Horiba Scientific DeltaMyc J810018, Operation Manual

Page 1: ...D e l t a My c F l u o r e s c e n c eL i f e t i me I ma g i n gMi c r o s c o p eS y s t e m O p e r a t i o nMa n u a l P a r t n u mb e r J 8 1 0 0 1 8...

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Page 3: ...DeltaMyc Operation Manual J810018 7 Oct 2014 i DeltaMyc Microscope Fluorescence Lifetime Mapping System Operation Manual http www HORIBA com...

Page 4: ...form by any means including electronic or mechanical photocopying and recording without prior written permission from HORIBA Instruments Incorporated Requests for permission should be requested in wr...

Page 5: ...2 2 Environmental requirements 2 3 Electrical requirements 2 4 Unpacking and Installation 2 5 3 System Operation 3 1 Introduction 3 1 Controlling the hardware 3 3 Single point measurement 3 11 Lifeti...

Page 6: ...DeltaMyc Operation Manual J810018 7 Oct 2014 iv...

Page 7: ...ler PPD detector near IR up to 900 nm options available host computer with pre installed data acquisition cards and software DataStation measurement and control software DAS decay analysis software Fo...

Page 8: ...DeltaMyc 2 Requirements Installation Power and environmental requirements select the best spot for the instrument and set up 3 System Operation Operation of the fluorometer system and calibration in s...

Page 9: ...misuse of any such products including without limitation to persons injured directly or indirectly in connection with your use or operation of the products The foregoing indemnification shall in no e...

Page 10: ...from loss of use loss of data or loss of profits arising out of or in connection with our products or the use or possession there of HORIBA Instruments Incorporated is also in no event liable for dama...

Page 11: ...erstood and met HORIBA Instruments Incorporated is not responsi ble for damage arising out of improper use of the equipment A CAUTION notice denotes a hazard It calls at tention to an operating proced...

Page 12: ...precautions are not taken This symbol cautions the user that excessive hu midity if present can damage certain equipment Hot This symbol warns the user that hot equip ment may be present and could cre...

Page 13: ...uring your lifetime The long term risks for large cumulative exposure include premature aging of the skin wrinkles and most seriously skin cancer and cataract Damage to vision is likely following expo...

Page 14: ...this work NORMAL EYEGLASSES OR CONTACTS OFFER VERY LIMITED PROTECTION Training For the use of UV sources new users must be trained by another member of the labora tory who in the opinion of the member...

Page 15: ...r manent injury to the retina This type of injury can occur during a single prolonged ex posure Excessive exposure to visible light can result in skin and eye damage Visible light sources that are not...

Page 16: ...In addition the heat deposited in the cornea may be conducted to the lens of the eye This heating of the lens is believed to be the cause of so called glassblow ers cataracts because the heat transfe...

Page 17: ...es under the ob jectives for they can reflect or scatter the excitation light Also see safety notices relat ing to light sources Some DeltaDiode excitation sources generate laser light Do not look dir...

Page 18: ...DeltaMyc Operation Manual J810018 7 Oct 2014 Introduction 0 12...

Page 19: ...my of the DeltaMyc 1 1 1 Anatomy of the DeltaMyc Introduction The DeltaMyc is made up of a large number of devices and hardware Therefore this chapter is designed to help you understand how all the de...

Page 20: ...DeltaMyc Operation Manual J810018 7 Oct 2014 Anatomy of the DeltaMyc 1 2 Block diagram Following is a block diagram showing how the major sections of the DeltaMyc are inter connected...

Page 21: ...natomy of the DeltaMyc 1 3 DeltaMyc Inverted IX73 microscope Tungsten halogen lamp x y Stage PPD detector PPD detec tor power supply DeltaDiode controller TCSPC electronics Joystick Stage controller D...

Page 22: ...natomy of the DeltaMyc 1 4 DeltaMyc Upright BX53 microscope Tungsten halogen lamp x y Stage PPD detector PPD detec tor power supply DeltaDiode controller TCSPC electronics Stage controller Joystick De...

Page 23: ...training includes but is not limited to Understanding the risks of exposure to ultraviolet visible and infrared light and how to avoid unsafe exposures to these types of radiation Handling various lig...

Page 24: ...ts Installation 2 2 Surface requirements A sturdy table or bench top Surface must hold 90 kg 200 lbs Surface should be about 27 72 69 cm 183 cm to hold spectrofluorometer com puter and accessories com...

Page 25: ...ation Caution Excessive humidity can damage the optics Caution Airflow inside the FluoroHub module de pends on the top cover being in place Ensure ad equate airflow to avoid damage to certain compo ne...

Page 26: ...djacent to the IEC mains power inlet Fuse ratings are listed in Specifications Note HORIBA Scientific recom mends connecting the host com puter monitor and printer to a sin gle surge protector to make...

Page 27: ...ed as close as possible to the selected location DeltaMyc carton contents Quantity Item Part number 1 Microscope frame BX73 or IX73 with one port for the at tachment of the microscope lamp in reflecti...

Page 28: ...pplicable 1 DeltaHub 1 Host computer with data acquisition cards if applicable 1 or more DeltaDiode light source s specified in the order 1 DeltaDiode controller if applicable 1 PPD photon detector Al...

Page 29: ...ace the microscope frame on a flat surface Refer to the microscope instructions to set up the microscope lamp and illuminator if they are not already attached to the microscope frame Place the DeltaHu...

Page 30: ...witched on and off using the switch next to the inlet The interlock plug must be in place to ensure proper operation Turn ON OFF the laser power using the key on the front panel The DeltaDiode head co...

Page 31: ...d on be cause the various USB devices are not necessarily connected to the same USB connector they were previously using For further details please refer to the application user guides for DataStation...

Page 32: ...y of connections Type of connec tion Number of connections Equipment AC mains 110 V or 220 V 7 Microscope frame DeltaHub PC Monitor Motorized stage controller Camera DC adaptor DeltaDiode controller U...

Page 33: ...f the decay time either from a previous measurement or lit erature Starting the software 1 After checking that the appropriate hardware is connected turn on the host computer Delta Hub the Tango stage...

Page 34: ...ty can be varied You can select a suitable filter cube in the illuminator and use the binoculars top camera to view image the sample The laser is only on when the camera only position Note that in th...

Page 35: ...f the microscope motorized stage and the camera pixels and lets the system know where the pinhole is located in the horizontal plane Therefore run the calibration routine each time that the system con...

Page 36: ...stage camera relative position The Microscope Calibration window is shown in figure 13 and the calibration proce dure is detailed below Fig 13 Microscope Calibration window For this step the motorized...

Page 37: ...hest magnification objective Calibration of laser reference position and diameter if applicable The calibration procedure is detailed below Using the internal camera the laser position is identified o...

Page 38: ...a on the microscope stage c Rotate the filter cube dial to the filter cube that matches your DeltaDiode d Make sure the dual lamp adapter selec tor knob is upward so that the light guide illuminates t...

Page 39: ...Myc Operation Manual J810018 7 Oct 2014 Operation 3 7 f Click the TCSPC Single point icon then click the New button The main window appears g Click Stage calibration The Calibration warning window app...

Page 40: ...ia gain control j Select pinhole on the camera adapter 2 Calibrate the instrument a Shut off the white light to the micro scope by pulling down on the dual lamp adapter selector knob b Gently remove t...

Page 41: ...DeltaMyc Operation Manual J810018 7 Oct 2014 Operation 3 9 d Illuminate the pinhole with a small flashlight You should see the circular image of the pinhole on the monitor...

Page 42: ...tate the knob to match the blue circle to the size of the image f Remove the flashlight return the filter and reinstall the detector tight ening the set screw g Redo stage calibration h In the main wi...

Page 43: ...tton then click the OK button k First change the calibration distance to 0 05 mm l Identify a particular site on the sample and click The image shifts to center upon that site m Re identify that same...

Page 44: ...ation Manual J810018 7 Oct 2014 Operation 3 12 c Pull down the dual lamp adapter selector knob so that the DeltaDiode illuminates the microscope The image of the sample s fluorescence should appear on...

Page 45: ...al J810018 7 Oct 2014 Operation 3 13 d Move the DeltaDiode s alignment adapter knobs to center the laser illu mination on the crosshair target 4 Optional Adjust software position by clicking Adjust so...

Page 46: ...n be accessed manually to read the current position and move the stage to a new location The Advanced button gives access Fig 16 to advanced options for the stage In this window you can select the typ...

Page 47: ...first In the measurement tree four items are now available This tree is illustrated in Fig 18 Figure 18 Measurement tree for microscope single point measurements The Photos folder contains all the pic...

Page 48: ...elect position window opens Fig 19 Fig 19 Select position window e Adjust the camera settings using the Video settings button and Video format button Both of these buttons open different Properties wi...

Page 49: ...ove the sample click on the image on the feature of interest The point of interest is then moved at the pinhole location and becomes ready for the measurement i Save the live image to a BMP file using...

Page 50: ...lter beam splitter position Data display 1 Click on the Prompt and Decay items to display the measurement data as in standard TCSPC mode 2 To display any pictures that have been added to the DAS file...

Page 51: ...DeltaMyc Operation Manual J810018 7 Oct 2014 Operation 3 19 Open the DAS files with the DAS6 software for further analysis...

Page 52: ...It is possible to map non contiguous non rectangular areas TCSPC FLIM Acquisition time ranges from 5 ms to 1 s per decay Only rectan gular areas can be mapped Anchor control An additional anchor contr...

Page 53: ...nchor control The Select position window appears Fig 21 The purpose of this window is to store the location and the picture of the area of the sample that is to be mapped Fig 21 Select position window...

Page 54: ...parameters and the measurement locations on the anchored sample are con figured using the microscope scan wizard 1 Click the Click here to begin item in the meas urement tree The wizard window is disp...

Page 55: ...e delay between the prompt and the decay curve The logarithm of the decay data is then fitted with this calculated lifetime in its linear part If no prompt was acquired the estimated lifetime is calcu...

Page 56: ...se points radio button b Click on the center of the cell to be deleted or click and drag to draw over an area of points to be removed The resolution of the grid is defined by the laser spot diameter H...

Page 57: ...nue The data are now ready for review If a prompt is measured after the decays the data analysis will occur after this last ac quisition Therefore it may take a few minutes before this dialog box appe...

Page 58: ...layed To obtain more information about a particular measurement location move the mouse pointer to this location The relevant data are shown in the Information section To display the data curve of a p...

Page 59: ...the first zoom area corner drag and click to define the second cor ner of the zoom area The zoom area is then displayed in a new window Utilities Print map Offers various options for printing the map...

Page 60: ...2D Microscope Map window To display the data of a particular decay click on the relevant Decays item The data can also be ex ported using the functions on the item s popup menu Starting a new Microsc...

Page 61: ...dard TCSPC measurement For a full de scription of the software we recommend that you consult the DAS6 manual 1 Click the DAS6 icon on the desktop to start the DAS6 software 2 Open the relevant microsc...

Page 62: ...obal sum fit For more information regarding these algorithms see the DAS6 manual 6 Choose the Fit button which opens the 1 5 exponentials window Fig 29 1 5 exponentials window a Select the desired num...

Page 63: ...hted residuals indicates how the fitted function differs from the data and a reduced 2 value CHISQ or XSQ Other indicators are also provided The usual values of interest are the lifetime T value its e...

Page 64: ...ticular pixel is displayed across the top of the screen when hovering over the pixels with the mouse A histogram displays the statistical distribution of the fit parameter across the sample The displa...

Page 65: ...ting 4 1 4 Maintenance Troubleshoot ing Maintenance If the instrument requires cleaning then disconnect it from the AC mains supply Ap ply a mild detergent solution using a soft slightly damp cloth Al...

Page 66: ...pe frame and the wall socket The bulb needs to be replaced Please check the Olympus documenta tion supplied with the microscope frame and replace the lamp bulb The lamp is ON but the intensity is set...

Page 67: ...h the DeltaHub With the Lifetime module With the Tango stage controller The hardware is not ON Switch the DeltaHub ON Switch the Tango controller ON Connect the power supply unit cable to the lifetime...

Page 68: ...tage have been moved or replaced Run the calibration procedure If there is no move ment at all the stage may be not operation al Make sure that the Tango control ler is ON and check the XY ca bles The...

Page 69: ...fic support in tracing the problem The most recent log file is always named APP LOG where APP is the name of the application that produced the file i e DataStation or DAS6 The log files are al ways cr...

Page 70: ...back on 2 Make sure all accessories are properly config ured and turned on as needed 3 Run the verification tests to make sure the sys tem is properly calibrated Print the spectrum or table for each...

Page 71: ...ifferent port you may have to reinstall the appropriate USB driver Usually this process is handled by Windows In some cases you may have to point to the driver manually All drivers are supplied on CDs...

Page 72: ...DeltaMyc Operation Manual J810018 7 Oct 2014 USB Port Configuration 5 2...

Page 73: ...of the DeltaMyc spectrometer fol low Microscope Microscope type Based on upright Olympus IX53 or IX73 Objectives Plan achromat 10 and 60 other magnifications available Turret with five positions avai...

Page 74: ...it spread time 200 ps Dark count 80 cps Filters Available positions Eight manually selected interchangeable cubes Excitation filters Optional 10 nm bandpass filters or short pass filters Dichroic beam...

Page 75: ...itter 3 22 4 2 BF 4 2 bitmap file 3 22 block diagram 1 2 BMP file 3 17 BS 4 2 BX73 2 5 C C mount 2 5 cables 2 4 7 4 3 4 calibration procedure 3 3 5 3 15 3 17 Calibration warning window 3 7 caution not...

Page 76: ...xcitation wavelength 3 1 Expansion port A 2 8 9 Export data 3 27 F face shield 0 6 File menu 3 18 3 20 3 28 filters 6 2 filter cube 3 2 3 3 6 4 2 4 4 Fit button 3 30 Fit button 3 30 FLIM icon 3 28 flu...

Page 77: ...ocket 2 8 power supply 3 1 PPD detector 0 1 2 6 2 8 4 4 6 1 2 Print map 3 27 Prompt item 3 15 3 18 3 28 Properties window 3 16 protective gloves 0 6 R Ratemeters window 3 18 Read this manual notice 0...

Page 78: ...e Video settings button 3 16 Time resolved mode 3 2 Tools 3 27 Transparency slider 3 26 trinocular 2 5 4 4 troubleshooting 4 1 2 turret 6 1 U ultraviolet light notice 0 5 unpacking and installation 2...

Page 79: ...up application images are collaged in the overall design Beginning from a nano size element the scale of the story develops all the way to the Earth with a gentle flow of the water 3880 Park Avenue Ed...

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