Less frequent routines that should be carried out include:

  • Intensive Clean: Run 1x per week
  • Intensive Clean with HCl: Can be carried out 1-2x per month, if desired

Performing Intensive Cleans

The flow chamber is made of quartz, which is inert to acids. The tubing is also inert to acids. Therefore, performing an Intensive Clean with HCl will not etch the material of the flow chamber. 0.1 to 1N HCL and 0.1 to 1N NaOH can be used. It is important avoid any drying out when using NaOH, as higher concentrations of NaOH can damage the flow chamber. Nitric acid is not recommended for use in the Cedex HiRes System.

Performing a Special Intensive Clean with HCl

When performing the special Intensive Clean with 0.1 to 1 N HCl, as described in the Cedex HiRes Analyzer Operator's Manual, Roche recommends the following: ensure that the liquid level of the Cedex Detergent in the Detergent container is at least as high as the liquid level of the HCl used for the special Intensive Clean. If the liquid level of the Cedex Detergent in the container is lower than the liquid level of the HCl, some rust may form on the outside of the metal capillary tube.

Chamber Clean 1 and Chamber Clean 2

Chamber Clean 1 is a very short cleaning routine that runs water through the chamber at a very high pressure. Chamber Clean 2 is a longer cleaning routine that flushes the chamber with water and Cedex Detergent. The Chamber Clean 1 should be tried as a quick first step if there is a possible blockage in the chamber. If Chamber Clean 1 does not clear the blockage, then Chamber Clean 2 can be used.

Fast Clean and Standard Clean

The Fast Clean cycle cleans the sample cup with Cedex Detergent and water, and rinses the flow chamber with water. Standard Clean flushes the entire system, including all of the capillaries and the flow chamber, with Cedex Cleaning Solution, Cedex Detergent and water. Fast Clean is considerably faster and can be used to quickly clean the sample cup and flow chamber before the next sample is run.

Trypan Blue storage temperature

An unopened bottle of 0.2% Trypan Blue has shelf life of up to 1 year from the production date, when stored at room temperature. The expiration date is listed on the outside package. Once the bottle is opened, the contents are useable for one week at room temperature. This means that the opened bottle can be left on a Cedex HiRes System for up to one week. Storage of opened bottles at +4 to +8°C does not change the one week shelf life limit, so there is no need for storage at +4 to +8°C after opening.

Trypan Blue concentration required

Use only 0.2% Trypan Blue solutions for Cedex HiRes Systems.

Software 2.x: Scheduling cleaning routines and possible conflict with Multi Run

Cleaning routines automatically have the lowest priority. They are always carried out after all scheduled measurements. In the case of the Multi Run, scheduled cleaning routines are performed after the Multi Run, even when a cleaning routine has been scheduled.

Software 2.5: Target drive when scheduling automated database backup
Routine database backup can be scheduled so that it is automatically included in the process queue.
Roche recommends using the D drive for scheduled database backups.

Software 2.x: Inactivity of the Scheduler

Inactivity refers to the hardware of the Cedex HiRes Analyzer. Performing cleaning routines, measurements and scans of the flow cell is considered activity. Viewing diagrams and measurements are considered as inactivity by the scheduler.

Using a new lot for retesting with Density Reference Standard Beads

Please note the following when checking the calibration of the instrument using Density Reference Standard Beads following the protocol described in the Instructions for Use: If the calculated mean Total Cell Density for the first bottle of DRSB is outside the specified range, the second bottle used to recheck the calibration must come from a different lot number. The Batch type (A or B) can be the same as the one used for the first check, as long as the actual lot number, noted on the bottle label, is different.

Archive/Restore, Data Exchange, Database backup

The Archive/Restore function can only be used to archive data from a unique database and restore it to that unique database. The function that allows exporting of a complete measurement from one database to a different database is called the \"Data Exchange\" (Export/Import) function. Both functions are accessed using the Functions menu on the main Cedex Control Center. Neither function should be used as the only way to back up measurement data. The entire database should be backed up at regular intervals using standard backup strategies. This is critical because when measurements are archived and something goes wrong with the hard disk it may not be possible to restore measurements affected (e.g., if the entire database must be replaced with a new unique database). There are two things that can be done with measurements:

\"Data Exchange\": The Export/Import options under \"Data Exchange\" can be used to export an entire measurement package from a unique database in such a way that it can then be imported into a different database. Note that measurements exported using this function cannot be imported back into the same database from which they were originally exported. They can only be imported to a different database.

\"Archive\": The Archive/Restore functions under \"Archive\" can be used to archive measurements out of a unique database to allow for deletion of measurements from the local Control Unit database. This function should be used to clear the local database of measurements to free up space on the hard drive. Note that this is not a backup strategy, because measurements can only be archived from a unique database and restored back to that same unique database. If, for example, something should happen to the computer (hard drive failure) resulting in a problem with that unique database, it may not be possible to restore measurements to the database from which they were archived. It is therefore critical to ensure that the entire database is backed up, so that archived measurements can still be restored.

Analyzing aggregates using the Aggregate Histogram

There are different ways to view data in the Aggregate Histogram. Choosing the detailed and Percent buttons shows the percentage of cells found in various aggregate sizes. The green column shows the percent live cells of a particular size (size is the number of cells per aggregate), and the red column shows the percent dead cells of a particular aggregate size. Note that the percentage live cells is the percentage of cells found in a particular aggregate size compared to the total number of live cells. When, for example, 20% is shown for live cells found in a 2-cell aggregate, this means that 20% of all live cells counted were found in 2-cell aggregates. Dead cells are not included in the total cell count for the calculation of percentage. This is also the case for dead cell aggregates: the percentage count is based on the total number of dead cells. Clicking on the absolute button displays the total number of cells (live/dead) found in each size (i.e., number of cells per aggregate). The total button combines the live and dead cells into a single histogram for each size. The percentage calculation indicates the total number of cells counted (live and dead).

Exporting diameter data

The calculated average diameter is exportable via the export to file function in the Measurement List window in software versions 2.x. Histograms showing the distribution are not exportable. It is possible to save diagrams in a variety of formats by right clicking directly in the diagram and choosing the desired format and location.

For Software versions 2.x, it is also possible to export the individual diameter data determined for each detected key in a measurement using Software versions 2.x. This is done using the Export to File function in the Measurement List window. The option Cell Data in the Export window allows for the export of information about individual keys, including key type, location, compactness and diameter.

Scheduling automated cleaning routines under a specific username

When an automated cleaning routine is scheduled via the \"Scheduling\" function in Cedex Software versions 2.x under a specific user (e.g., SuperUser), the routine is automatically added to the scheduled list for all users. It will also appear in the list of scheduled cleaning routines (the \"Schedule\" window), regardless of who is logged in or which user scheduled the routine. For example, if a cleaning routine is scheduled using the SuperUser account, the routine will be performed as scheduled, even if the user is logged into the software using a different username.", "Name": "Protocols - Help Corner" }, { "Language": "en", "Value": "Detectable cell density range: 5 x104 - 1x107 cells per ml
Detectable cell diameter range: 2 μm - 40 μm
Detectable key diameter range: 1 μm - 90 μm
Required sample volume: 300 μl
Number of samples: 1- 20", "Name": "Product Characteristics" }, { "Language": "en", "Value": "Reagent levels
\n
\nReagent levels that are too low can affect system performance. Trypan Blue must be available for accurate measurements. Cleaning routines require sufficient quantities of water, Cedex Cleaning Solution and Cedex Detergent to carry out proper cleaning routines. When cleaning reagents are too low, the flow chamber can not be properly cleaned after a measurement, resulting in inaccurate measurements later on.
\n
\nTrypan Blue effect on cell viability
\n
\nRoche tests with a variety of cell lines showed that extended exposure to Trypan Blue had little effect on cell viability, even after exposure of 100 minutes. Some cell lines may however be more sensitive to the effects of trypan blue than others. Each cell line should be tested with regard to trypan blue sensitivity.
\n
\nSample handling methods and effect on precision and accuracy
\n \n
    \n\t
  • Mixing of the cell suspension before removal of the sample for measurement
  • \n
\n
\nCells sediment quickly. Cell suspensions should be thoroughly mixed just before sample removal to avoid sampling of an nonhomogeneous solution. When multiple samples are removed from the same source, the cell suspension may need to be mixed thoroughly before the removal of each sample.
\n \n
    \n\t
  • Methods used for mixing and resuspension of the sample
  • \n
\n
\nThis is a critical step due to the risk of cell damage when cell mixing and resuspension is too vigorous. Some resuspend cells using a pipette in to break up possible aggregates, but this may lower viability. Others resuspend more gently. When different approaches are used for the same cell line, measurement results may be different. Ideally the method used for resuspending cells should be consistent.
\n \n
    \n\t
  • Removal of a sample from a bioreactor
  • \n
\n
\nDepending on the method used for removing samples from the bioreactor, there may be higher variations from sample to sample, even within a short period of time. This will depend on how effective the method is for removing a representative sample.
\n \n
    \n\t
  • Equipment used for sample handling
  • \n
\n
\nPipettes should be routinely checked and calibrated. Variation due to equipment is minimized when everyone uses the same equipment (e.g., the same pipette tips, pipette types, mixing tools etc.).
\n \n
    \n\t
  • Aggregates
  • \n
\n
\nCell cultures with many aggregates or a wide-ranging in cell diameter may not be as easy to maintain in a homogeneous suspension as cell cultures with a more uniform diameter distribution and fewer aggregates. A less homogeneous cell suspension will result in higher variation.
\n
\nImproving relative standard deviation between replicate samples
\n
\nRoche testing with cell samples containing few small aggregates achieve a %CV of approx. 5% for 5-10 replicate measurements. It may be difficult to achieve complete homogeneity for some types of cell cultures. Cell cultures with many aggregates or a large distribution in cell diameter may not be as easy to maintain in a homogeneous suspension during sample removal as cell cultures with a more uniform diameter distribution and few aggregates. A less homogeneous cell suspension, or one with many large aggregates, will result in higher replicate measurement variation.
\n
\nIt may be necessary to evaluate SOPs to determine if they require optimization. In particular, it may be necessary to test different ways of sampling cell cultures to identify the best combination of equipment, mixing routines and sample removal methods. Running various tests to test for consistency (precision) of results can help to optimize SOPs.
\n
\nThe following is an example of a series of tests that may be helpful for determining how different sampling methods affect variation between replicates:
\n \n
    \n\t
  1. Remove 1 sample from a bioreactor or cell culture vessel and measure the sample on the Cedex HiRes System. Repeat this step 5 times, removing each sample individually.
  2. \n\t
  3. Remove 10 ml from the same bioreactor or cell culture vessel. With no additional mixing, and no mixing in between sample removal, prepare 5 samples for measurement on the Cedex HiRes System.
  4. \n\t
  5. Remove 10 ml from the same bioreactor or cell culture vessel. Mix the sample by gently inverting the tube for about 5 minutes. Remove 1 sample from the tube and measure the sample on the Cedex HiRes System. Invert the tube at least 10x, remove the next sample for measurement on the Cedex HiRes System. Continue mixing before removal of each sample then measuring until 5 samples have been measured.
  6. \n
\n
\nIf the relative standard deviation for measurements carried out in steps 1 and 2 is significantly higher than that in step 3, it is likely that either samples removed from a bioreactor or cell culture vessel are not a homogeneous suspension and may require additional mixing steps. In addition, evaluate other ways to resuspend cell samples for analysis with the Cedex HiRes System to determine the most effective sample handling routine.
\n
\nLiquid management handling of the sample
\n \n
    \n\t
  • Liquid management steps are as follows
  • \n
\n\n
    \n\t
  1. Trypan Blue is taken up by syringe and injected into the 8-way valve and flow chamber. This step ensures that there is no risk of the cell sample coming in contact with Cedex Cleaning Solution, Cedex Detergent or water, which can damage cells.
  2. \n\t
  3. This initial step creates a dead volume of Trypan Blue in the 8-way valve. When the sample is taken up by syringe, the mix of dead volume and sample is exactly measured using the syringe, and excess is placed into waste.
  4. \n\t
  5. Trypan Blue is then measured and mixed 3 times with the sample/dead volume mixture remaining in the syringe after step 2.
  6. \n\t
  7. The sample/Trypan Blue mixture is pumped into the flow chamber, digital images are taken and analyzed.
  8. \n
\n \n\n
    \n\t
  • Sample volumes (300 μl) are processed in the following way in the Cedex HiRes Analyzer
  • \n
\n
\nThe syringe takes up sample and air up to 1667 μl, then discards all but 271 μl, before adding an equal amount of Trypan Blue. Note that the sample is taken up in 2 steps. The first step takes up half the sample and a dead volume (109.8 μl) of Trypan Blue. This Trypan Blue is not part of the actual sample/Trypan Blue mixing step. It is the result of the dead volume of Trypan Blue remaining in the 8-way valve (see step 2 above).
\n
\nAfter half of the sample is taken up, remaining sample is taken up by the syringe. All but 271 μL is then discarded directly into the waste; 271 μL Trypan Blue is added and then the sample is mixed to stain the cells.
\n
\nDue to the Trypan Blue dead volume in the system before the discard step, variable sample volumes can have an effect on accuracy. This effect on accuracy for a sample volume of 300 μl is taken into account by the software when calculating the results. A 5% deviation in the expected sample volume (285 μl, 315 μl) results in -1.4%, +1.3% variation, respectively; a 10% variation in sample volume (270 μl, 330 μl) results in -2.9%, +2.5% variation, respectively. The system requires at least 270 μl sample volume.
\n
\nEffect on cell viability when preparing multi samples for the Multi Run
\n
\nViability may be affected when cells are on the benchtop or on the Multi Sampler for long periods of time. This can also depend on the cell line. Many cell lines can stored at room temperature with no effect. Other cell lines may be more sensitive to long periods outside the incubator and require more careful handling. This should be tested for each cell line.
\n
\nMaximum allowed aggregation
\n
\nCounting cells in aggregates is challenging. The Software analyzes the physical size of an aggregate and divides it up into the number of cells based on how many cells (of a given size) will correspond to the size of the aggregate. the challenge is that aggregates are 3-dimensional, but the digital images are 2-dimensional. This makes counting cells in aggregates an approximation. The more cell aggregation in a culture, the less accurate the counting may be.
\n
\nGenerally, aggregate analysis is reasonably accurate when the majority are in aggregates of 2-4 cells. Aggregates of 2-4 cells are easier to identify and less likely to produce 3-dimensional problems. When large numbers of aggregates have sizes of more than 6-8 cells, there is reason to be concerned about counting accuracy. % Aggregate Rate is not the only indicator of the amount of aggregation. To obtain a more complete picture, it is best to also monitor the Aggregate Histogram for the size distribution of aggregates.
\n
\nComparing results from two different Cedex HiRes Systems
\n
\nBefore comparing results from replicate experiments performed on different Cedex HiRes Analyzers, ensure that the systems have been recently serviced. The FlowFactors for both systems should also be checked. After these checks have been carried out, results can be compared between two systems. Worn-out parts, such as worn-out syringes or 8-way valves, can affect measurement precision and accuracy.
\n
\nEffect of prolonged incubation of cell samples in Cedex Sample Cups
\n
\nMeasurement of up to 20 samples in a row requires incubation times of up to 90 minutes at room temperature in Cedex Sample Cups. In-house Roche testing using several different cell lines (CHO-K1 and several hybridoma cell lines) show no effect of prolonged incubation times on measurement results. This finding may not apply to all cell lines. For best results, test the cell line to determine the number of samples that can be measured in a row.
\n
\nDefault sedimentation duration depends on Cedex HiRes Software version
\n
\nAfter the sample is injected into the chamber, cells are allowed to settle to the bottom of the flow chamber for a specified amount of time. This is known as the sedimentation phase. The duration of the sedimentation phase for default Cell Types can vary depending on the Cedex HiRes Software version used for the analysis. Sedimentation duration can be adjusted by the user in Cedex HiRes Software 2.x when creating or modifying a Cell Type.
\n
\nSedimentation duration can affect measurement results. It is important to check whether the Cell Types used for measurements have the same sedimentation duration, when comparing results between systems or software versions. For more information about adjusting the duration of the sedimentation phase, please see the chapter on adjustment of the image analysis using the Live Operator in the relevant Cedex HiRes Operator Guide.
\n
\nSensitive Dirt and Bubble detection
\nThe Dirt and Bubble dection in the Cedex HiRes Software is a very sensitive algorithm. Therefore, the algorith may in some cases detect dirt or bubble in the outlet or inlet of the measuring cell when no dirt or bubbles are actually present. A trained Roche Service Representative can adjust the position of the scan area. Experience has shown that adjusting the scan area improves the sensitivity of the dirt and bubble detection in the inlet and outlet regions. This adjustment has no influence on measurement results or the flow factor. Therefore, a validation of the system after adjusting the scan area is not necessary.
\nContact your local Roche representative for more information about this topic. 
\n ", "Name": "Troubleshooting - Help Corner" }, { "Language": "en", "Value": "Cedex HiRes Analyzer technology uses digital image recognition. The system uses Trypan Blue as the exclusion stain for cell viability determination. Digital image recognition technology enables to permanently store acquired measurement data.", "Name": "Principle" }, { "Language": "en", "Value": "Detectable range of concentration
\n
\nThe detectable concentration range for the Cedex HiRes System is 5x104 to 1x107 cells/ml.
\n
\nDescription of the Flow Factor
\n
\nThe flow factor is a conversion factor that is unique for each Cedex HiRes Analyzer flow chamber. Any time a flow chamber is removed and reinstalled or a new flow chamber is installed, the flow factor must be measured using the Cedex Density Reference Standard Beads because the chamber could be placed in a slightly different position, which can affect the conversion factor required for accurate measurements.
\n
\nCalculation of cell concentration
\n
\nThe calculation of cell concentration is based on a formula that involves the Flow Factor. Each system has a unique Flow Factor. Even when the actual cell count (i.e., number of cells measured) is similar for two systems, the final calculated concentrations may be different depending on the Flow Factor of each system. Note that the parameter, total cell count, cannot be used to compare results between systems. This parameter depends on the number of images selected by the user and the volume analyzed for each image. The final calculation of cell concentration also depends on the Flow Factor of each system.
\n
\nDimensional tolerance of the flow chamber
\n
\nAll cuvettes for the Cedex HiRes System have a chamber height of 0.1mm +/- 0.005mm. The thickness of every cuvette is interferometrically measured and certified with a tolerance of +/-0.00004mm. Each flow chamber is unique; therefore each Cedex HiRes Analyzer will have a flow chamber with a slightly different chamber height. The chamber height of the installed flow chamber is tracked in the System Options.
\n
\nChamber volume and number of images taken for a measurement
\n
\nThe total volume of the entire flow chamber is 11.25 μl. Only a portion of the chamber is used for taking images. For each image, 0.378 μl are analyzed in each image; 4.15 μl are analyzed when the precision is set to maximum (approximately 11 images).
\n
\nDue to slight variations in the chamber height of the flow chamber, the analyzed volume in one image will vary slightly from system to system.  Therefore, since precision is determined by the total volume analyzed, the number of images taken for a particular precision setting may vary slightly depending on the actual chamber height of the flow chamber installed in the system. The system will take as many images as necessary to reach the defined volume that should be analyzed for a particular precision setting.
\n
\nCalibration of the syringe
\n
\nThe syringe is a high precision liquid handling system using a stepper motor with a full stroke divided into several thousand steps.
\n
\nCedex Software Aggregate Rate and Aggregate Histogram
\n
\nAggregate Rate refers to the percentage of the total population of cells found in aggregates. If the Aggregate Rate is 70%, then only 30% of the cell culture is in a single-cell population. Aggregate Rate does not provide direct information about the size distribution of the aggregates. Information about the distribution of aggregate sizes is available using the Aggregate Histogram.
\n
\nFor the Cedex HiRes System, keys up to about 90 μm in diameter can enter the flow chamber. How many cells per aggregate this represents will depend on the average size of cells. For example, an aggregate of 15 cells with an average of 15 μm diameter per cell would have a diameter of approximately 75 μm and still pass through the flow chamber. Aggregates with more than 15 cells may be too large to pass through the flow chamber and, therefore, may be broken up into smaller aggregates.
\n
\nAggregates with more than 15 cells that are small enough to pass through the flow chamber can be analyzed, depending on the Live Operator parameter settings for the Cell Type chosen for the analysis of aggregates. In such a case, the total number of cells found in the aggregate will be displayed in the Aggregate Histogram, even if the number exceeds 15 cells per aggregate.
\n
\nAggregate counting
\n
\nAggregates are recognized using specialized image processing in the Cedex HiRes Operator Software algorithm. To properly analyze aggregates, the Cedex HiRes Operator addresses the following question: How to recognize small cells in aggregates versus large cells with internal structures as one cell?
\n
\nIn the Cedex HiRes System, the aggregate recognition algorithm acts on particles with an area within the size range from \"Aggregate Min. Diameter\" to \"Aggregate Max. Diameter\" set in the Live Operator. The appearance parameters, such as \"Viable Cells' Aggr. App.\" and \"Dead Cells' Aggr. App.\" in the Live Operator) control the segmentation process for live and dead cells. A higher setting of these parameters will direct the algorithm to segment areas which are then classified as viable or dead cells. Lower settings will do the opposite.
\n
\n
\nAssigning access rights for new user roles in the Cedex HiRes Software version 2.x
\n
\nUser roles for research, production, and support staff are preinstalled in the software, however individual rights and access to functions for these roles are not predefined. The Administrator must provide access rights for measuring, viewing the list, etc. to these roles before users assigned the roles can have access to any functions.
\n
\nThe Superuser and Adminstrator roles have functions that are assigned by default. Only the Administrator has access to the \"User and Rights Management\" function. The Superuser has access to all other software functions. Additional functions for user roles are optional and must be configured by a user role that has access to the \"User and Rights Management\" function. For more information about assigning access rights to users, refer to the relevant Cedex HiRes Analyzer Operator's Manual. The Operator's Manual is available online under \"Downloads\" tab of the Cedex HiRes Analyzer entry in the Online Technical Support.
\n
\nTrypan Blue storage conditions and use
\n
\nRoche recommends replacing the trypan blue with fresh trypan blue once per week. Trypan blue installed on the Cedex System for more than a week can form large particles due to precipitation, polymerization or contamination. These particles can enter the tubing and flow chamber, and potentially partially or completely block the inlet or outlet of the flow chamber.
\n
\nCalibration check recommendations using Cedex Density Reference Standard Beads
\n
\nThe Density Reference Standard Beads (DRSBs) mimic cell flow dynamics in the Cedex HiRes Analyzer. Their size and optical properties cause the Cedex Software to detect DRSBs as dead cells. Use the protocol in the Instructions for Use for DRSB to adjust the Cedex HiRes Analyzer calibration (flowfactor) or to verify the calibration.
\n
\nThere are no guidelines for how frequently to perform calibration checks using DRSB. Monthly calibration checks using 10 DRSB samples are a good starting point. However, each laboratory must establish their own DRSB calibration frequency based on their needs. Factors that should be considered include:\n
    \n\t
  • Risk analysis: Calibration testing using DRSBs is an after-the-fact check of the system. Such calibration checks confirm that the system was correctly calibrated between successful calibration checks. Such testing does not confirm whether the system will remain correctly calibrated for future measurements. Each lab must determine the acceptable risk before the next calibration check.
  • \n
\n \n\n
    \n\t
  • Laboratory and sample conditions: Dust and particles in a laboratory, and cell samples with large particles can affect Cedex HiRes Analyzer performance. Dust, large particles, and debris can enter the Cedex HiRes Analyzer tubing, resulting in clogging of tubing and the flow chamber. Clogging may not always be visible in the prescan image, and may require additional cleaning. Clogging can impact system calibration. For best results, perform frequent calibration checks when cell cultures have large particles, or dust and particles are present.
  • \n
\n
\nInformation about standard deviation values displayed in the Result Data table in the Measurement window
\n
\nThe Standard Deviation (Std Dev [cells/mL]) values displayed in the Result Data table in the Measurement window refer to the deviation from image-to-image of the total number of cells in each image. These results are expressed in terms of concentration. The Measurement Statistics Histogram also shows the total number of cells, viable or dead, in each image.
\n
\nLocation of images used for the Cedex HiRes Software image analysis
\n
\nAfter incubation with trypan blue, the stained cell suspension in the syringe is transported in one volume into the precision flow chamber. The flow chamber is scanned, and the scanned image is then partitioned into smaller images for evaluation. The number of images used for the evaluation depends on the number of images selected by the user for that measurement.
\n
\nSmaller images used for the analysis are selected from chamber locations in the following pattern: The first image is selected from the middle of the flow chamber. The next image selected is located below the first image, and third selected image is located above the first image. This pattern is repeated (middle image, followed by image below and image above) until the end number of images selected by the user is reached.
\n
\nChoosing a \"Priority\" for a newly created user
\n
\nWhen creating a new user, it is theoretically possible to choose different priorities for each user (\"High\", \"Normal\", or \"Low\"). With regard to priorities between different users, this function has no significant meaning at this time in software 2.x. 
\n
\nHowever, the system itself uses a priority \"Low\" for scheduling of, for example, cleaning routines (e.g., via the Scheduling function). Therefore, when measurements or Liquid Management (LM) functions are put into the queue by a regular user who has a priority of \"Normal\" or \"High\", the measurement or LM scheduled by the user will be processed before the processes scheduled by the system itself. If the user priority has been set to \"Low\", the system may first run system scheduled processes. In order to ensure that user scheduled measurements and processes always come first, one should always set the priority to \"High\" priority for users in order to always prioritize their actions over system actions.
\n
\nCalculation for the estimated number of runs left
\n
\nThe calculation for the estimated number of runs left, which is displayed at the top of the main Cedex Control Center window, is based on the actual amount of reagents used during the previous 25 fluidic actions (e.g., measurements or cleaning actions) performed on the system. If a very high number is displayed in this area, it is possible that not enough measurements or cleaning actions have been performed yet for an accurate calculation. 
\n 
\nThe estimated number of runs in the main window will be very high if a system or software is new, and less than 25 fluidic actions have been performed on the system. Once 25 fluidic actions have been performed, the estimated number of runs left will be calculated appropriately.
\n
\nIllumination
\nThe scanner in Cedex HiRes uses brightfield illumination.", "Name": "Background Information - Help Corner" }, { "Language": "en", "Value": "The Cedex HiRes Analyzer is a fully automated, image-based cell analyzer providing information about cell concentration, viability, aggregates, and morphological parameters, such as cell diameter and cell compactness. The analyzer uses the well-established Trypan Blue Exclusion Method and a high resolution image scanner to analyze cells and keys.

The Cedex Software features Live Operator for real-time optimization of the analysis algorithm, and a Cultivation Time Chart for monitoring cell growth to determine growth rates and doubling times. This powerful new technology enables applications such as accurate monitoring of baculovirus transfections via the measurement of cell diameter changes, making the Cedex HiRes Analyzer the first choice for your cellular research.

The Cedex HiRes Analyzer also represents the industrial standard for biopharmaceutical production. It is commonly used in GMP-validated biopharmaceutical production processes and conforms to GMP regulations, such as 21 CFR part 11.", "Name": "Product Description" }, { "Language": "en", "Value": "Additional reagents and equipment required to perform measurements
  • Cell culture grade Trypan Blue 0.2%
  • Cedex Detergent
  • Cedex Cleaning Solution
  • Distilled or deionized water
  • Sample cups, blue, transparent
  • Pipette
  • Pipette tips
", "Name": "Additional Equipment and Reagent Required" }, { "Language": "en", "Value": "Instrument Specifications
Dimensions: 40.5 cm x 53.5 cm x 46.5 cm (W x D x H)
Weight: 26.9 kg
Energy Requirements: 100 - 250 VAC, 50 - 60 Hz
Energy Consumption: 60 W
Method of Measurement: Digital image recognition
Detectable Cell Density Range: 5 x 104 - 1 x 107 cells/mL
Detectable Cell Diameter Range: 2- 40 μm
Required Sample Volume: 300 μL
Operating Temperature (optimal image quality is achieved between +20 to +30°C): +10 to +40°C (+50 to +100°F)
Maximum Temperature Change: +5°C/hour
Maximum Humidity Deviation: 5%/hour
Operating Humidity: 20-80% relative humidity (non-condensing)
", "Name": "Specification" }, { "Language": "en", "Value": "The technology of the Cedex HiRes Analyzer is based on digital image recognition. The system uses Trypan Blue as the exclusion stain for viability determination. The digital image recognition technology enables permanent storage of acquired measurement data.

System Features
  • Complete flow chamber check for full transparency of cell culture analysis
  • High-resolution scanner technology
  • Analysis of keys from 1 - 90 μm in size
  • Integrated High-Speed Multi Sampler for single samples and multiple runs
  • Audit trail for 21 CFR part 11 compliance
Software Features
  • Histograms
    • Diameter Histogram showing cell size distribution
    • Aggregate Histogram showing cell culture aggregation state
    • Compactness Histogram depicting cell morphology
  • Live Operator
    • High-speed real-time parameter optimization for specific cell lines
    • Customized analysis of acquired images
  • Scheduler
    • Automated cleaning and maintenance procedures
  • Database filter chart
    • Stores all results and images
    • Measurement results easily found using searchable directory
  • Drag and drop administration of users and user roles
    • User roles easily created for multiple or single users
    • Users can be added at any time with automatic update of user rights
", "Name": "Properties" } ] } } ] }

Cedex HiRes Analyzer

Cell culture analyzer

Cedex HiRes Analyzer
Cedex HiRes Analyzer
High resolution that counts
Cedex HiRes Analyzer - maximize performance with scanner-based imaging

  • Automated cell culture analysis: Save time and benefit from automated staining and mixing.
  • Integrated Data Management: Benefit from integrated data analysis and management capabilities.
  • Cultivation Time Chart: Compare growth curves from different cultures and calculate specific growth rate and doubling time.
  • Integrated System Suitability Test: Ensure high security of process quality for integration into GMP-validated environments.

 

For use in quality control / manufacturing process only.

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