Bio-rad S3™ Cell Sorter Manuel d'utilisateur

S3™ and S3e™ Cell SortersInstruction ManualCatalog #145-1001 #145 -10 02 #145-1005 #145-100 6 #145-1008

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Maintenance90 | S3 and S3e Cell Sorters7. Place a lint-free cloth in the nozzle chamber under the nozzle tip to prevent dropping the nozzle into t

10.4 Swap Nozzle Tip WizardInstruction Manual | 91Fig. 103. Nozzle swap wizard window.Fig. 102. Nozzle swap wizard window.Fig. 101. Nozzle swap wizard

92 | S3 and S3e Cell SortersMaintenanceFig. 105. Nozzle swap wizard window.20. Check to see if the stream is hitting the waste hole in the back of t

Instruction Manual | 9310.4 Swap Nozzle Tip WizardAn image of the stream alignment is shown with the stream camera. It is necessary to center the stre

94 | S3 and S3e Cell SortersMaintenanceThe window will give an option of repeating the gimball adjustment process or skipping and proceeding to drop

10.4 Swap Nozzle Tip WizardInstruction Manual | 95Fig. 110. Adjust Stream Position window pinhole viewFig. 111. Calibrating Droplets.Fig. 112. Calibra

96 | S3 and S3e Cell SortersMaintenanceFig. 113. Quality Control Process.10.4.2 Nozzle Tip Wizard with the AutoGimbal SystemThe AutoGimbal system us

Instruction Manual | 9710.4 Swap Nozzle Tip WizardFig. 114. AutoGimbal Swap Tip window.ABFig. 115. AutoGimbal Swap Tip window.

98 | S3 and S3e Cell SortersMaintenanceCleaning the nozzle tip:The nozzle tip may be clogged visibly, but there may also be a clog that is not visib

Instruction Manual | 9910.4 Swap Nozzle Tip Wizard20. Remove the lint-free cloth from nozzle chamber.21. Press the nozzle stage back down. The nozzl

Instruction Manual | 1Introduction1The S3™ and S3e™ Cell Sorters are designed to offer affordable, dependable, and simplified cell sorting. As benchtop

100 | S3 and S3e Cell SortersMaintenanceFig. 118. AutoGimbal Swap Tip window.Fig. 119. AutoGimbal Swap Tip window.

Instruction Manual | 10110.4 Swap Nozzle Tip Wizard26. If after reseating the nozzle tip the stream is still hitting the back wall, sort tray, or defl

102 | S3 and S3e Cell SortersMaintenanceFig. 122. Adjust Stream Position window.29. Using the Pitch and Roll arrows, adjust the stream’s position un

Instruction Manual | 10310.4 Swap Nozzle Tip Wizard38. The AutoGimbal system will commence automatic alignment of the nozzle tip and stream to the op

104 | S3 and S3e Cell SortersMaintenanceAfter the AutoGimbal system has completed its alignment and found the most optimal position, system calibrat

Instruction Manual | 10510.5 Optical Filter Cleaning10.5 Optical Filter CleaningThe optical filters in the S3 and S3e Systems will lose performance whe

Maintenance106 | S3 and S3e Cell SortersA decontamination wizard will start and walk through each procedure. When prompted follow the instructions.3

10.7 DecontaminationInstruction Manual | 1078. Replace the DI water and sheath fluid containers with at least 1 L of decontamination solution (Figure

108 | S3 and S3e Cell SortersMaintenanceAfter the system shuts down, the containers can be removed and the sheath and DI water cap assemblies should

10.7 DecontaminationInstruction Manual | 109Note: The user must be present to change the containers approximately 1 hour into the procedure. After thi

2 | S3 and S3e Cell SortersIntroductionThe S3 or S3e Cell Sorter complete system includes the following components:n S3 or S3e Cell Sorter Instrume

Maintenance110 | S3 and S3e Cell SortersAfter decontamination the system should be ready for running samples. Any high background should be eliminat

Instruction Manual | 111Troubleshooting11Table 30 describes possible errors and causes with troubleshooting steps. Table 30. Troubleshooting table. E

112 | S3 and S3e Cell SortersTroubleshootingTable 30. Troubleshooting table (continued). Error Possible Causes Troubleshooting Steps Drastic li

Instruction Manual | 113TroubleshootingTable 30. Troubleshooting table (continued). Error Possible Causes Troubleshooting Steps High event rate (

114 | S3 and S3e Cell SortersTroubleshootingTable 30. Troubleshooting table (continued). Error Possible Causes Troubleshooting Steps Decontamin

Instruction Manual | 115The following references and resources are useful for learning more about flow cytometry and its applications. Shapiro HM (2003

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Instruction Manual | 11713S3™ and S3e™ Cell Sorter SpecificationsThe specifications for the S3 and S3e Cell Sorters are shown in Table 31.Table 31. S3

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Instruction Manual | 119Ordering InformationCatalog # Description Instrumentation145-1001 S3 Cell Sorter (488 nm)145-1002 S3 Cell Sorter (488/561 n

Instruction Manual | 3This chapter describes the hardware of the S3™ and S3e™ systems. Understanding the system’s hardware is essential for proper ope

Life ScienceGroup14-1117 0614 Sig 121310031105 Rev D US/EGBio-Rad Laboratories, Inc.Web site www.bio-rad.com USA 800 424 6723 Australia 61

4 | S3 and S3e Cell SortersHardware Description2.1.1 Instrument Back PanelThe rear connector panel of the S3 or S3e system includes these features:

Instruction Manual | 52.2 Fluidics System2.2 Fluidics SystemThe S3 and S3e fluidics system consists of the bulk fluidics, loading stage, nozzle, and sor

6 | S3 and S3e Cell SortersHardware DescriptionThe sheath fluid and DI water are both filtered through a 0.2 µm filter to remove any particulates from

Instruction Manual | 72.2 Fluidics SystemDuring washing, a new sample can be loaded into the sample input station. The status of the loading stage is

Hardware Description8 | S3 and S3e Cell Sorters2.2.3 NozzleThe nozzle controls many crucial aspects associated with sorting such as:nCreating a sta

Instruction Manual | 92.3 OpticsThe sort positions for each collection vessel are numbered and will correlate to the position numbering in the softwar

10 | S3 and S3e Cell SortersHardware Description2.3.2 Beam Shaping Optic (BSO)The beam shaping optic sits in between the laser(s) and the interrogat

Instruction Manual | 112.4 Electronics2.4 ElectronicsThe S3 and S3e electronics process and deliver the emitted light signals to the software for user

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Instruction Manual | 13ProSort™ Software3This chapter describes the features of the ProSort Software. The software is the main interface for the S3™ o

14 | S3 and S3e Cell SortersProSort SoftwareTable 7. Control differences between administration and user modes. Control Administrator User Start

Instruction Manual | 153.1 Main Software Window3.1.1 Title Bar Buttons The title bar of the main software window provides the items listed in Table 8.

16 | S3 and S3e Cell SortersProSort Software3.1.3 Home Tab ToolbarThe Home tab toolbar consist of six sections for basic workspace functions and sor

Instruction Manual | 173.1 Main Software WindowTable 11. Region buttons and their functions (continued). Button Name Function Quadrant Adds a q

18 | S3 and S3e Cell SortersProSort Software3.1.4 Setup and Maintenance Tab ToolbarThe Setup and Maintenance tab toolbar consists of five sections (F

Instruction Manual | 193.1 Main Software WindowTable 17. Fluidics buttons and their functions. Button Name Function Debubble Removes bubbles fr

Instruction Manual | iCopyright © 2014 by Bio-Rad Laboratories, Inc. All rights reserved. Toll Free: 1-800-4-BIORAD (1-800-424-6723)Fax: 510-741-5800F

20 | S3 and S3e Cell SortersProSort Software3.2 Control PanelSystem operation is performed and monitored with the control panel (Figure 16). The con

Instruction Manual | 213.2 Control PanelTable 20. Instrument controls and their functions. Button Name Function Start Sample Starts sample acqui

22 | S3 and S3e Cell SortersProSort SoftwareFig. 17. Administrator tab toolbar.Table 20. Instrument controls and their functions (continued). Button

Instruction Manual | 233.3 Administrator Tab ToolbarTable 22. Administrator tab toolbar buttons and their functions. Button Name Function Edit

24 | S3 and S3e Cell SortersProSort SoftwareFig. 18. Edit QC Criteria window with green plus button and rewind arrow highlighted.3.3.2 Edit Drople

Instruction Manual | 253.3 Administrator Tab ToolbarFig. 19. Droplet Controls window with drop settings highlighted.Calibrate dropletsRecalculate drop

26 | S3 and S3e Cell SortersProSort Software3.3.4 User ReportsUser reports can be viewed, saved, printed, and exported for analysis of system usage

Instruction Manual | 273.3 Administrator Tab ToolbarFig. 23. Manage Users window.3.3.5 Managing UsersThe Manage Users button opens the Manage Users wi

ProSort Software28 | S3 and S3e Cell Sorters Fig. 25. Sample Calibration Warning dialog box.Fig. 26. Calibration Sample Offset window. 3.3.6 Deconta

Instruction Manual | 293.3 Administrator Tab Toolbar3.3.8 Global PreferencesUsers with administrator privileges have the ability to change global sett

ii | S3 and S3e Cell SortersBio-Rad Laboratories ResourcesBio-Rad provides many resources for scientists. Table 1 lists available resources and how

30 | S3 and S3e Cell SortersProSort SoftwareBoost AdjustmentThe default boost duration is optimized to bring the sample into position for interrogat

3.3 Administrator Tab ToolbarInstruction Manual | 31Parameter NamesEach parameter is defaulted to its channel name (FSC, SSC, FL1, FL2, FL3, and FL4).

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Instruction Manual | 33Getting Started4After system installation by a Bio-Rad certified service engineer, the S3™ or S3e™ System should not be turned o

34 | S3 and S3e Cell SortersGetting StartedIMPORTANT! n When handling sheath fluid and DI water containers, minimize air exposure to help avoid con

Instruction Manual | 354.3 Daily StartupOn a daily basis, the system requires approximately 20 min of warmup time before running quality control or an

36 | S3 and S3e Cell SortersGetting StartedDuring the startup process, the fundamental alignment window will also open to initiate cameras, maintain

Instruction Manual | 374.4 Quality ControlNote: If the bead lot is not available in the dropdown menu, refer to Section 3.3.1, Editing QC Criteria. A

38 | S3 and S3e Cell SortersGetting Started4.4.2 Side StreamsFollowing droplet setup, the side streams are adjusted to align with the sort tube posi

Instruction Manual | 394.4 Quality ControlFig. 39. QC histograms displayed in the workspace.4.4.3 AlignmentDuring the alignment phase, automated micro

Instruction Manual | iiiSafety and Regulatory ComplianceFor safe operation of the S3 and S3e Cell Sorter systems, we strongly recommend that you follo

40 | S3 and S3e Cell SortersGetting Started4.4.5 Drop Delay MaintenanceAfter drop delay determination, the system will be automatically placed into

Instruction Manual | 414.4 Quality Control4.4.6 Recalculate Drop DelayTo recalculate the drop delay using ProLine calibration beads, click the Recalcu

42 | S3 and S3e Cell SortersGetting Started4.5 Protocols and WorkspaceBefore acquiring samples, a protocol with at least one histogram or density pl

Instruction Manual | 434.5 Protocols and Workspace4.5.2 Loading Instrument SettingsProtocols can be saved for future use. Each protocol is saved with

44 | S3 and S3e Cell SortersGetting Started4.5.4 Displaying Multiple ProtocolsMultiple protocols can be opened at the same time by opening an existi

Instruction Manual | 454.5 Protocols and Workspace4.5.8 Histograms and Density PlotsHistograms and density plots can be created from the Home tab. Cli

46 | S3 and S3e Cell SortersGetting StartedPlots can be moved within the workspace by grabbing the bar at the top and dragging to the desired positi

Instruction Manual | 474.5 Protocols and Workspace4.5.11 Viewing StatisticsTo view the statistics on a plot, highlight the plot and select the Statist

Getting Started48 | S3 and S3e Cell Sorters1. Right click on the Statistics panel.2. Click on the statistic name and drag and drop to the new locati

Instruction Manual | 494.5 Protocols and WorkspaceA region can be moved, tilted, resized, and deleted (Figure 52). 1. Click the desired region.2. To

iv | S3 and S3e Cell SortersSafe Use Specifications and ComplianceLaser Product Hazard ClassificationThe intent of the laser hazard classification is t

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Instruction Manual | 51Sample acquisition can be done independent of sorting. As a sample is run through the system, data are gathered and can then be

52 | S3 and S3e Cell SortersAcquisitionNote: Cycle mode can be enabled or the refresh button can be used to refresh the data displayed on the screen

Instruction Manual | 535.1 Aquisition Setup5.1.2 Event LimitAn event limit may be set on the acquisition (Figure 55). This limit will be based on the

54 | S3 and S3e Cell SortersAcquisition5.1.7 Auto SaveAn auto save option is available as a check box on the instrument control panel. If this box i

Instruction Manual | 555.2 Compensation5.1.8 Pausing and ResumingAcquisition may be paused and resumed while in progress using buttons on the instrume

56 | S3 and S3e Cell SortersAcquisitionThe same can be done for the FL2 positive control, though the compensation required is much less (Figure 60)

Instruction Manual | 575.2 Compensation5.2.1 Auto Compensation WizardThe compensation wizard assists with establishing the proper compensation matrix

58 | S3 and S3e Cell SortersAcquisition6. Locate and select the single control FCS file for each parameter.7. Select whether to use height or are

Instruction Manual | 595.2 Compensation5.2.2 Manual CompensationIt is advised to use the auto compensation feature of the software, however it is poss

Instruction Manual | vn Wash your hands thoroughly with soap and water after working with any potentially infectious material before leaving the labo

60 | S3 and S3e Cell SortersAcquisition5.3 Checking or Swapping FluidicsThe bulk fluidic containers are located inside the system and can be accessed

Instruction Manual | 613. Open fluidics door.4. Detach the quick disconnect from the cap assembly by pushing the metal button.5. Lift the quick disc

Acquisition62 | S3 and S3e Cell Sorters11. Place the container back into position.12. Attach the quick disconnect to the cap assembly. An audible c

Instruction Manual | 63In front of each PMT is a holder for a neutral density filter or bandpass filter. Each filter holder contains a screw-in retaining

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Instruction Manual | 65Sorting6Sorting cell populations is highly dependent on several factors such as cell type, population frequency in the starting

Sorting66 | S3 and S3e Cell Sorters14. Close the sort chamber door. 15. Click Start Sort on the instrument control panel. The Sort Statistics win

Instruction Manual | 676.1.1 Sort CollectionThe S3™ and S3e™ Systems offer four types of sort collection vessel options:n5 ml tubesn1.5 ml tubesn8-

68 | S3 and S3e Cell SortersSortingNote: If a sort limit is selected and volume tracking enabled, whichever criterion (limit or full tube) is met fir

Instruction Manual | 696.2 Sort Modes6.2 Sort ModesTable 24 describes the three sort modes available when setting up a sort. Sort modes can be selecte

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70 | S3 and S3e Cell SortersSortingTable 25 describes the available sort statistics. Each statistic is sort-stream specific. The Sort Statistics wind

Instruction Manual | 716.3 Sort Statistics6.4 Sort PlotsDuring a sorting experiment a sort plot can be created in the current protocol. This plot dis

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Instruction Manual | 73Additional Software Features 7The ProSort™ software is an acquisition and sorting software package with the additional features

74 | S3 and S3e Cell SortersAdditional Software Features7.4 Clean SystemIn between samples or users, cleaning can be done at low or high pressure. H

Instruction Manual | 757.5 Instrument Status BoxThe cleaning protocol will be completed when the window disappears. If a high pressure cleaning has be

76 | S3 and S3e Cell SortersAdditional Software Features7.8 Quality Control ReportsTwo different reports are available to track the daily performanc

Instruction Manual | 777.9 User ReportsUser reports are accessible by administrators and can be used to track system usage over time (Figure 86). Thes

Additional Software Features78 | S3 and S3e Cell SortersIMPORTANT! The S3 Biosafety System must remain on at all times when the S3 or S3e Cell Sorte

Instruction Manual | 79Shutdown88.1 Daily ShutdownThe S3™ and S3e™ System should be shut down at the end of the day. This will turn off all droplet fo

Instruction Manual | viiTable of ContentsChapter 1: Introduction 11.1 System Components 21.2 Installation Requirements 2Chapter 2: Hardware Descrip

80 | S3 and S3e Cell SortersShutdown3. Click Yes to confirm shutdown.Note: If there are any unsaved protocols, the software will warn you that uns

Instruction Manual | 818.1 Daily ShutdownA window will open requesting a tube of cleaner to be used to clean the system (Figure 91). 6. Load a tube o

Shutdown82 | S3 and S3e Cell SortersOnce the system is clean, the shutdown procedure will shutdown hardware, fluidics, air will be purged, rinsing an

headerInstruction Manual | 839Automatic Startup The ProSort™ can schedule an automatic startup of the system. Upon automatic start up, the system will

84 | S3 and S3e Cell SortersAutomatic Startup9.2 Previously Scheduled Automatic StartupsAfter double clicking the ProSort Software icon on the deskt

Instruction Manual | 859.2 Previously Scheduled Automatic Startups When a user logs onto the system and an automatic startup time is designated, the s

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Instruction Manual | 87Maintenance Always follow the PPE guidelines relevant to your laboratory’s safety procedures for dealing with the chemicals re

88 | S3 and S3e Cell SortersMaintenance10.2 Dealing with ClogsA clogged or partially clogged nozzle tip can occur while analyzing or sorting and it

Instruction Manual | 8910.4 Swap Nozzle Tip Wizard10.4.1 Nozzle Tip Wizard with the Agilis SystemThe Agilis system uses two axis-motorized positioners