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Prior PureFocus850 User Manual

Laser autofocus system
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More to PRIOR than meets the eye
PureFocus850
User manual
Version issue No. - 1.7
For more information visit www.prior.com

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Summary of Contents for Prior PureFocus850

  • Page 1 More to PRIOR than meets the eye PureFocus850 User manual Version issue No. - 1.7 For more information visit www.prior.com...
  • Page 2 Thank you for purchasing this product from Prior Scientific – we’re confident it will be a reliable and useful addition to your microscopy system. Please take the time to read and understand this manual before using this product – it contains not only important operating instructions but also vital safety information.

  • Page 3: Table Of Contents

    More to PRIOR than meets the eye Contents Section 1 Safety Information Section 2 Product description Section 3 Unpacking the system Section 4 Introduction Working principle Sample types Types of microscope system Section 5 Installation Installing the head Initalizing the head...

  • Page 4: Safety Information

    PureFocus850. If you have any questions do not hesitate to contact Prior Scientific. If you intend to use this unit in a manner not specified by Prior in this manual, contact Prior beforehand.

  • Page 5: Product Description

    Section 2 Product description The Prior Scientific PureFocus850 is an advanced, integrated, unit comprising of an IR laser diode, precision optical components, detector and signal processing electronics with on-board micro controller. The system allows optimum visual focus to be found and maintained on a microscope system for a range of different sample types, microscope objectives and imaging methods.

  • Page 6: Unpacking The System

    Section 3 Unpacking the system This guide is an introduction to installing a Prior PureFocus850. Please note that instructions for specific microscopes and parts may vary. Consult Prior Scientific if you are unsure. PF850 PureFocus Controller, Head, Cables & PSU...
  • Page 7 More to PRIOR than meets the eye 1. PF system In your shipment you should recieve: PF head PF controller Connecting cable PF controller PF head Power supply USB cable Stepper motor (optional) Piezo BNC cable (optional) Dovetail kit (optional)

  • Page 8: Introduction

    Section 4 Introduction 4.1 Working principle The Pure Focus operates using a sensor with mutiple pixels. Half the aperture of a collimated laser beam is blocked via a knife edge and directed into the back of the microscope objective. The laser light is focussed to a line on a reflective surface at the sample and then reflected back through the objective and directed towards the sensor forming a corresponding line on the sensor at the centre point.
  • Page 9 ERROR = TARGET - POS = TARGET - (A-B)/(A+B) This signal is fed into a PID controller and the output of this controller is sent to either the Prior stepper motor or to an analogue voltage for driving an external piezo controller.

  • Page 10: Sample Types

    4.2 Sample types PureFocus works with a variety of different samples: Permanently mounted slides These are specimens that are fixed in a mounting media usually with refractive index matched to the glass coverslip (n≈1.5). When working with dry objectives with such samples a reflective signal is only available from the outside of the coverslip.

  • Page 11: Types Of Microscope System

    More to PRIOR than meets the eye 4.3 Types of microscope system PureFocus works on a range of different microscopes which use infinity corrected optics. The PureFocus head sits in the infinity space of the optical path between the objective and tube lens.

  • Page 12: Installation

    5.1 Installing the head Install the microscope dovetail kit onto the PureFocus head unit. For information on installation for inverted microscopes please contact a Prior representative. Mount the PureFocus head unit onto the microscope between the objectives and tube lens, in the inifinity space (there are multiple places it could be mounted in the infinity space depending on the microscope).

  • Page 13: Initalizing The Head

    Turn Allen screw labelled Blade (4) clockwise to the limit to remove knife edge from beam path Turn on the control box and run the PureFocus850.exe Graphical User Interface (GUI)* *Please note you will have to download the GUI from the prior.com website.
  • Page 14 13. Set Offset lens back to default position (Offset>Goto Factory Home) 14. Using your camera imaging software and PureFocus controller nob, or microscope manual *camera.exe can be downloaded from the prior.com website along with the GUI software Version issue No. - 1.7...
  • Page 15 More to PRIOR than meets the eye adjustment to focus on the mirror. 15. Set the laser power to 10 (Laser > Set Power , value 10) 16. In the main GUI, go into setup mode (Setup) and observe the graph 17.

  • Page 16: Controller Motor Connections

    ‘Do you wish to apply to all objectives?’ click Yes. The background is now applied to all objectives 24. Click save to flash (Settings > Save to Flash) 25. Close the PureFocus850.exe GUI 26. The unit should now be ready to use 5.3 Controller Motor Connections The 15 way D connector on the Pure Focus controller allows connection to either a stepper motor or a piezo actuator system which can accept an analogue 0-10V input.

  • Page 17: Setting Up For Stepper Focus Control

    BNC connector to the desired piezo controller, this BNC outputs 0-10V and operates nominally centred at 5V Turn on the piezo controller and then the PureFocus control box, run PureFocus850.exe GUI Move to a long working distance objective, such as 4x Change mode to piezo mode (Mode >...

  • Page 18: Controller Keypad And Digipot

    Section 6 6.1 Controller keypad and digipot Digipot Turning adjusts either the Z position or the offset depending on the mode selected (See Offset/Focus). If the servo is active then Z position control becomes automated by the PureFocus and the digipot can no longer adjust the Z position.

  • Page 19: Inital Setup And Operation

    More to PRIOR than meets the eye This table describes each indictor: OBJ1, OBJ2, OBJ3, OBJ4, Displays the current working objective, current parameters* are OBJ5, OBJ6 associated with this objective Appears when no sample is detected (see SAMPLE flag) Appears when sample is in focus (see FOCUS flag)
  • Page 20 The error signal can be seen visually and numerically on the GUI main screen. If these methods fail please contact Prior Scientific with details of the problem, the sample or objective may be incompatible with PureFocus.
  • Page 21 More to PRIOR than meets the eye objectives there can be a strong back reflection from the optics and running at lower laser powers is preferable, although focus capture range and speed will be reduced. 6.3.5 Activating the focus servo It should now be possible to activate the servo and see if PureFocus is able to hold focus.

  • Page 22: Routine Operation With The Gui

    6.4 Routine operation with the GUI Please note, the following is written in relation to the GUI ‘PureFocus.exe’ however the majority of the functionality discussed in this section can be carried out directly using ASCII commands, allowing integration into custom software.
  • Page 23 More to PRIOR than meets the eye The pre configured parameters set the threshold levels for the above flags to sensible starting levels. It may be required to adjust these levels to perform optimally for your application; this can be done via the `Flags’...
  • Page 24 menu (Mode > Focus Direction Inverted), which also inverts the digipot direction. The maximum speed and acceleration of the Z-axis can also be changed via the `Z-Speed’ dialogue box. For troubleshooting or assessing the behaviour of the error signal, simple Z scans can be carried out using the `Z-Scan’...
  • Page 25 More to PRIOR than meets the eye 6.4.8 Servo control Pressing `Servo on’ and `Servo off’ on the main GUI screen turns focus servo control on and off respectively in the same manner as the `Servo on/off’ button on the keypad. When Servo control is on the focus correction is shown in real time on the GUI main screen.
  • Page 26 6.4.12 Setup mode Setup mode gives the ability to live-view the PureFocus sensor directly, running at a significantly lower update rate. Although all parameters are still updated in real time on the main GUI screen and dialogue boxes, the servo cannot be activated in this mode due to the lower update rate. After Installation and Initialization, Setup mode is not required for the typical user, it is only required if one wishes to do a full manual setup for a particular objective.

  • Page 27: Ascii Command Set

    More to PRIOR than meets the eye Section 7 ASCII Command set PureFocus communicates with a Baud rate of 460800, Prior Terminal can be downloaded from the Prior website. Signal setting commands COMMAND RESPONSE DESCRIPTION PINHOLE Centre, width Returns pinhole centre and width...
  • Page 28 Servo setting commands SERVO Queries current servo state, 0=off, 1=on SERVO,N Turns servo on or off respectively,0=off, 1=on Query proportional gain for current objective, f is integer KP,F Sets proportional gain value for current objective, f is integer Query integral gain for current objective, f is integer KI,F Sets integral gain value for current...
  • Page 29 More to PRIOR than meets the eye Flags and flag setting commands SAMPLE Returns current sample state: s=0, sample not detected s=1, sample detected s=1 when A+B>SAMPLEL SAMPLEL Returns the low threshold l for determining the sample state for the...
  • Page 30 INTERFACEH Returns the high threshold h for determining the interface state for the current objective INTERFACEH,h sets the high threshold h for determining the interface state for the current objective INTERFACEL returns the low threshold l for determining the interface state for the current objective INTERFACEL,l sets the low threshold l for determining...
  • Page 31 More to PRIOR than meets the eye LIST 0 Example: Lists all parameters that are set for the current objective OBJ,1 KP,2000.00 KD,0.00 KI,0.00 LASER,0 TARGET, 0 FOCUSL,2000 FOCUSH,4095 FOCUSR,0.02 INTERFACEH,0 INTERFACEL,0 OUT- LIM,-100000,100000 SAMPLEL,50000 ABON,0 PINHOLE,750,750 REGION,D,0,0 BACK- GROUND,0,0,0,0...
  • Page 32 Offset lens commands LENSH Sends the offset lens to its home position LENSV,v Moves offset lens at velocity v steps/s LENSP Returns current offset lens position in steps (there are 25600 steps/mm) LENS$ Returns offset lens motion status: n=0, idle n=1, moving LENSACC,a Set acceleration of offset lens...
  • Page 33 More to PRIOR than meets the eye Move focus up by C steps Move focus down C steps Returns default step size for U and D commands units of 100 nm Sets default step size for U and D commands units of 100 nm...
  • Page 34 PIEZO Returns current raw piezo DAC output when in piezo mode (see above) System commands DATE Prior Scientific Instru- Returns date of software compilation ments OptiScan LF and build number, (n) Version (n) complied (date & time) SERIAL decimal number...
  • Page 35 More to PRIOR than meets the eye REGION,r,s,f REGION,r,s,f Sets region r (r = A,B,C or D) start and finish pixel to use on line sensor for computing A, B, C and D s,f = integers in range 0 to 1499...

  • Page 36: Replacement, Repairs, Returns And Spare Parts

    Prior Scientific directly. If you wish, or are advised to, return your unit, a RMA number must first be obtained by filling in a returns form located on the prior.com website. We will then send you an RMA number which should be added to a decontamination form again located on the Prior Scientific website.

  • Page 37: Troubleshooting

    More to PRIOR than meets the eye Section 9 Troubleshooting Problem: The back reflection level is too high and/or the signal appears to have unstable noise components that “bounce” up and down. Solution: Back reflected laser light is returning directly to the sensor and/or laser. Attempt to deliberately misalign the dichroic in one axis by making very small adjustments to Dichroic adjustment 2 (Nominal 45°), see page 11.

  • Page 38: Appendices

    Appendices Recently the PF850 controller has had an upgrade/change to the USB connection hardware. The company FTDI Ltd. manufactures the hardware and constantly updates the associated Windows USB Driver. The new USB hardware and interface driver is Windows Hardware Quality Labs (WHQL) tested and will automatically download the latest drivers when connected to a PC with internet access.
  • Page 39 More to PRIOR than meets the eye In the Properties window, click on the Driver tab and view the Driver Date: If it is older than 2016 or if the Driver Provider is not listed as FTDI, see the image below, a new driver needs to be installed. If the PC does not have internet access the files can be loaded onto the PC via a USB Stick It is recommended to uninstall the existing FTDI drivers.
  • Page 40 Extract all the files and run the CDMuninstallaerGUI. Click the Add button on the right and the Vendor ID and Product ID will be populated inside the blank area. Download the latest USB Driver version from: http://www.ftdichip.com/Drivers/VCP.htm Download the proper 32 or 64 bit driver as shown below. Extract all of the files to the desktop or other download folder.
  • Page 41 More to PRIOR than meets the eye Browse to the location of the saved Driver and click Next Windows will then update the Driver using the downloaded files. A successful installation will show a new date in the USB properties as shown below.
  • Page 42 Software”, as in some instances Windows will not assign a proper COM port the first time the driver is updated. At this point it is advisable to adjust the USB latency timer from the standard 16ms down to 2ms. Right click on the new COM port and choose Properties.
  • Page 43 More to PRIOR than meets the eye Click OK twice to save the Latency Timer change. For more information visit www.prior.com...
  • Page 44 Version issue No. - 1.7...
  • Page 45 More to PRIOR than meets the eye For more information visit www.prior.com...
  • Page 46 More to PRIOR than meets the eye PureFocus850 User manual WORLDWIDE DISTRIBUTION Prior Scientific Ltd Prior Scientific Inc Prior Scientific GmbH Prior Scientific KK Prior Scientific China Cambridge, UK Rockland MA U.S.A Jena, Germany Tokyo, Japan Suzhou, China t: +44 (0)1223 881711...

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