BioFrac™ Fraction Collector User Guide Version 2.
BioFrac™ Fraction Collector User Guide Version 2.
Bio-Rad Technical Support Department The Bio-Rad Technical Support department in the U.S. is open Monday through Friday, 5:00 AM to 5:00 PM, Pacific Standard Time. Worldwide technical support is available on the Web at www.consult.bio-rad.com. Phone: 1-800-424-6723, option 2 Fax: 1-510-741-5802 Email: LSG.TechServ.US@Bio-Rad.com (U.S.) LSG.TechServ.Intl@Bio-Rad.com (International) Web: www.consult.bio-rad.
Table of Contents Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Physical Description . . . . . . . . . . . . .
Table of Contents The Run Screen in LP/Econo Mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Rack Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Method Library Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Calibration Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Advanced Collection Screen . . . . . . . . . . . . . . . . . . .
Table of Contents Appendix C Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Appendix D Warranty and Ordering Information . . . . . . . . . . . . . 93 Warranty Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of Contents vi | BioFrac Fraction Collector
1 Introduction Overview The BioFrac™ fraction collector provides automated collection options for analytical and preparative chromatography applications. It can be used as a stand-alone collector or as a companion to any chromatography system. The BioFrac fraction collector is capable of performing basic to complex fraction collection schemes and can be used at flow rates up to 100 ml/min. The fraction collector accommodates numerous rack options from microplates to bottles and carboys.
1 | Introduction Fig. 1. BioFrac fraction collector with two F1 racks (12–13 mm tubes).
Overview Advanced fraction collection functions.
1 | Introduction Optional drop former optimized for small-volume drop dispensing (recommended when collecting into microplates) Ability to start/stop an external pump and chart recorder Screen sleep mode for longer display life Unpacking When unpacking the fraction collector, carefully inspect the containers for any damage that may have occurred in shipping. Severe damage to a container may indicate damage to its contents.
Overview Fig. 2. Fittings kit. LEGEND Item Quantity Contents of Fittings Kit 1 1 1.0 m Tefzel tubing, 1/16" OD, 0.030" ID, with one 1/4-28 fitting 2 4 Fittings, 1/4-28, 1/16" OD 3 4 Ferrules, 1/16" OD 4 2 Unions, luer to 1/4-28 5 1 26” PEEK tubing, 1/16" OD, 0.030" ID, with two 1/4-28 fittings labeled collect HF 6 1 26” PEEK tubing, 1/16" OD, 0.
1 | Introduction Physical Description Figure 3 identifies the physical features of the BioFrac fraction collector, and the accompanying table describes the features in greater detail. Fig. 3. Physical features of the BioFrac fraction collector.
Overview Feature Description Control module The fraction collector head contains the display, function keys, and alphanumeric keypad (see Chapter 2, Front and Rear Panel Controls and Connectors, on page 9). Its height may be adjusted to accommodate tube heights up to 150 mm. Base The fraction collector base holds the rack positioning tray, power switch, and I/O connectors.
1 | 8 | Introduction Feature Description Drophead Consists of the drop former, a photodiode cell for drop counting, and a clear glass tube that protects the photodiode cell from splashes. The inlet tubing is connected to the drop former, which provides uniform drop size. The drop former accepts 1/4-28 fittings. An optional drop former is available that has been optimized for small-volume drop dispensing (25 µl drops). Diverter valve The diverter valve minimizes spillage during drophead movement.
2 Front and Rear Panel Controls and Connectors The controls and connectors on the front and rear of the panel are illustrated and described in this chapter. Front Panel Controls Fig. 4. Front panel controls.
2 | Front and Rear Panel Controls and Connectors Table 1. Front panel controls Feature Description Cursor keys Used to move the cursor on the LCD display up, down, left, or right. Function keys These five keys are located directly below the LCD display. The function each key executes is displayed above it. Clear Entry key Clears a cursor field or closes a menu option list. Decimal point key Used to enter a decimal point. Enter key Accepts a numeric value that has been entered.
Rear Panel Connectors Table 2. Rear panel connectors Connector Description Display cable – connects the fraction collector control module to the power supply and other ports in the base of the fraction collector. Diverter valve connector – used to connect the diverter valve to the fraction collector. I/O connector – a 15-pin D connector used for connecting the following instruments to the BioFrac™ fraction collector: BioLogic™ LP chromatography system, Model EP-1 Econo™ pump.
2 | Front and Rear Panel Controls and Connectors Table 2. Rear panel connectors, continued. Connector Description Rec connector – this 8-pin mini-DIN connector controls the diverter valve or a chart recorder: When connected to a BioLogic LP system or Model EP-1 Econo pump, the connector receives diverter valve control signals. When connected to a chart recorder, the fraction collector controls paper feed, pen up/down, and event marks.
3 System Configuration and Plumbing The BioFrac™ fraction collector is shipped assembled and requires minimal plumbing and cabling to prepare it for use.
3 | System Configuration and Plumbing Fraction Collector Setup Fig. 6. Plumbing the BioFrac fraction collector.
Fraction Collector Setup To set up the BioFrac fraction collector 1. Place the fraction collector on a level surface on a laboratory or coldroom bench or in a cold cabinet. Alternatively, if benchspace is limited, the modular fraction collector can be stacked on top of the NGC, BioLogic DuoFlow™, or BioLogic™ LP chromatography system workstations. 2. Connect the fraction collector display cable to the port labeled Display on the back of the fraction collector. (See Figure 5 on page 10.) 3.
3 | System Configuration and Plumbing 8. Attach the tubing from your chromatography system to the Common port on the diverter valve using either a 1/4-28 fitting or luer to 1/4-28 union. The BioFrac fittings kit includes 1/4-28 fittings (for 1/16" OD tubing) and luer to 1/4-28 unions. 9. Connect the fraction collector to your chromatography system as described in the next section, Connecting Instruments and Devices to the BioFrac Fraction Collector.
Connecting Instruments and Devices to the BioFrac Fraction Collector of fraction collection. Devices not manufactured by Bio-Rad must be capable of transistor to transistor logic (TTL) control. Note: Before proceeding, make sure power to each component to be connected is turned off. Be sure to use a grounded, surge-protected outlet when plugging in the power cables.
3 | System Configuration and Plumbing To connect the fraction collector to a BioLogic DuoFlow system 1. Connect the bus cable to the connector marked INSTRUMENT BUS on the base of the fraction collector. 2. Connect the other end of the bus cable to either the INSTRUMENT BUS connector on the back of the BioLogic DuoFlow workstation or to other Bio-Rad components that are daisy chained to the BioLogic DuoFlow workstation by the instrument bus. 3.
Connecting Instruments and Devices to the BioFrac Fraction Collector To connect the fraction collector to the BioLogic LP system or Model EP-1 Econo pump 1. 2. Connect system cable 15: a. Connect the cable’s 15-pin D connector to the port labeled I/O on the fraction collector. b. Connect the mini-DIN connector to the port labeled Fraction Collector on the BioLogic LP system or the port labeled on the Model EP-1 Econo pump. Connect system cable 3: a.
3 | System Configuration and Plumbing Connecting to the Econo Gradient Pump and a Chart Recorder The following configuration shows the fraction collector set up with an Econo gradient pump and a chart recorder. This system allows full use of the fraction collector’s advanced programming features. Fig. 9. Connecting the fraction collector, Econo gradient pump, and chart recorder. To connect the fraction collector, Econo gradient pump, and chart recorder 20 1.
Connecting Instruments and Devices to the BioFrac Fraction Collector Connecting the Model EP-1 Econo and Econo Gradient Pumps The BioFrac fraction collector can start an Econo gradient pump or Model EP-1 Econo pump remotely. To make the necessary connections you will need a BioFrac accessory cable (catalog # 731-8290) and system cable 7 (catalog # 731-8267). 1. Connect the BioFrac accessory cable green/black and blue/white wires to the system cable 7 blue wire. 2.
3 | System Configuration and Plumbing collector to another manufacturer’s pump, UV monitor, and/or chart recorder, you need a BioFrac accessory cable (15-pin to bare wires), system cable 7 (mini-DIN to bare wires), and 2-wire shielded cable (26 gauge or larger, available from most electronics or hardware stores). In addition, the pump and chart recorder circuit logic must be Start/closed circuit = ON; Stop/open circuit = OFF. The UV monitor’s analog output must be 100 mV or 1 V.
Connecting Instruments and Devices to the BioFrac Fraction Collector 3. Connect the BioFrac accessory cable green/black wire (pin #9) to the start pin on the remote instrument. Refer to your particular pump’s documentation for further information. 4. Connect the orange/black wire (pin #10) to the remote instrument’s signal ground. When Run is pressed on the BioFrac instrument, the remote start relay pins (pins #9 and #10) are connected and the start signal is relayed to the remote device.
3 | System Configuration and Plumbing To remotely control the drophead and diverter valve 1. Set up the fraction collection parameters as described in Chapter 6, LP/Econo Mode Operation, on page 75. In LP/Econo mode, the remote instrument controls all aspects of fraction collection. 2. Connect a BioFrac accessory cable (15-pin to bare wires; see Appendix B, Rear Panel Connector Information, on page 85) to the I/O port on the back of the fraction collector. 3.
Connecting Instruments and Devices to the BioFrac Fraction Collector Setting Up the Fraction Collector Racks The BioFrac fraction collector is designed to accommodate a variety of rack options, including both custom-molded and off-the-shelf racks. Custom racks include the ice bath/microplate and Prep-20 racks. The top of the ice bath rack holds microplate (12-, 24-, 48-, and 96-well) and Titertube tube racks that adhere to Society of Biomolecular Screening (SBS) standards for microplates.
3 | 26 System Configuration and Plumbing | BioFrac Fraction Collector
Connecting Instruments and Devices to the BioFrac Fraction Collector User Guide | 27
3 | System Configuration and Plumbing Table 3. BioFrac fraction collector racks Rack Capacity Format Column x Rows Tubes Per Rack (Total) Rack ID Bio-Rad Catalog # F1 741-0010 Holds 12–13 mm diameter tubes up to 100 mm in height 2 racks 6 x 15 90 (180) F2 741-0011 Grip rack, holds 15–16 mm diameter tubes up to 150 mm in height 2 racks 5 x 12 60 (120) F3 741-0012 Grip rack, holds 18–20 mm diameter tubes up to 150 mm in height 2 racks 4 x 10 40 (80) H1 741-0013 For 1.5–2.
Connecting Instruments and Devices to the BioFrac Fraction Collector Table 3. BioFrac fraction collector racks, continued. Rack ID Bio-Rad Catalog # Rack Description Rack Capacity Format Column x Rows Prep-20 741-0018 Preparative rack 1 2 x 10 For 250 ml bottles 1 Bottle Tubes Per Rack (Total) 20 funnels 4 bottles Other racks compatible with the BioFrac fraction collector are available from Scienceware (www.belart.com).
3 | System Configuration and Plumbing Ice Bath Remove the lid of the ice bath/microplate rack and fill the tub approximately ½ full with crushed ice, replace the lid and insert 13 x 100 mm culture tubes. Remove the rack positioning tray from the fraction collector and replace it with the ice bath rack. Adjust the fraction collector control module appropriately for the height of the tubes being used.
Connecting Instruments and Devices to the BioFrac Fraction Collector To install the Prep-20 adaptor, remove the two green rubber plugs on the left, front and right, front of the fraction collector. After attaching the Tygon tubing, insert the Prep-20 adaptor into the preparative rack holder slots. Notice that the drain trough slopes slightly towards the drain funnel. The collection port tubing extends down the front of the fraction collector.
3 | System Configuration and Plumbing To adjust the drophead height 1. Disengage the lock by pulling both lock pins outward from the fraction collector. The lock pins can be disabled by turning the handles sideways (Figure 11). 2. Move the control module to the desired height (Figure 12). 3. Move the control module up or down until the lock pins click into position. Fig. 11. Lock pin adjustment.
Connecting Instruments and Devices to the BioFrac Fraction Collector Fig. 12. Height adjustment.
3 | 34 System Configuration and Plumbing | BioFrac Fraction Collector
4 The User Interface Control of the fraction collector interface is through the arrow, ENTER, CLEAR, and alphanumeric keys that are located to the right of the display or through the function keys located below the display. Fig. 13. BioFrac™ fraction collector user interface. Navigation between the fields is accomplished using the arrow keys. Numbers and/or letters are entered into the alphanumeric fields using the alphanumeric keypad. Use ENTER to accept the changed parameter.
4 | The User Interface A, B, C, 2, and so on). The five function keys are used to move between different screens or perform predefined functions. When the cursor is in a menu selection field, pressing ENTER causes the menu to be displayed. Once a menu is displayed, the up/down arrow keys are used to navigate through the menu items. Pressing ENTER selects the menu item shown at the cursor position.
Main Screen Table 4. Main screen parameters (Local mode) Parameter Function Method Displays the current method name (text only). Collect Displays the currently selected collection mode: All, Threshold, Windows, Windows/Threshold (text only). Rack Displays the currently selected rack (text only). MultiRun Menu for choosing the multiple run function: Overlay – causes collection to occur in the same tubes for each subsequent experiment.
4 | The User Interface Table 4. Main screen parameters (Local mode), continued. Parameter Function min/ml/drop Menu for choosing the fraction size units, time (min), volume (ml), drops. Note: If collecting by volume, the flow rate must be entered in the flow rate field. Flow Rate Used to define the flow rate. Must be set if collecting by volume. This field is not displayed if collecting in time or drop mode. Table 5.
Main Screen Main Screen (LP/Econo Mode) In LP/Econo mode, the Main screen is used to set the multirun mode and start and end tube numbers (see Figure 15, Table 6, and Table 7). All other fraction collection parameters are controlled remotely. Function keys are used to engage the fraction collector so that it will accept remote divert and fraction advance signals (see LP/Econo Mode) or to select a rack from the rack screen. Fig. 15. Main screen (LP/Econo mode). Table 6.
4 | The User Interface Table 6. Main screen parameters (LP/Econo mode), continued. Parameter Function MultiRun Menu for choosing the multiple run function: Overlay – causes collection to occur in the same tubes for each subsequent experiment. In this mode, the fraction collector moves back to the start tube when the end tube is filled. Stop is used to disengage the experiment. Seq. Tube+1 – increments the start and end tube numbers at the end of each run so that one tube is skipped between runs.
Main Screen Table 7. Main screen function keys (LP/Econo mode) Function Key Function Engage Moves the drophead to the start tube and causes the fraction collector to listen for fraction advance and diverter valve signals. Rack Displays the Rack Selection screen.
4 | The User Interface Run Screen The Run screen provides information about the progress of the current run. The information displayed on the screen depends on the type of method being used and whether the fraction collector is in Local or LP/Econo mode. The Run Screen in Local Mode In Local mode, the information displayed includes run status, run time, fraction filled, fraction size, and drophead position.
Run Screen Table 8. Run screen parameters (Local mode) Parameter Function Status Displays the current fraction collection status: Collecting – the diverter valve is in the collect position and fractions of a user-specified size are being collected. Diverting – the diverter valve is in the Waste position. Run Paused!: The run is currently paused and the diverter valve is at Waste. A warning is displayed that the diverter valve is diverting to Waste.
4 | The User Interface Table 9. Run screen function keys (Local mode) 44 Function Key Function Stop Stops the current experiment and displays the Results screen. Advance Advances the drophead by one tube. Divert/Collect Toggles the diverter valve between Collect and Waste. Pause Pauses fraction collection and switches the diverter valve to Waste during the pause. Resume Resumes fraction collection after a pause.
Run Screen The Run Screen in LP/Econo Mode In LP/Econo mode the Run screen shows the current diverter valve status and drophead position (see Figure 17 and Table 10). This screen has a Stop function key that causes the fraction collector to stop listening for fraction advance and diverter valve signals and returns the drophead to the home position. Fig. 17. Run screen (LP/Econo mode). Table 10.
4 | The User Interface Rack Screen The Rack screen is used to select the rack type and collection pattern to be used in a method (see Figure 18, Table 11, and Table 12). The default collection pattern is serpentine; however, it may be changed to collection by row or by column when collecting in microplates or Titertube tubes. The Divert Between Tubes feature can be used to reduce the amount of liquid spilled during tube advances.
Rack Screen Table 12. Rack screen function keys Function Key Function Done Accepts all changes and returns to the Main screen. Cancel Aborts all changes and returns to the Main screen.
4 | The User Interface Method Library Screen The Method Library screen is used to retrieve, save, or delete user-defined collection methods (see Figure 19 and Table 13). Up to 20 methods may be stored. Function keys are used to load, save, and delete methods or to get to the Calibration screen (see Calibration Screen on page 49). Fig. 19. Method Library screen. The Method Library displays the Default and user-defined methods. Table 13.
Calibration Screen Calibration Screen Use the Calibration screen to adjust the drophead calibration, screen contrast, sleep mode, and to zero the analog-to-digital converter (AtoD). Refer to Table 14 for more information about the use of each function key. Table 14. Calibration screen function keys Function Key Function Done Returns the display to the Method Library screen. Contrst Allows the user to set the display contrast.
4 | The User Interface Table 14. Calibration screen function keys, continued. Function Key Function Sleep Allows the user to select how many minutes the display should wait before going into sleep mode. Pressing any button will wake up the display. Calibrating the Fraction Collector X–Y Arm To calibrate the Fraction Collector X–Y arm Caution: Using this function overwrites the previous calibration. 50 1. Remove the drop former top by twisting it counterclockwise and gently lifting it out. 2.
Advanced Collection Screen Advanced Collection Screen The Advanced Collection screen is used to turn threshold and windows collection on or off and to set threshold and delay parameters (see Figure 20, Table 15, and Table 16). Threshold parameters are displayed only when threshold is turned on. From this screen, the threshold level and UV detector input voltage (100 mV or 1 V) can be entered and the user can specify whether non-peak fractions are to be collected or not.
4 | The User Interface Table 15. Advanced Collection screen parameters, continued. Parameter Function %AUFS Current threshold setting. (Displayed only when Threshold is on). Full Scale Menu for defining the UV detector’s output voltage. (Displayed only if Threshold is on.) The detector input voltage can be set to either 100 mV or 1 V. NonPeak Frac. Turns collection of non-peak fractions on or off. (Displayed only if Threshold is on). Size The non-peak fraction size.
Advanced Collection Screen Table 16. Advanced Collection screen function keys Function Key Function Done Accepts all changes and returns to the Main screen. Cancel Aborts all changes, including changes made in the Collection Windows Table screen (see the Table function key). Table Changes the screen to the Collection Windows Table screen (see Collection Windows Table Screen on page 54). (Visible only when Windows is on).
4 | The User Interface Collection Windows Table Screen The Collection Windows Table screen is used to enter the collection windows parameters (see Figure 21, Table 17, and Table 18). Up to 20 different collection windows can be defined, each with a different fraction size and threshold. The threshold parameter column (Thold) shown in Figure 21 is displayed only when threshold collection has been turned on in the Advanced Collection screen (see Advanced Collection Screen on page 51).
Collection Windows Table Screen Table 17. Collection Windows Table screen parameters, continued. Parameter Function FrcSz The fraction size for the current window (min, ml, drops). The header for this column is also a menu that can be used to change the fraction size units to min, ml, or drops. Thold The threshold value for each collection window. Table 18.
4 | The User Interface Results Screen The Results screen provides a list of the tubes associated with each peak or window. When collecting by threshold, a peak is defined as the tubes collected while the UV signal was above threshold. The Results screen is displayed at the end of each run, if the run was not started remotely, or can be viewed by pressing F5 on the Main screen (see Main Screen on page 36).
5 Stand-Alone Operation Normal operation of the BioFrac™ fraction collector is in Local mode (stand-alone mode). As a stand-alone fraction collector, the BioFrac device may be used to collect fractions based on time (minutes), volume (milliliters), or drops (up to a flow rate of 5.0 ml/min). In Local mode, the fraction collector controls all aspects of fraction collection and is not in communication with Bio-Rad’s BioLogic DuoFlow™ chromatography system, BioLogic™ LP system, or Model EP-1 Econo™ pump.
5 | Stand-Alone Operation Windows Collection — enables you to specify periods of time or volumes (windows) during which fractions are to be collected. For example, a window can be defined to start after an initial void volume. The BioFrac fraction collector lets you define up to 20 different time or volume windows. The liquid delivered during a collection window is collected into tubes, whereas the liquid delivered outside of a collection window is diverted to waste.
Collect All Collect All Fig. 23. Running a Collect All method. To run a Collect All method 1. On the Main screen, ensure that Collect is set to Collect –> All. If the screen is not set to Collect –> All, press Collect (F4) and turn Windows and Threshold off. Alternatively, press Method (F3) and load the method Default. 2. To select a rack, press Rack (F2) and choose a rack from the Rack menu. Choose whether you want the diverter valve to divert to waste during fraction advances.
5 | Stand-Alone Operation The fraction collector stops when the last tube is reached unless a stop command is received before it reaches the last tube. At the end of a run, the start tube number and end tube number will be automatically updated according to the MultiRun mode selected. 5. Set the fraction size units (min, ml, or drops) and enter the fraction size. If collecting by volume, you must enter a flow rate. 6.
Peak Detection by Threshold Peak Detection by Threshold Fig. 24. Running a Collect by Threshold method. To run a Collect by Threshold method 1. To select a rack, press Rack (F2) and then choose a rack from the Rack menu. Choose whether or not you want the diverter valve to divert to waste during fraction advances. If you are collecting in microplates or Titertube tubes, choose a collection pattern: Serpentine, Row, or Column.
5 | Stand-Alone Operation The fraction collector stops when the last tube is reached unless a stop command is received before it reaches the last tube. At the end of a run, the start tube number and end tube number are automatically updated according to the MultiRun mode selected. 4. Set the fraction size units (min, ml, drops) and set the fraction size. If collecting by volume, you must enter a flow rate. 5. Press Collect (F4) and turn Threshold on.
Collect by Windows Collect by Windows Fig. 25. Running a Collect by Windows method. To run a Collect by Windows method 1. To select a rack, press Rack (F2) and then choose a rack from the Rack menu. Choose whether you want the diverter valve to divert to waste during fraction advances. 2. (Optional) If you are collecting in microplates or Titertube tubes, choose a collection pattern: Serpentine, Row, or Column. 3.
5 | Stand-Alone Operation number and end tube number are automatically updated according to the MultiRun mode selected. 6. Set the fraction size units (min, ml, or drops) and set the fraction size. If collecting by volume, you must enter a flow rate. 7. Press Collect (F4) and turn Windows on. 8. Press Table (F3) and enter the windows and fraction size in the Collection Windows Table screen. Note that the fraction size entered in the Main screen is the default size.
Collect by Windows and Threshold Collect by Windows and Threshold Fig. 26. Running a Collect by Windows and Threshold method. To run a Collect by Windows and Threshold method 1. To select a rack, press Rack (F2) and then choose a rack from the Rack menu. Choose whether you want the diverter valve to divert to waste during fraction advances. If you are collecting in microplates or Titertube tubes, choose a collection pattern: Serpentine, Row, or Column.
5 | Stand-Alone Operation number and end tube number are automatically updated according to the MultiRun mode selected. 4. Set the fraction size units (min, ml, drops) and set the fraction size. If collecting by volume, you must enter a flow rate. 5. Press Collect (F4) and turn Threshold on. Set the global %AUFS (absorbance units full scale) and set the detector input voltage to either 100 mV or 1 V full scale depending on your detector’s output voltage. Press Done.
Collect Using a Delay Function Collect Using a Delay Function The delay volume is the volume of fluid contained in the path between the significant detector, typically the UV monitor, and the fraction collector drophead. When the system is plumbed, there is typically a length of tubing between the detector(s) and the fraction collector. Fluid in the path of the detector must pass through this tubing to arrive at the fraction collector. Thus the length of the tubing defines the delay volume.
5 | Stand-Alone Operation Table 20. Volume of commonly used tubing and inline devices, continued. Tubing Dimensions PEEK Tubing Color Volume 0.062 inch (1.575 mm) ID – 49.
Collect Using a Delay Function To run a Collect method using a delay function 1. On the Main screen, set the fraction size units (min, ml, or drops). 2. From the Advanced Collection screen, press Collect (F4). If collecting by volume, the delay volume can be entered directly. If collecting by time, the delay time is calculated from the delay volume divided by the flow rate. 3.
5 | Stand-Alone Operation Example: Collect All with Delay Figure 27 shows an example in which the delay is 0.2 min and the fraction size is 1 min. The diverter valve remains at divert during the delay time. The chart recorder makes the first tick mark (following the start/run mark) at 1.0 min, and each subsequent tick mark at 1 min intervals. The drophead advances at 1.2 min and continues advancing at 1 min intervals. Each fraction size is still 1 min. Fig. 27.
Collect Using a Delay Function Each subsequent tick mark occurs at 10-drop intervals. The drophead advances at 10-drop intervals following the start of the run. Note that if the delay size equals the fraction size, the start/run mark and the first event mark are superimposed. This means that the first observed tick mark (following the start/run mark) corresponds to the second fraction advance. Fig. 28. Example showing Collect All with a delay of 2 drops.
5 | Stand-Alone Operation At the start of the run an event mark is recorded. Initially the liquid is diverted to waste because the monitor signal is below threshold. When the UV signal rises above the specified threshold of 10%, an event mark is recorded and the volume corresponding to the delay time (or the delay volume) is collected in tube 1. At the end of this time, there is a tube advance and the peak is collected in 1-minute fractions.
Collect Using a Delay Function Example: Collection Using Time Windows and Delay Case 1, where start time = 5 min and delay time = 0 min Case 2, where start time = 5 min and delay time = 0.25 min Assume the following: Flow rate 1 ml/min Peak fraction size 1 min Time window #1 Start 5, End 8 Time window #2 Start 10, End 12 Note that each time or volume window is separated by a tube containing the delay volume.
5 | 74 Stand-Alone Operation | BioFrac Fraction Collector
6 LP/Econo Mode Operation In LP/Econo™ mode all aspects of fraction collection are controlled by an external controller. In this mode, only Rack, MultiRun, Start Tube, and End Tube parameters can be set from the fraction collector Main screen. Time and volume windows and threshold and delay functions are not programmable from the fraction collector in this mode.
6 | LP/Econo Mode Operation advances. If you are collecting in microplates or Titertube tubes choose a collection pattern: Serpentine, Row, or Column. Note: Whenever a new rack is selected, the start tube and end tube values are updated to reflect the maximum number of tubes available for the selected rack. 4. Choose the appropriate MultiRun mode for your experiment (see Table 6 on page 39 for a description of the MultiRun function). 5.
6. Press Engage (F1) to cause the fraction collector to move to the start tube, and listen for fraction advance and diverter valve signals. The BioFrac will wait for fraction advance commands from the external controller. Note: At any point during the run you can use the Run screen’s Stop function key (see The Run Screen in LP/Econo Mode on page 45) to stop the fraction collector.
6 | 78 LP/Econo Mode Operation | BioFrac Fraction Collector
7 Maintenance and Troubleshooting Maintenance The BioFrac™ fraction collector requires little maintenance to ensure reliable operation. To clean the case, first unplug the fraction collector. Use a damp cloth to wipe down the outer case. Avoid getting the power switch and rear panel connectors wet. Over time the clear glass ring inside the drophead may require cleaning. This ring protects the drop detector from splashes or dust buildup.
7 | Maintenance and Troubleshooting Fig. 31. Cleaning the drophead window. When finished using the BioFrac fraction collector, be sure to rinse all salts from the diverter valve with water. Leaving salt solution in the valve may cause salt crystals to form, which could plug or damage the valve. Rinsing salt from the valve will increase the valve’s lifetime.
Troubleshooting Troubleshooting Table 21 lists some common issues you may encounter using the BioFrac fraction collector, their causes, and possible solutions. Table 21. Issues, causes, and solutions Issue Possible Cause Solution No LCD display No power to unit. Check the power switch to be sure it is on. Check the power cord connections. Check the power at the outlet. Make sure the fraction collector display cable is plugged into the base unit. If problem persists, contact Bio-Rad.
7 | Maintenance and Troubleshooting Table 21. Issues, causes, and solutions, continued. Issue Possible Cause Solution Fraction collector causes a high backpressure Tubing is kinked. Replace tubing. Wrong tubing is being used. Connect the appropriate size tubing for the flow rate being used (see Chapter 3, System Configuration and Plumbing, on page 13). Rinse the diverter valve with water to remove salt crystals or other particulate matter. Divert valve or tubing is plugged.
A Bubble Filter Time The bubble filter time function is used in conjunction with the Peak Collection by Threshold. The bubble filter time function distinguishes true chromatographic peaks from unwanted signals such as electrical spikes or the passage of an air bubble. Such false signals are characterized by extremely fast rise and fall times. The bubble filter time function is entered at the end of the threshold programming sequence. The default value is off (bubble filter = 0 seconds).
A | Bubble Filter Time Fig. 32. Bubble filter time.
B Rear Panel Connector Information Fig. 33. 15-pin D, 8-pin mini-DIN, and 3-pin Combicon connectors.
B | Rear Panel Connector Information 15-Pin D Connector Table 22. 15-Pin D connector input signals Pin # Input Signals Type Active Wire Color* Description 1 ADVANCE Pulse Low Black Receives remote fraction advance signals (100 ms pulse). Active in LP/Econo™ mode. 5 LISTEN FOR REMOTE START Level Low Orange Connection of pin #5 and #15 (signal Ground) tells the fraction collector to wait for a remote start signal on pin #6. Active in Local mode.
15-Pin D Connector Table 23. 15-Pin D connector output signals Pin # Output Signals Type Active Wire Color Description 4 READY Level Low Green Indicates that the fraction collector diverter valve is connected and functioning properly. 9 START REMOTE Relay Closed Green/Black 10 DEVICE Relay used for starting a remote device such as a pump. Relay is closed when Run is pressed and opened when Stop is pressed. Active in Local mode.
B | Rear Panel Connector Information 8-Pin Mini-DIN Connector Table 24. 8-Pin mini DIN connector input signals Pin # Input Signals Type Active System Cable 7 Wire Color 2 WASTE/COLLECT Level Low Orange Description Controls diverter valve. Connection of pin #2 to pin #8 (signal ground) causes the diverter valve to change to collect. Controlled by a relay on a remote device. Active in LP/Econo mode. All input and output pins, except the relay terminals, are TTL compatible.
3-Pin Combicon Connector Table 25. 8-Pin mini DIN connector output signals, continued. Pin # Output Signals Type Active Wire Color Description 5 PEN UP/DOWN Level Low Brown Chart recorder control, pen up/down. Active in Local mode. 6, 7 ADS TERMINAL Level Low Purple blue Event marks for Method Start and Fraction Advance by way of a relay. Pins #6 and #7 are closed for 200 ms with each fraction advance. Active in Local mode. 8 SIGNAL GROUND Green Signal ground.
B 90 | Rear Panel Connector Information | BioFrac Fraction Collector
C Specifications The specifications for the BioFrac™ fraction collector follow in Table 27. Table 27. BioFrac fraction collector specifications Fractionation Time 0.01 to 9999.9 min Drop 1 to 99999 drops; flow rate 5.0 ml/min (maximum) Volume 0.02 to 99999 mls.
C | Specifications Table 27. BioFrac fraction collector specifications, continued. 3-way diverter valve 3-way diverter valve mounts on either the left or right column. Minimizes spillage during fraction advances. External operation Can be controlled by the BioLogic DuoFlow™ or BioLogic™ LP chromatography system, the Model EP-1 Econo™ pump, or other systems via external commands.
D Warranty and Ordering Information The BioFrac™ fraction collector is warranted for one year against defects in materials and workmanship. If any defects should occur during this warranty period, Bio-Rad Laboratories will replace the defective parts without charge.
D | Warranty and Ordering Information Warranty Information Model ___________________________________________________________________ Serial Number ____________________________________________________________ Date of Delivery ___________________________________________________________ Warranty Period ___________________________________________________________ Ordering Information Catalog Number Product Description 741-0002 BioFrac Fraction Collector, includes a 110 V power cord, rack F1(2), Econo™ syste
Ordering Information Catalog Number Product Description 731-8263 System Cable 3, mini-DIN to mini-DIN cable, for connecting the fraction collector to a BioLogic LP system controller or a Model EP-1 Econo pump (for control of the diverter valve) 731-8290 BioFrac Accessory Cable, 15-pin D to bare wires, for connecting the fraction collector to non–Bio-Rad equipment 731-8287 System Cable 16, 8-pin mini-DIN to DIN cable, for connecting the fraction collector to a Model 1327 chart recorder 731-8264 Sys
D | Warranty and Ordering Information 96 | BioFrac Fraction Collector
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