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PHOTON COUNTING Using Photomultiplier Tubes INTRODUCTION TECHNICAL INFORMATION MAY. 1998 PHOTON COUNTING Using Photomultiplier Tubes INTRODUCTION Recently, non-destructive and non-invasive measurement using light is becoming more and more popular in diverse fields including biological, chemical, medical, material analysis, industrial instruments and home appliances. Technologies for detecting low level light are receiving par- ticular attention since they are effective in allowing high precision and high sensitivity measurements without changing the properties of the objects. Biological and biochemical examinations, for example, use low-light-level measurement for cell qualitative and quanlitative by detecting fluorescence emitted from cells labeled with a fluorescent dye. In clinical testing and medi- cal diagnosis, techniques such as in-vitro assay and im- munoassay have become essential for blood analysis, blood cell counting, hormone inspection and diagnosis of cancer and various infectious diseases. These techniques also involve low-light-level measurement such as colorim- etry, absorption spectroscopy, fluorescence photometry, and detection of light scattering or Iuminescence measure- ment. In RIA (radioimmunoassay) which has been used in immunological examinations using radioisotopes, radia- tion emitted from a sample is converted into low level light which must be measured with high sensitivity. In addition, fluorescence and Iuminescence measurements are used for rapid hygienic testing and monitoring processes in in- spections for bacteria contamination in water or in food processing. Photomultiplier tubes, photodiodes and CCD image sen- sors are widely used as “eyes” for detecting low level light. These detectors convert light into analog electrical sig- nals (current or voltage) in most applications. However, when the light level becomes extremely low so that the incident photons are detected as separate pulses, the single photon counting method using a photomultiplier tube is very effective if the average time intervals between sig- nal pulses are sufficiently wider than the time resolution of the photomultiplier tube. This photon counting method is superior to analog signal measurement in terms of stabil- ity, detection efficiency and signal-to-noise ratio. This technical manual explains how to use photomultiplier tubes in photon counting to perform low-light-level mea- surement with high sensitivity and high accuracy. This manual also describes the principle of photon counting, its key points and operating circuit configuration, as well as characteristics of photomultiplier tubes and their selec- tion guide. TABLE OF CONTENTS 1. Photon Counting ? ............................................................. 2 1-1 Analog Mode and Digital Mode (Photon Counting Mode) 1-2 The Principle of Photon Counting 2. Operation and Characteristics of Photon Counting ........ 5 2-1 Photon Counter and Multichannel Pulse Height Analyzer (MCA) 2-2 Basic Characteristics in Photon Counting (1) Pulse Height Distribution (PHD) and Plateau Characteristics (2) Output Instability vs. Variations in Photomultiplier Tube Gain (current amplification) (3) Linearity of Count Rate 3. Characteristics of Photomultiplier Tubes ......................... 9 3-1 Spectral Response (Quantum Efficiency:QE) 3-2 Collection Efficiency (CE) 3-3 Supply Voltage and Gain 3-4 Noise 3-5 Magnetic Shield 3-6 Stability and Dark Storage 3-7 Uniformity 3-8 Signal-to-Noise (S/N) Ratio 4. Measurement Systems ....................................................... 16 4-1 Synchronous Photon Counting Using Chopper 4-2 Time-Resolved Photon Counting by Repetitive Sampling 4-3 Time-Resolved Photon Counting by Multiple Gates 4-4 Time-Correlated Photon Counting (TCPC) 5. Selection Guide ................................................................... 18 5-1 Selecting the Photomultiplier Tube 5-2 Photomultiplier Tubes for Photon Counting 5-3 Related Products 1. Photon Counting ? Figure 1 : Output Pulses from Photomultiplier Tube at Different Light Levels TPHOC0027EA HIGHER LIGHT LEVEL (Multi Photoelectron State) 1-1 Analog Mode and Digital Mode ARRIVAL OF PHOTONS (Photon Counting Mode) PHOTOELECTRON EMISSION A photomultiplier tube (PMT) consists of a photocathode, SIGNAL OUTPUT (PULSE) an electron multiplier (composed of several dynodes) and an anode. (See Figure 2 for schematic construction.) When SIGNAL OUTPUT (DC) light enters the photocathode of a photomultiplier tube, TIME photoelectrons are emitted from the photocathode. These LOWER LIGHT LEVEL (Single Photoelectron State) photoelectrons are multiplied by secondary electron emis- ARRIVAL OF PHOTONS sion through the dynodes and then collected by the an- ode as an output pulse. In usual applications, these out- PHOTOELECTRON EMISSION put pulses are not handled as individual pulses but dealt with as an analog current created by a multitude of pulses SIGNAL OUTPUT (so-called analog mode). In this case, a number of pho- TIME tons are incident on the photomultiplier tube per unit time as in of Figure 1 and the resulting photoelectrons are emitted from the photocathode as in . The photoelec- 1-2 The Principle of Photon trons multiplied by the dynodes are then derived from the anode as output pulses as in . At this point, when the Counting pulse-to-pulse interval is narrower than each pulse width or the signal processing circuit is not fast enough, the ac- One important factor in photon counting is the quantum tual output pulses overlap each other and become a di- efficiency (QE). It is the production probability of photo- rect current with shot noise fluctuations as shown in. electrons being emitted when one photon strikes the pho- In contrast, when the light intensity becomes so low that tocathode. In a single photoelectron state, the number of the incident photons are separated as shown in, the out- emitted photoelectrons (primary electrons) per photon is put pulses obtained from the anode are also discrete. This only 1 or 0. Therefore QE refers to the ratio of the average condition is called a single photoelectron state. The num- number of emitted electrons from the photocathode per ber of output pulses is in direct proportion to the amount unit time to the average number of photons incident on of incident light and this pulse counting method has ad- the photocathode. vantages in signal-to-noise (S/N) ratio and stability over the analog mode in which an average of all the pulses is Figure 2 : Photomultiplier Tube Operation made. in Single Photoelectron State This pulse counting technique is known as the photon PHOTOCATHODE 1ST DYNODE counting method. Since the detected pulses undergo bi- ANODE nary processing for digital counting, the photon counting P method is also referred to as the digital mode. PULSE HEIGHT SINGLE PHOTON Dy1 Dy2 Dyn-1 Dyn ELECTRON GROUP TPMOC0048EB 2 Photoelectrons emitted from the photocathode are accel- Figure 3 : Photomultiplier Tube Output and PHD erated and focused onto the first dynode (Dy1) to produce secondary electrons. However, some of these electrons do not strike the Dy1 or deviate from their normal trajecto- LOW COUNT FREQUENCY ries, so they are not multiplied properly. This efficiency of LOW COUNT FREQUENCY HIGH COUNT FREQUENCY collecting photoelectrons is referred to as the collection efficiency (CE). In addition, the ratio of the count value (the number of output pulses) to the number of incident photons is called the detection efficiency or counting effi- PULSE HEIGHT ciency, and is expressed by the following equation : COUNTS AT EACH Detection efficiency(%) =(Nd/Np) × 100(%) = η × α × 100(%) where Nd is the count value, Np is the number of incident photons, η is the photocathode QE and α is the CE. Al- though discussed later, detection efficiency also depends TIME on the threshold level that brings the output pulses into a binary signal. Since the number of secondary electrons emitted from the Dy1 varies from several to about 20 in response to one primary electron from the photocathode, PULSE HEIGHTS (CHARGES) TPMOC0049EB they can be treated by Poisson distribution, and the aver- age number of electrons becomes the secondary electoron δ emission ratio . This holds true for multiplication processes Photomultiplier Tube Output in Single in the subsequent dynodes. Accordingly, for a photomulti- Photoelectron State plier tube having n stages of dynodes, a single photoelec- The output signal from a photomultiplier tube in the pho- tron from the photocathode is multiplied by δ n to create a ton counting mode can be calculated as follows: group of electrons and is derived from the anode as an In the photon counting mode, a single photoelectron e- output pulse. In this process, the height of each output (electron charge 1.6 × 10-19 coulombs) is emitted from pulse obtained at the anode depends on fluctuations in the photocathode. If the photomultiplier tube gain µ is 5 the secondary electron multiplication ratio stated above, × 106, then the anode output charge is given by so that it differs from pulse to pulse. (Figure 3) e × µ = 1.6 × 10-19 × 5 × 106 (coulombs:C) Other reasons why the output pulse height becomes un- = 8 × 10-13 (C) equal are that gain varies with the position on each dyn- Here, if the pulse width t (FWHM) of the anode output ode and some deviated electrons do not contribute to the signal is 10ns, then the output pulse peak current Ip is normal multiplication process. Figure 3 shows a histogram Ip = e × µ × 1/t (A) of the anode pulse heights. This graph is known as the = (8 × 10-13)/(10 × 10-9) pulse height distribution (PHD).
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