Contents
Overview about
bh TCSPC Instrumentation......................................................................................................... 1
General Features of the bh TCSPC Devices.................................................................................................... 1
TSPC Module Types.......................................................................................................................................... 2
Measurement Modes of the bh TCSPC Modules.......................................................................................... 6
Simple-Tau TCSPC Systems.............................................................................................................................. 14
bh DSC‑120 Confocal Scanning FLIM
Systems............................................................................................. 14
bh FLIM Systems for Zeiss LSM 510 and
710 Laser Scanning Microscopes........................................... 15
bh FLIM Systems for Leica SP2 and SP5 Laser
Scanning Microscopes.................................................... 17
bh FLIM Systems for Olympus FV 300 and
FV 1000..................................................................................... 18
bh FLIM Systems for Nikon C1 scanners....................................................................................................... 19
DPC‑230 16 Channel Correlator / TCSPC
module........................................................................................... 19
PMS and MSA Multichannel Scalers.............................................................................................................. 19
Picosecond Diode Lasers.................................................................................................................................. 20
Detectors.............................................................................................................................................................. 21
Detector Electronics........................................................................................................................................... 24
Connecting Electronics...................................................................................................................................... 25
Instrument Software........................................................................................................................................... 26
Data Analysis Software..................................................................................................................................... 26
Classic Time-Correlated Single Photon
Counting................................................................................................ 29
Single-Photon Detection.................................................................................................................................... 29
Time-Correlated Single Photon Counting....................................................................................................... 29
The Classic TCSPC Architecture...................................................................................................................... 33
Multi-Dimensional TCSPC....................................................................................................................................... 37
Multi-Detector and Multi-Wavelength TCSPC.............................................................................................. 38
Multiplexed TCSPC............................................................................................................................................. 41
Sequential Recording......................................................................................................................................... 42
TCSPC Scanning FLIM Techniques................................................................................................................ 44
Time-Tag Recording........................................................................................................................................... 45
Imaging in the Time-Tag Mode........................................................................................................................ 47
Multi-Module TCSPC Systems......................................................................................................................... 48
Architecture of the bh TCSPC Modules................................................................................................................ 51
General Principle.................................................................................................................................................. 51
CFD and SYNC circuits................................................................................................................................ 51
TAC................................................................................................................................................................ 52
ADC................................................................................................................................................................ 52
Channel Register........................................................................................................................................... 53
Sequencer...................................................................................................................................................... 53
Memory Control............................................................................................................................................ 53
Recording of single curves................................................................................................................... 54
Multi-Detector Operation...................................................................................................................... 54
Multiplexed TCSPC................................................................................................................................ 55
Sequential Recording............................................................................................................................. 55
Continuous Flow Mode........................................................................................................................ 56
Scan Modes............................................................................................................................................ 56
CNT and ADD Control.......................................................................................................................... 57
Time-Tag (FIFO) Mode......................................................................................................................... 57
Detailed Description of Building Blocks......................................................................................................... 58
CFD in the Detector (Start) Channel.......................................................................................................... 58
SPC-x00 Versions.................................................................................................................................... 58
SPC-x30 Versions.................................................................................................................................... 59
CFD in the Synchronisation (Stop) Channel............................................................................................ 60
SPC-x00 Versions.................................................................................................................................... 60
SPC-x30 Versions.................................................................................................................................... 61
Time-to-Amplitude Converter..................................................................................................................... 62
Analog-to-Digital Conversion.................................................................................................................... 64
ADC Error Correction............................................................................................................................ 64
Variable ADC Resolution...................................................................................................................... 65
ADC Zoom.............................................................................................................................................. 66
Detectors for TCSPC................................................................................................................................................. 67
Detector Principles.............................................................................................................................................. 67
Conventional PMTs..................................................................................................................................... 67
Channel and Microchannel PMTs............................................................................................................. 67
Hybrid PMTs................................................................................................................................................. 68
Single-Photon Avalanche Photodiodes.................................................................................................... 69
Detector Parameters............................................................................................................................................ 70
Gain................................................................................................................................................................. 70
Single Electron Response............................................................................................................................ 71
Pulse Height Distribution............................................................................................................................ 72
Signal Transit Time....................................................................................................................................... 73
Transit Time Spread and Timing Jitter....................................................................................................... 73
Cathode Efficiency....................................................................................................................................... 74
Dark Count Rate............................................................................................................................................ 76
Afterpulsing.................................................................................................................................................. 77
Pre-Pulses...................................................................................................................................................... 78
Description of Selected Detectors.................................................................................................................... 80
MCP PMTs.................................................................................................................................................... 80
Hamamatsu R5600 and R7400 Miniature PMTs....................................................................................... 81
H5783 and H5773 Photosensor Modules.................................................................................................. 82
PMH-100 and PMC-100............................................................................................................................... 83
Hamamatsu H7422 and H8632..................................................................................................................... 84
Hamamatsu H7421........................................................................................................................................ 85
Hamamatsu R10467 Hybrid PMT............................................................................................................... 85
PML-16C Multichannel Detector Head..................................................................................................... 87
Conventional PMTs..................................................................................................................................... 88
Single-Photon Avalanche Photodiodes.................................................................................................... 89
Preamplifiers and Detector Control......................................................................................................................... 93
Preamplifiers......................................................................................................................................................... 93
The DCC-100 Detector Controller..................................................................................................................... 94
Electromagnetic Shielding................................................................................................................................. 96
Safety Recommendations for Using Detectors.............................................................................................. 100
Installation of the bh TCSPC Modules.................................................................................................................. 103
General Requirements......................................................................................................................................... 103
Software Installation........................................................................................................................................... 103
First Installation............................................................................................................................................ 104
Installing the DLL and Lab View Libraries................................................................................................ 105
Software Update........................................................................................................................................... 105
Update from the Web................................................................................................................................... 106
Installing New Software Components....................................................................................................... 106
Software Repair............................................................................................................................................. 107
Removing the TCSPC Package................................................................................................................... 107
Hardware Installation - Single SPC Modules.................................................................................................. 107
Driver Installation......................................................................................................................................... 107
Hardware Installation - Several SPC Modules................................................................................................ 108
Software Start...................................................................................................................................................... 108
Module Test Program......................................................................................................................................... 109
Installation Problems.......................................................................................................................................... 110
Starting the SPC Software without an SPC
Module...................................................................................... 110
Operating the SPC Module...................................................................................................................................... 113
Detector and Reference Inputs......................................................................................................................... 113
Generating the Reference Signal....................................................................................................................... 113
Delayed Stop Operation..................................................................................................................................... 115
Synchronisation to Random Signals................................................................................................................ 116
System Connection...............................................................................................................