Cornelius DPC 230 Specifikace

High PerformancePhoton CountingDPC-23016 ChannelPhoton CorrelatorUser HandbookBecker & Hickl GmbH

4 Principle of Data Acquisition Three of the CFD channels are used for the detector signals. The CFD output pulses of these channels are fed dire

Operation Modes Absolute-Timing Modes In the ‘Absolute Time’ mode every photon is characterised by its time from the start of the measurement and it

6 Operation Modes tive brightness and the concentration ratio of the molecules to the measured PCH. The tech-nique is also called ‘fluorescence i

Absolute-Timing Modes 7 The autocorrelation (shown left) has a sharp peak at τ = 0. (The function correlates perfectly with itself). The slow fluct

8 Operation Modes The procedure illustrated in Fig. 10 yields G(τ) in equidistant τ channels. The width of the τ channels is equal to the time-ch

Relative Timing Modes 9 of the excitation source, and the distribution of the events within the excitation period is built up. Thus, the multichann

10 Operation Modes result is the waveform of the light signal. Because three input channels are available in the TCSPC mode three signals from th

Relative Timing Modes 11 Other applications of the absolute times in TCSPC data are multi-parameter single-molecule spectroscopy [19, 25, 31], burs

12 Operation Modes ton count rate. This makes the recording process more or less random. The recording is con-tinued over as many frames as neces

Installation Computer In principle, the DPC-230 module can be installed in a PCI slot of any Pentium PC. However, the SPCM software runs the data tr

(c) Becker & Hickl GmbH

14 Installation Fig. 16: Installation panels The installation works the same way as the installation from the CD. Check the boxes of th

Software Start 15 Fig. 17: Downloading the drivers from the bh web site Open www.becker-hickl.com and click on ‘Software’. On the ‘Software’ pa

16 Installation Starting the SPCM Software without a DPC-230 Module You can use the Multi SPC Software without a DPC module. In its start window

Operating the DPC-230 LVTTL Inputs The LVTTL inputs of the DPC-230 are designed to receive single-photon pulses from SPAD (single-photon avalanche p

18 Operating the DPC-230 Manufacturer Type Pulse Pulse Remark Amplitude Width id Quantique id 100-xx +2 V 20 ns Versions of different ar

CFD Inputs 19 Connecting PMTs to the CFD Inputs PMTs use secondary-electron emission to multiply a single photoelectron by a factor of 106 to 108.

20 Operating the DPC-230 Manufacturer Type Preamplifier HV / Gain CFD Thresh. Remark bh PMC-100-201) internal 100 %2) -40 mV cooled PMT

Typical Applications Fluorescence Decay Measurements A typical experiment setup for fluorescence decay measurement is shown in Fig. 26. The sam-ple

22 Typical Applications The right part of the panel shows the discriminator parameters of the CFD inputs and the TDC parameters. The optimal disc

Luminescence Decay Measurement in the Microsecond Range 23 Fig. 29: Trace parameters and display parameters recommended for fluorescence d

Becker & Hickl GmbH April 2008 High Performance Photon Counting DPC-230 16 Channel Photon Correlator Photon correlation down to the ps ran

24 Typical Applications The luminescence light is detected the same way as for fluorescence decay measurement. The reference pulses for the DPC-2

Fluorescence Correlation 25 A typical optical setup is shown in Fig. 34. A CW laser beam is focused into the sample through a microscope objective

26 Typical Applications stopped after a defined acquisition time. Activate the ‘Stop T’ button if you want to stop after a specified time. ‘Max B

Fluorescence Correlation 27 The curves to be displayed in the display windows for the accumulated counts, FCS curves, and MCS traces are defined un

28 Typical Applications Fig. 40: Cross-Correlation between two detector channels Picosecond Fluorescence Correlation Fluorescence correlation

Anti-Bunching 29 Anti-Bunching Anti-bunching information is contained in picosecond fluorescence correlation data, see Fig. 41. However, anti-bunch

30 Typical Applications Fig. 43: Input configuration for a classic anti-bunching start-stop experiment To see the desired start-stop histograms,

Fluorescence Lifetime Imaging 31 Fig. 46: Start-stop histogram and ps correlation obtained in the same measurement Fluorescence Lifetime Imaging

32 Typical Applications The scan clock pulses (pixel clock, line clock, frame clock) of the microscope are connected into three LVTTL channel of

Fluorescence Lifetime Imaging 33 the FLIM image. Moreover, many microscopes send a frame clock pulse some pixels before the start of the useful par

II Becker & Hickl GmbH Nahmitzer Damm 30 12277 Berlin Germany Tel. +49 / 30 / 787 56 32 FAX +49 / 30 / 787 57 34 http://www.becker-hickl.

34 Typical Applications Fig. 52: Fluorescence lifetime image recorded with DPC-230 and bh DCS-120 confocal scanning FLIM system [4]. Data analys

Luminescence Lifetime Imaging in the Microsecond Range 35 beginning of each pixel. The pixel time is made long enough to observe the full luminesce

36 Typical Applications Fig. 56: Input configuration panel. Left: PMT on CFD 3, reference on CFD 4. Right: SPADs on LVTTL 5 through 2

SPCM Software The DPC-230 comes with the ‘Multi SPC Software’, or ‘SPCM’ operating software. The SPCM software is not only used for the DPC-230 but

38 SPCM Software A typical main panel of a correlation measurement in the ‘Absolute Time’ mode is shown in the upper row, left. It shows the phot

Configuring the SPCM Main Panel 39 The Display Parameters (shown left) allow you to define how your results will be displayed. The upper part refer

40 SPCM Software Fig. 62: Select panel for display size, cursor display, and display and trace parameters It can happen that a

Configuring the SPCM Main Panel 41 instrument configurations several data sets may have been recorded. In this case click on the display window tha

42 SPCM Software lost. Strictly, such data cannot be correlated any more. However, as long as only a few over-flows occurred correlation within o

System Parameters of the DPC-230 43 System Parameters of the DPC-230 The system parameters panel of the DPC-230 is shown in Fig. 70. The panel cont

Contents III Contents Introduction...

44 SPCM Software next reference pulse (see ‘Relative Timing’, page 3). The data of the detector channels can be interpreted either as single wave

System Parameters of the DPC-230 45 full. Although this may slow down the data transfer from the SPC module the loss is by far smaller than for mem

46 SPCM Software - Calculation of photon counting histograms for the individual detectors (‘FIDA’). The sampling time interval is specified on t

System Parameters of the DPC-230 47 Both TDC chips can be switched on and off by the ‘active’ button. If one of the TDCs is not used it should be s

48 SPCM Software Fig. 79: SYNC frequency divider setting. The parameters determines the number of signal periods recorded in the TCSPC mode TDC

Saving Setup and Measurement Data 49 Saving Setup and Measurement Data The ‘Save’ panel is shown in Fig. 81. It contains fields to select different

50 SPCM Software culated data or data loaded from another file. Except for special cases (see[2]) we recommend to use the ‘All used data sets’ op

Predefined Setups 51 Block Info Activating a data block in the ‘Block Number in File’ field enables a ‘Block Info Button’. Clicking on this button

52 SPCM Software Fig. 85: Editing the list of predefined setups To create your own predefined setups, first save a setup file of the system conf

Importing FIFO Files 53 Importing FIFO Files Measurements in the absolute time modes deliver an .spc file that contains time-tag data, i.e. the mic

IV Loading Setup and Measurement Data ...50 Predef

54 SPCM Software The ‘Convert FIFO’ routine allows you to convert .spc files into different destination file types. The destination file type is

Format of Time-Tag Data Files 55 Format of Time-Tag Data Files In most of the operation modes the DPC-230 allows the user to record time-tag data o

56 SPCM Software loss of photons or any loss in photon information. Pre-processed data are identified by ‘RAW = 0’. The description given below r

Format of Time-Tag Data Files 57 High-Time Record A record with bit 30 = 1 and bit 31 = 0 indicates that the higher part of the time has changed, a

Specification LVTTL Inputs No. of channels 16 Input Voltage LVTTL Threshold 1.4 V Min. Input Pulse Width 2 ns Min. Pulse Distance 5.5 ns Connec

References 1. W. Becker, Advanced time-correlated single-photon counting techniques. Springer, Berlin, Heidelberg, New York, 2005 2. W. Becker, T

62 References 28. R. Rigler, J. Widengreen, Utrasensitive detection of single molecules by fluorescence correlation spec-troscopy, Bioscience 3,

Index 2D FIDA 8 Absolute timing 2 Absolute timing mode 7 definition in the system parameters 45 Anti-bunching 31 combined with ps FCS 32 softw

Introduction The DPC-230 photon correlator card records absolute photon times in up to 16 parallel detec-tion channels. Depending on how the photons

64 Index Inputs CFD inputs 20 configuration of 49 for PMT modules with TTL output 19 for PMT pulses 21 for SPADs 19, 21 LVTTL inputs 19 mar

2 Principle of Data Acquisition ring is read, and used to determine the detection time. Times longer than the reference cycle time are determined

Principle of Data Acquisition 3 When a measurement is started the TDCs in all active channels simultaneously start running. Any event detected at o