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Transcription:

MetaFluor (Version 7.7) MetaFluor 1. MetaFluor MetaFluor Meta Imaging Series 7.x Meta Imaging Series Administrator CCD Meta Imaging Series Administrator CCD Molecular Devices Japan KK/ Imaging Team (1/14)

2. MetaFluor Meta Imaging Series 7.x MetaFluor 3. MetaFluor Illumination Configure Illumination Control (Configure Illumination) Ludl (Ludl Wheel 1)(Ludl Wheel 2) (Ludl Shutter 1)(Ludl Shutter 2)FRET-CFP, FRET-YFP, DAPI, FITC, Rhodamine, BF Name: Add / Replace Illumination Setting Add / Replace Defined Settings Backup Illumination Settings Molecular Devices Japan KK/ Imaging Team (2/14)

4. New File New Experiment More ( Cfg Acq) Configure Configure Acquisition Configure Acquisition (Illumination Device)(Exposure Time)(Camera Binning and Sub-Region) Digital Camera Acquisition Parameters. Wavelength 1. Illumination Devices: Configure Illumination Illumination. Exposure Time, Gain. Binning (Sub-Region) Select Region Use Entire Image. Wavelength Z Set Z Position for this Wavelength to : Z Z MetaFluor. Wavelength 2 Allow items to differ for each wavelength Digital Camera Acquisition Parameters Wavelength Wavelength Molecular Devices Japan KK/ Imaging Team (3/14)

( Control) Run Experiment Control Experiment Panel Experiment Control Panel (F4: Acquire)(Set Timelapse)(Focus) F4: Acquire F2: Pause F4: Acquire Zero Clock () Set Timelapse... Set Timelapse (Timelapse Interval) (Number of Acquisitions) Timelapse Interval Number of Acquisitions 101 101 Number of Acquisitions (Pause) Log Data Regions Configure Experiment (Cfg Exp)Ave, Sum, Area Measurements Dynamic Data Exchange (DDE) Microsoft Excel Excel OK Excel A Text File Log OK Molecular Devices Japan KK/ Imaging Team (4/14)

Save Image.INF inf 001 inf, inf Save Ratios Ratio Ratio TIF TIF Log Data, Save Ratios Molecular Devices Japan KK/ Imaging Team (5/14)

Focus Focus XY Start Focusing Stop Focusing Illumination: Wavelength Configure Acquisition Configure Acquisition Wavelength Illumination Wavelength (Camera Settings:, Exp:) (Acquisition Region:) Store for this Wave, Store as Acq Rgn Configure Acquisition, CCD Configure Acquisition Exposure Time Camera Binning and Sub-Region Start Focusing Wavelength: Stop Focusing Z Z 0 Goto 0 Store for this Wave Configure Acquisition Camera Settings: Close Focus MetaFluor Histogram of Image Molecular Devices Japan KK/ Imaging Team (6/14)

( Cfg Exp) Configure Configure Experiment Configure Experiment Acquire? / Acquire Interval: 1 2 2 1 Data Logging: Experiment Control Panel Log Data AveSumArea Regions Save Interval: Experiment Control Panel Save ImageRatio 1, 2 Save Ratio Show? / Update? / Update Interval: Update Ratio 1, Ratio 2, Calcium Ratio(W1 / W2)Ratio(W4 / W5)Calibrated Ratio 1/ ON/OFF (Wavelength)/ Wavelength 1(FRET-YFP), Wavelength 2(FRET-CFP) Wavelength 1(FRET-YFP), Wavelength 2(FRET-CFP), Ratio 1(W1/W2) MetaFluor Wavelength 1Wavelength 2Ratio 1 Show?, Update?, Update Interval, Save Interval, Ratio (Frames per Second) Ratio Molecular Devices Japan KK/ Imaging Team (7/14)

( Regions) Run Experiment Select Save Regions... (Region, ROI) Select Image for Defining Regions Image: Region Region Acquire Images 1 OK Region Tool Region Edit Regions Region (Save) (Load) Threshold Create Region Done Cfg Graphs Molecular Devices Japan KK/ Imaging Team (8/14)

Cfg Graphs Graphs Configure Graphs Configure Graphs Appears on Graph: Wavelength 1, 2, 3, 4, 5 Average Intensity None, Graph 1, 2, 3, 4, 5 Line Type: Solid, Wide, Dotted, Dashed Graph 1, 2, 3, 4, 5 Show Graph / Graph: X Y Graph Setting Molecular Devices Japan KK/ Imaging Team (9/14)

( Event) Run Experiment Event Marks MetaFluor Event Marks Mark New Event Now Inf Setting for New Event Event Text: Add New Event to List F5: Mark F5, F6 Molecular Devices Japan KK/ Imaging Team (10/14)

( Set Disp) Configure Image Display Control Wavelength 1 (Brightness) (Contrast) Display Monochrome, Pseudocolor Ratio Ratio (Display Min:)(Display Max:)Ratio (IMD Display, Pseudocolor, Monochrome)IMD Display Ratio IMD Display Pseudocolor Monochrome Molecular Devices Japan KK/ Imaging Team (11/14)

5. ( Open) File Open Experiment File Close ExperimentClose INF (O) Experiment Control Panel Event List Experiment Control Panel (Frame:)Image Time: F3: Forward, F3: Reverse, F2: Pause Retio Event List Molecular Devices Japan KK/ Imaging Team (12/14)

6. ( References) Run Experiment Reference Imges Reference Images Wavelength Background Shaging Image Load Region Parameter Region Region Region Subtract Backgrounds Correct Shading Region 1 Parameter Region Subtract Background Wavelength 1, 2 Region 7. Utilities Play Movie from Disk Playback a Movie Stored on Disk Select Images Select First Image Select Last Image Next >> 2 Play Movie from Disk Save As AVI Molecular Devices Japan KK/ Imaging Team (13/14)

8. MetaFluor MetaFluor ZIP File Archive Experiment Source File Directory Archive Location Source Files INF Archive Description Archive Molecular Devices Japan KK/ Imaging Team (14/14)