フローチャート式 MetaFluor簡易取扱説明書
Contents
1. Ratio Molerular Devces apan KK ImggngTeam 415 4 Command bar New Cfg Acd Cfg Exp Events Focus Set Time Lapse Set Disp Region Ratio References Cfg Graphs cfg Graph Configure Graph nrfiEure Graphs pd heasured walue ppears on Fahh Wawueleneth 1 eraEe Intensity Graph 1 Wawveleneth 2 werage Intensity Graph12. MetaFluor EyEnt Marks FE hark FE Mgxt II Lielete hawe Clear Lnad Pinuhla ClicE to mark ewent Pra Fa tatus Beep on Ewent Less lt Mark Gurrent Ewent Fh Manwe tn Mext Ewent FE HH TE hift F HH DFP hft F Settines tor hew Ewent add hE Shiftt F3 Ewent Text Trigeger out Shift F4 Dount domn Timer nD a seconds no timerl Elect Er rnalto be executed when Event is marked ho inurn is salectad1 hark Mem Ewent No dd Ne Ewent to List Inse Event Marks Mark New Event Now Inf Enable Timer ly darkewent on messaEes om Clampex Setting for New Event Event Text
3. Graph XX Y X Graph Setting ranh Settinegs Samples raph Traces 5 TS Leeend E aranCe Dlick button to chanee the way the Eraph looks Cinlor scheme 1 1 Molecular Devoes apan KK Imggng Team 5 15 Command bar New Cfg Acq Cfg Exp Cfg Graphs Focus Set Time Lapse Region Ratio Set Disp References Events Run Experiment Event Marks
4. _a Waveleneth 1 100 EL SSKDDD Lm po co Intensity 190 20D Time Seconds File DDELogtExcel Book1 Sheet1 11 Date Tue Jun 26 2007 16 27 57 795 0 Wawvelength Names Wavelength 1 Wawelength 2Wavelength 3wawelength 4Wawelength 5 Time sec 66 65 1 119 27 122 27 125 27 128 26 131 26 Region 1 Lncatinn 87 29 Size 38 44 Area 1292 Region 2 Location 183 43 Size 40 30 Area 940 Region 3 Location 153 9 Size 29 25 Area 561 Region 4 Location 68 128 Size 30 33 Area 770 Time sec R1 R1 R2 R1 R3 R1 R4 R1 0 Clock reset to 00 1 04 1 23 1 11 1 14 4 38 1 23 1 11 1 15 7 48 1 23 1 11 1 14 10 48 1 24 1 12 1 15 1 22 1 11 1 13 Region Excel text CO J IT rw PO Molecular Devoes apan KK Imggng Teagm 131 Command bar New Cfg Acq Cfg Exp Cfg Graphs Events Focus
5. inf inf 3 es ExnerimEnt nntrnl PF tatus W1 W2 LogData File nat npen ave Images File nat npen Ratio1 lp Savs Fatins R TI wcle et Timelapse FS ETn Clock Fncus Case Save Ratios Ratio Ratio TIF TIF Log Data Save Ratio Save Images Molecular Devces apan KK Imggng Tesam 815 8 Command bar New Cfg Acd
6. Set Time Lapse Set Disp Region Ratio References Command Bar Close Close MetaFluor Load HetaFluor Tnu hawe chnseh tn close the experiment Fress lnse experitehnt ahd sawe proatoacol Cilose experiment but dnnt sawe protocal CahoEL Hannt close the experiment gay Prntnrnl escription FF YFF FRET Timelapse Interwval 3
7. INF O Load Experiment Control Panel Event List We Event Le EK i Pr Experiment Fontrol hh lee Ewents om this eperiment tatus lack reset to Loe Data File nnt npen Addad lanamwein Sawe lmases Fils not open addad E T Sawe Ratios Fils nnt open SE Frame 1 1 515 Imaga Time Oh5 Seconds _FS Reverse F4 Forward Clpse im Rd close Experiment Control Panel Hrame Image Time F3 Forward FE3 Reverse F2 Pause Load Retio
8. Regions Configure Experiment Cfg Exp 3 Ave Sum Area Measurements Dynamic Data Exchange DDE Microsoft Excel A Text File Log 2 Experinent Control P Save Image Re EE Lapats IFWenetanenl sn W1 W2 Logg Pata File nat npan Iw Sawe lmages FLLIHRD IMF Save Ratios File nnt open Cycle Set Timelapse PS in Clock Focus Clnse MetaFluor INEF inf 001
9. nnfiEurE EnuIgitinr Wauelsngth to nTEUFE ET uelength hame MT se Selscted Wawveleneth as hame m Linstnmi Defned WamalsnEth 5 WW malahEth hanEinE Hardware lllumin atinn Cre cg IFRET YFP 2 Wauweleneth Use Waweheneth 1 js the numerator for Ratin 1 Ratin 1 WR Fnsitinhi SetzZ Position for this Wawelength to Ciameras Einnineg and Sub RieeEion LiEital Ciamers cquisition Parameters Laft Didth E Exposure Time OO Mitiseconds lt Top Heieht 512 an any 5 gnnne EE ImaEe SIE 1 he TransfEF Speetd lio Hz EM Gainy lt elect Fiegion Camera Shutter Aways Open lt se Entire Image EMGain I Insg Wavelength1 MetaFluor Ratio1 Wavelength1 Wavelength2 ex W1 fura2 340nm W2 fura2 380nm Ratio2 Wavelength4 Wavelength5 ex W4 FRET CFP W5 FRET YFP Wavelength3 DIC Fluo3 Custom Defined Wavelength Wavelength
10. Add New Event to List F5 Mark Run Experiment Event Mark Hotkeys Granhs Ltilities Calibration Jdournals Windows Analog Experiment Cantral Panel Stop Acquisitionl F2 One Acquisition F3 Resume Acquisition F4 ero Clock CtriZ Save Last Acquired Images F7 Scale 16 bit Images Raference Images Select Save Region Eyent Marks Event Mark Hatkeys Mark Current Event F5 howe to Next Event F65 ei Add ATP Shift F1 RW Add ADP Shift F2 Bcdure Stream Add MP Shift F3 hove All Regions Trlgger out Shift F4 lign Wiayelengths Edit Conditions Molecular Deics apan KK Imggng Team 615 6 Command bar New Cfg Acd Cfg Exp Cfg Graphs Events Set Time Lapse
11. Histogram of Image Store for all Wave Configure Acquisition 2 Use Wave Settings Configure Acquisition 22 Allow items to differ for each wavelength Illumination Use Wave Settings Tul1 Chip Store as Acq Rgn Live Z
12. Region Region Acquire Images 1 OK Edit Region Tools Region Tool Region Tools Region Region Cfg Graphs Events Focus DC Set Time Lapse Region Tools Set Disp ITF References 2 Clear DReset Fnlors Dreate Reginn round bjects hin Size 1 kak Sige 1 ODO EE ar fF ofreEinns 55 1 NReeions Defned 3 ctive Reeion 1 weraEes Intensity 2512 53 hortcuts 6pane Region Save Load Region Region Region R
13. 7 Region 10 F3 Forward PEvent List Molecular Devoes apan KK Imggng Tesam 15 15
14. Pause OClose Molecular Devces apan KK Imggng Team 915 Command bar New Cfg Acd Cfg Exp Cfg Graphs Events Focus Set Time Lapse Region Ratio References Set Disp Command Bar Set Disp WImaee Disnlay Controls Waveleneth 1 75 Wavelength 1 75 lmiaga Window Properties Cisplaw IMan achrome Brightnsss dn Ciontrast 1 Thrashnld state mr f Inalusiue Esimlusiue Wavelength1 Brightness Contrast Display Monochrome Pseudocolor
15. Ns HefErEriEE ImaEE aeleneth Farameter ckEF ur Farameter hadine INans na Mene na one na None na 5 ipad ns Wavelength Background Shaging Image Load Acquire ho imags 1 Wama laEnEth 4 ae lEnETth 5 orrect hadine Region Parameter 10 Region Region Region Region Subtract Backgrounds Correct Shading Region 1 Parameter Region
16. Area Regions 9 Experiment Control Panel Save Image Ratio 1 2 Save Ratio Update 1 ON O Wavelength Wavelength 1 FRET YFP Wavelength 2 FRET CFP Wavelength 1 FRET YFP Wavelength 2 FRET CFP Ratio 1 W1 W2 2 MetaFluor 1Image Wavelength 1 Wavelength 2 Ratio 1 2 Ratio
17. llumination Wavelength Z MetaFluor Binning Sub Region Select Region Use Entire Image Rxposure Time Gain Wavelength Allow items to differ for each wavelength Digital Camera Acqguisition Parameters Wavelength 6 Focus Molecular Devoes apan KK Imggng Team 15 Command bar
18. Set Disp Region Ratio References Focus Focus Focus XY Focus Command bar Focus Control Experiment Control Panel ImaEa Se 1 Hg Ex omo ms Eh Gain ain Gain 3 5 1 nHz Eh Use dc Resion Store as cd Rgrl 4 GTR pk Full Chip Store tor all WWawgs lllu min ation Waweleneth 3 Use Wawe Sattings tore tor this Wawe uelEnEth 4 Waweleneth 5 Toeele Shutter ly pen on Start Focus lose ter MAUIFe 15354 327BB dd152 55535 Histngrarm of Image uton Scaline Start F 5 F LE Lal hn toscale Hi _Use Wave Settings Gtore tor this Wawe ilpse Position 1 Foto se WWawe Gtnre far this Mawe Start Focusing Live Live
19. Cfg Exp R Cfg Graphs Events Focus Set Disp Region Ratio References Set Timelapse Experiment Control Panel Set Timelapse Set Pe Timelapse Intervyal Number of Acquisitions Experiment nntrnl P IE3 tatus Logg Data Sawe lmases Fils nat npen File not open Sawe Ratios File not CFS Fncus Insg Set Timelapse Het Tmelapse E Seconds Timelapse Interwal lumber of Acquisitions 1 1 Timelapse Interval rm Interval X wir of Acquisitions Number of Acquisitions
20. Subtract Background Wavelength 1 2 Region Moecular Deics apan KK Imggng Team 121 Command bar New Cfg Acd Cfg Exp Cfg Graphs 8 Events Focus Set Time Lapse Set Disp Region Ratio References Experiment Control Panel see Experiment Control P tatu Ip Loe Lata LE Sawe Images FLU RIMF _ 7 Save Ratios RATIO wcla Set Timelapse 6 Facus Close F4 Acquire F2 Pause F4 Acquire Zero ClocE
21. New Cfg Acd Cfg Graphs Events Focus Set Time Lapse Set Disp Region Ratio References Cfg Exp Configure Experiment nrfigure Experiment PTain ll pdate WaelenEth 4 n Of Waweleneth 5 n ff Ratio 1 Dn Of Fiatin g On Of Halcium ly Fatio CT WE ly Update Fatia WE Palibrated Ratio 1 ancgl 1 2 2 1 Experiment Control Panel 7 Log Data Avg Sum
22. 4 0 Goto 0 Store for this Wave Configure Acquisition 2 Molecular Devoes apan KK Imggng Tegm 715 7 Command bar New Cfg Acd Cfg Exp Cfg Graphs Events Focus Set Time Lapse Set Disp Region Ratio References 1 Experiment CnntrnlF tatus we LgE Data DiDE Sawe lrmiage File nat open ave Ratins File nat open Test File Pycls et Timelapse PS Fn Clock Fncus lnsg Cancel Rxperiment Control Panel Log Data
23. Wawveleneth 3 weraEe Intensity one Wawueleneth eraEe Intensity Ione Wawelenegth 5 verges Intensity one Ratin 1 waraga Ratio IEraphg Ratin amp weraEe Ratio None Calibrated walue ionfeEure Graph is Graph Graph 1 Title I raph 1 Show Graph mt la one FeEI Hi Must be detfned hefmre aEFahh can be displayed Time Seconds RaneEe 5nn Ticks 1 Tick marks ewery BH HH et a ThE Wa al Measured Value Appears on Graph Line Type Wavelength1 Solid Wavelength2 Dotted Graph1 Ratio1 Solid Title lIntensity Range tn EEgan Ticks Tick marks ewvery 5BHH HI In na Ft LnE calinEg Graph2 Graph 1 2 3 4 5 Show Graph Appears on Graph Graph
24. F et ID iTiaEa Cc uisitinn Ct Exp Choose the images to acquire Scale 15 Scale 15 hit imaees RetererncesiReference subtraction hadinE1irmaEe et Disp TErightness contrast and thresholds fg GraphslDontfeure measurement eraphs Command Bar More Less Command Bar Molecular Devoes apan KK Imggng Team 2 15 2 Command bar New Cfg Exp Cfg Graphs Events Focus Set Time Lapse Set Disp Region Ratio References Cfg Acq Configure Acquisition Tlumination Device RBxposure Time Camera Binning and Sub Region Command Bar Cfg Acq Digital Camera Acquisition Parameters
25. MetaFluor Version 7 1 MetaFluor 5 MetaFluor Command bar New 1 Cfg Acd 2 Cfg Exp 3 Cfg Graphs Events Focus Set Disp Region Ratio References Moleoular Dewces apan KK Imggng Team 1 15 Cfg Acd Cfg Exp Cfg Graphs Events Focus Set Time Lapse Set Disp Region Ratio Reference
26. Ratio Ratio Display Min Display Max Ratio HMD Display Pseudocolor Monochrome IMD Display Ratio Rat in 1 es Image Display Contrals Win do IFati 1 lose Image Wirndom Properties Display ID Display IMD Intensity IMazamum of Waweleneths ID Overlay wavelength 1 Display in DD Display hiax IMD Display Ratio 1 75 Ratio 1 C75 Pseudocolor IMonochrome Moecular Deics apan KK Imggng Team 101 Region Ratio Region ROD 10 Command Bar Region SEleErt Image fnr efin img REgInrig Cfg Acd elect the irmaEa nh which to detrne the FEI 5 lmaEs Nia Fress the cquire ImaeEes button iT wou want to acquire arinthiar set of imaEes Acquire Images he tore yoau detne the FeEI TI Cfg Exp Cancel Select Image for Defining Regions Image
27. egion Threshold Create Region Region Min Size Max Size Regions Defined Region Active Region Region Average Intensity Region Region Done Molecular Deics apan KK Imggng Team 11 Command bar New Cfg Acd Cfg Exp Cfg Graphs Events Focus Set Time Lapse Set Disp Region Ratio References Command Bar References
28. s Command Bar New ms nmmand Har EieEIh E WW experiment pen st Fed az mieFirTiaht Lnad sprotocoal fle Play kawe From Disk rchiwe an esperiment to a ZF te rchie Extract Exit Command Bar More Less E tract IF fle to indivmdual files Et hetaFluor MetaFluor __ Neww 13 Pratocal ms nmmand Har Cotrol Focus Focus camers rne Acquire one set ofimaEes cg WW acquire igt Waweleneth 1 hc We acquire Inst Waweleneth cg WWH AEquira jiust Waweleneth 3 cd Wd cquire iust Waweleneth 4 C i EdUIFE st Waweleneth 5 Shutter Toeele selected shutter Reeginhns TEdit measurement reEions Eyents TEwent marks 1
29. seconds Load TIF Fth F ImraEa li mm h ane IF Path MM PP Imifl F raveleneth Letails Ewent hdarks quire TaEE FI Lndate ITiagg TE Cisplay Win dom TE awe MaEe hia TrigEer out Eposure Time EU milliseconds samera Gain 1 um De wce ue langth Intensitr se hutter WaveleneEth Waweleneth 1 Export MetaMorph Command Bar Fratacgl 1 Molecular Dewces apan KK Imggng Team 141 Load MetaFluor Command Bar Open Hile Open Experiment Load
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