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...............................................................................................................