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**Optical Spectroscopy** bridges a gap by providing a background on optics while focusing on spectroscopic methodologies, tools and instrumentations. The book introduces the most widely used steady-state and time-resolved spectroscopic techniques, makes comparisions between them, and provides the methodology for estimating the most important characteristics of the techniques such as sensitivity and time resolution.

Recent developments in lasers, optics and electronics has had a significant impact on modern optical spectroscopic methods and instrumentations. Combining the newest lasers, advanced detectors and other high technology components researchers are able to assemble a spectroscopic instrument with characteristics that were hardly achievable a decade ago. This book will help readers to sourse spectroscopy tools to solve their problems by providing information on the most widely used methods while introducing readers to the principles of quantitative analysis of the application range for each methodology. In addition, background information is provided on optics, optical measurements and laser physics, which is of crucial importance for spectroscopic applications.

* provides an overview of the most popular absorption/emission spectroscopy techniques

* discusses application range, advantages and disadvantages are compared for different spectroscopy methods

* provides introductions to the relevant topics such as optics and laser physics

Publisher: Elsevier ScienceReleased: Jun 6, 2006ISBN: 9780080461724Format: book

This book is intended for graduate and post graduate students, and researchers planning to start an advanced experimental work in the fast growing field of optical spectroscopy. Optical spectroscopy methods have numerous applications in physics, chemistry, material, biological and medical sciences. Probably two most exciting achievements of the optical spectroscopy are single molecule detection and ultrafast time resolution. The former shifts research work to a molecular scale and serves as the key tool in areas known as nanochemistry and molecular devices. The latter extends the time scale to femtoseconds making possible direct studies of chemical reactions on the level of the chemical bond dynamics at the atomic scale. This research field is commonly called Femtochemistry after Nobel price winer Ahmed H. Zewail.

The progress in the optical spectroscopy was possible because of great developments in laser physics, optics, electronics and computers. Combining together newest lasers, advanced detectors and high technology optical components a researcher can assemble relatively easy a spectroscopy instrument with characteristics which were hardly achievable in the top level laser research labs a decade ago. Naturally, the first step in this direction is to be informed on the tools available and to be able to evaluate the benefits and limitations imposed by different techniques. On the other hand the researchers, who are potentially interested in such spectroscopy applications, are experts in the fields of their own professional interest, such as materials science or microbiology, and may have only basic background knowledge in optics and modern laser physics. The aim of the book is to cover the gap by providing background information in optics and by focusing on spectroscopy methodology, tools and instrumentations. The goal is to provide a background for quantitative estimations of the applicability range of optical spectroscopy methods and to help researchers in planning, designing and developing of new spectroscopy instruments, and, hopefully, new spectroscopy methods.

Logically the book can be divided on two parts. The first part, **Chapter 1–4, covers a few subjects important for technical implementations of all spectroscopy instruments. This includes optics, opto-electronics, laser physics and related topics. This part of the book has no intention to go deep into its subjects nor to provide a complete overview of these broad areas. In turn, it is supposed that readers are already familiar with the subject and the main goals of this part of the book is to remind readers of the key concepts, important theories, principle values and available tools which are used in spectroscopy applications. One example of such tool is a monochromator. Monochromators can be found in almost every spectroscopy device and they are crucial for such important characteristics of the devices as spectrum resolution and sensitivity. **

The second part of the book, **Chapter 5–13, is the main part, which divided onto Chapters according to the types of the optical spectroscopy methods described. Each Chapter starts with a general description of the design principles of a particular methods. This follows by an introduction of the approaches used to estimate the most important features of the instrument, such as spectrum and time resolution, and by discussions of what are applicability ranges for this particular spectroscopy technique. In the final part of each Chapter examples of the instruments and measurements are provided. The order of the Chapters roughly follows the order of inventing different spectroscopy methods and one can see that the important direction in the development of optical spectroscopy instruments was the continuous improvement of the time resolution. **

It is important to notice from the very beginning that the spectroscopy techniques do not exist for its own – they were developed for the purpose of investigation of certain objects, e. g. natural photosynthetic centers. The application of one or another method, or combination of methods is justified by the object to be studied and the problem to be solved. Therefore one of the main goals of the book is to compare different spectroscopy techniques and to highlight advantages and disadvantages of them in respect to the most common application tasks.

Two last topics discussed in the book are polarization (**Chapter 14) and analysis of the measurements (Chapter 15). They are important to all spectroscopy techniques and, therefore, were arranged as separated Chapters. The polarization of the light is its fundamental property. If it is ignored, it may lead to misinterpretation of the experimental results. On the other hand, a careful accounting for the light polarization and sample anisotropy may help to improve the quality of the measured data. Additionally, polarization and anisotropy can be used to gain an additional information on the samples under study. For example, the excitation energy transfer can be studied by measuring sample anisotropy. In the latter case the actually measured data are light intensity time profiles at different polarizations of the light. The anisotropy is calculated then out of these primary measurements. This example highlights the importance of the accurate analysis of the experimental data. Even more common data analysis problem for the optical spectroscopy is extraction of the lifetimes of the intermediate states in a photochemical reaction. This is the subject of Chapter 15. **

At last, but not at least, I would like to express my gratitude to many great scientist who were my teaches and my colleagues. After graduation from Moscow Institute of Physics and Technology I have joined General Physics Institute (GPI) as young researcher, where I was guided by Prof. A. M. Prokhorov and Dr. V. V. Savranskii in my work and doctor thesis preparation. From them I have learned a great deal about laser physics. In GPI I have assembled my first flash–photolysis instrument together with A. Dioumaev, V. Chukharev and A. Sharonov, young scientists at that time. Later I was invited by Prof. H. Lemmetyinen to join his team at the University of Helsinki. Prof. Lemmetyinen is a great experts in photochemistry, and that was the time to learn new optical spectroscopy methods, such as the time correlated single photon counting. A few year later the research team moved to Tampere University of Technology, where the instrumentation facilities of the group were extended by building up a new femtosecond spectroscopy system to be able to carry out optical spectroscopy studies in a wide time scale from steady state to femtoseconds. Educating students for advanced research work in the spectroscopy laboratory was the motivation to write this book.

Most of all, I am indebted to my family, especially my wife Natalia for her love and tolerance of irregular hours at home, and my daughter Evgenia for her help in text preparation.

**Nikolai V. Tkachenko, ***Tampere, Finland *

December 2005

Optical spectroscopy studies absorption and emission of light by matter. Originally the studies were related to the wavelength dependences of these processes, but as new methods and directions of research were developed the scope of optical spectroscopy has enlarged. One of the important directions of such development today is time resolved spectroscopy, where optical methods provide superior time resolution not achievable by any other methods available. There are also specialized areas of optical spectroscopy such as single molecule spectroscopy and non-linear optical spectroscopy, which have also been under active development during the past decade.

Naturally, the reason for the great attention paid to the optical spectroscopy techniques in recent fundamental research is new exciting knowledge gained. To mention few there are chemical reaction dynamics at single bond level, called femtochemistry after Ahmed H. Zewail, and single molecule spectroscopy. Furthermore, non-destructive spectroscopy methods have found numerous applications in monitoring different processes in industry and environmental technologies.

The aim of this book is to give an overview of the modern optical spectroscopy methods, and to introduce the principles used to evaluate quantitatively advantages and application ranges of the methods. The author hopes that this information will help researchers and application engineers to plan their optical spectroscopy work and to get the most of each method used.

In this introductory chapter we will briefly look at fundamental concepts of the light absorption and emission. The main goal, however, is to mention the most widely used concepts and terms in optical spectroscopy and its applications. It is assumed that the readers are quite familiar with the subject and a short reminding of the topic can be appropriate for the following consideration.

Let us consider a beam propagating in an isotropic dielectric medium. One can choose a coordinate system so that the beam is propagating along X-axis, as presented in **Fig. 1.1. At a point x the light intensity is I(x). In a layer of thickness Δx the light interacts with the medium and at the point x + Δx the light intensity is changed by value ΔI, which means I(x + Δx) = I(x) + ΔI(x, Δx). At Δx is proportional to the light intensity: **

**Figure 1.1 **Light absorption in a medium.

**(1.1) **

or

**(1.2) **

where α is the proportionality coefficient determining efficiency of the light absorption. The minus sign on the right side of the equation is due to the fact that the light is absorbed by the medium, i. e. the function *I*(*x*) is decreasing with increasing *x*, and, thus, it has a negative slope. To verify the statement (**1.1), let us consider a transparent medium which incorporates absorbing centers, for example dye molecules in a solution. The probability, p, to absorb a photon by a thin layer of the medium is proportional to the surface density of the absorbing centers, s, and absorption efficiency of the centers, σ, so that p = sσ. If the volume density (concentration) of the absorbing centers is n, then the surface density of the absorbing centers in the layer of thickness Δx is s = nΔx. For a very thin layer the absorption probability is very small positive value, and for the incident light flux I, the decrease in intensity is ΔI = −Ip, or –ΔI = Isσ = InσΔx. , where α = nσ. **

**Equation (1.2) can be rearranged **

**(1.3) **

and solved

**(1.4) **

Where the constant *C *is determined by the initial conditions, which is the incident light intensity at *x *= 0, i. e. *I*(0) = *I*0, in this particular case. **Equation (1.4) is usually converted to the form known as Lambert law¹ **

**(1.5) **

The coefficient α is called absorption coefficient and it has measure of inverse length (e. g, cm−1). If the coefficient α > 0, then the light intensity decreases along the light propagation direction. There is no light absorption if the coefficient α = 0, i. e. *I*(*x*) = *I*0 = *constant. *The light intensity also increases exponentially if α < 0. The latter case is called light amplification and will be discussed in **Section 1.3. **

When the light propagate across finite length absorbing medium, one may be interested in portion of the light which will be absorbed or will pass the medium. Let us assume that the thickness of the medium is *l *(**Fig. 1.1) and its absorption coefficient is α. The light intensity before the medium is Iin = I0 = I(0) and, according to eq. (1.5), the light intensity after the medium is **

**(1.6) **

The transmittance of the sample (the relative amount of the light passing through the sample) is

**(1.7) **

In other words, **eq. (1.6) can be rewritten as **

**(1.8) **

Consequently, the absorptance of the sample (the relative amount of the light absorbed by the sample) is

**(1.9) **

These values, absorptance and transmittance, are usually used when one needs to calculate intensity of the light propagating in some optical system, i. e. when the light intensity distributions inside the optical components are out of interest.

The absorptance and transmittance are not very convenient characteristics when someone is interested in the optical properties of the absorbing centers, since the absorptance depends on the thickness of the sample and the density of centers. Because of some practical and historical reasons different values are used to characterize absorption properties of media. The absorption coefficient, α, as given by **eq. (1.6), is a common specification for the media such as glasses or fibers.² The absorption coefficient does not depend on optical path and characterizes the medium itself. **

Absorption coefficient of a fiber can be 0.001 m−1, which means that 1 km of the fiber will absorb 1 – exp(−10−3 · 10³) = 1 – exp(−1) ≈ 0.63 = 63% of the incoming light power. A gray filter HC-3 has absorption coefficient ≈1 mm−1 in the visible part of the spectrum, thus transmittance of the filter of 2 mm thickness is exp(−2) ≈ 0.14 = 14%.

When the light absorption is considered at a molecular level, the absorption cross-section, σ, is a more practical value to be used since it characterizes a single molecule and does not depend on the density (concentration) of the molecules. Then, **eq. (1.6) is presented in the form **

**(1.10) **

where *n *is the density of the absorbing centers (e. g. dye molecules).**³ **

In many practical cases power of 10 is used instead of power of *e. *Then, **eq. (1.6) is rewritten to the form **

**(1.11) **

where *A *is the absorbance or optical density. Naturally, the absorbance can be calculated from the light intensities at the sample input and output

**(1.12) **

Comparing **eqs. (1.7) and (1.12) one obtains a relation between the transmittance and absorbance **

**(1.13) **

,**⁴ which is the proportionality coefficient in the relation **

**(1.14) **

**Note**: By historical tradition the measure of molar concentration is moles per liter, i. e. M = mol×dm−3, whereas optical path, *l*, is counted in cm, thus the measure of the molar absorption coefficient is M−1cm−1 = mol−1dm³cm−1, i. e. one value uses lengths measured in different units, dm and cm!

The molar absorption coefficient and the cross-sections are two characteristics specifying the same property of chromophores, their ability to absorb the light. Comparing **eqs. (1.10), (1.9) and (1.14), and recalling that c = n/NA, where NA is the Avogadro constant, one obtains **

**(1.15) **

which gives

**(1.16) **

in M−1cm−1.

Absorption cross-section of chlorophyll *a *M−1cm−1. A chlorophyll solution at concentration *c *= 10−5 M = 10 μM placed in *l *= 1 cm cuvette will have absorbance *A **cl *= 1, and will absorb 1 − 10−*A *= 1 − 0.1 = 0.9 = 90% of the incident light.

In summary, there are few parameters, which can be used to characterize the light absorption by the matter. The usage of the particular parameter depends on the problem on hands. The parameters can be

• absorptance, *a*, or transmittance, *T*, which are dimensionless values and usually expressed in % (**eqs. (1.7) and (1.9)); **

• absorbance (optical density), *A*, which is dimensionless parameter, **eq. (1.11); **

• absorption coefficient, α, which has dimensionality of inverse length, e. g. cm−1, **eq. (1.6); **

• absorbtion cross-section, σ, is used to specify absorption properties of a single molecule, for example, and is measured by the area, e. g. cm², **eq. (1.10); **

, which is usually used to specify absorption properties of chemical compounds and has dimensionality of M−1cm−1, **eq. (1.14). **

(λ), which is the measurement of the absorption spectrum.

Let us now consider a practically important case of a complex sample consisting of a few layers with different absorbing properties. This can be a solution of some compound in a cuvette, for example. Typically the absorption spectrum of the compound is of interest, but it is possible that in the wavelength range of interest the solvent and the cuvette have absorptions of their own. To simplify the case, the sample can be presented as a sequence of absorbing layers, as shown in **Fig. 1.2. In front of the sample, behind layer 1, the light intensity is I0. The transmittance of the first layer is T1, thus the light intensity after the layer is I1 = I0T1. The light intensity entering layer 2 is I1. The transmittance of the layer 2 is T2, which gives the light intensity at the interface between layer 2 and layer 3, I2 = I1T2 = I0T1T2. Continuing this procedure, the light intensity after the layer 3 is I3 = I2T3 = I0T1T2T3. This gives the total transmittance of the sample T = T1T2T3, i. e. the product of the transmittances of the individual layers. **

**Figure 1.2 **Absorption of multi layer sample.

In a more general case, the transmittance of a complex sample is the product of the transmittances of the individual components forming the sample

**(1.17) **

where *N *is the number of absorbing components, layers in the above example. This is equally applied to a mixture of dye molecules in solution if there is no intermolecular interaction. If the transmittance of one dye in a cuvette at some concentration is *T*1 and the transmittance of another dye in the same cuvette is *T*2, then the mixture of the dyes will have transmittance *T *= *T*1*T*2, under condition that the concentrations of the dyes are the same as in the individual measurements.**⁵ **

To calculate absorbance of a complex sample one can apply **eq. (1.13) to eq. (1.17) to obtain **

**(1.18) **

This is a simple and practically important result – the absorbance of a complex sample is the sum of absorbances of its components. Returning back to the example discussed in the beginning of this Section we can conclude that to obtain the absorption spectrum of the compound of our interest we need to measure the absorbance of the sample solution, then the absorbance of the same cuvette filled with the same solvent but without the compound, and after subtracting the latter from the former we will obtain the spectrum of the compound. More discussion in this subject follows in the **Chapter 5. **

At a molecular level, the light absorption results in a change of the molecule state. This is usually discussed in terms of energy levels and transitions between them since the molecules are quantum objects. Depending on which part of molecular subsystem is involved, the energy levels are divided onto electronic, vibrational and rotational.

The electronic levels are associated with the energy of the electron subsystem. The transition form one electronic level to another can be considered as the transition of one of the electrons form one orbital to another. The vibrational levels are related with the vibrational motions of the molecules. The transitions between them have almost no effect on the electron subsystem. The rotational levels arise from rotations of molecules as whole.

for harmonic oscillators). Typical ranges of the transition are summarized in **Table 1.1 and presented graphically in Fig. 1.3. **

**Table 1.1 **

**Energies and spectral ranges of different types of transitions. **

**Figure 1.3 **Relations between different scales used in spectroscopy: wavelength, λ, wave number, *k*, and frequency, *v. *Ultraviolet (UV), visible and near infra-red (NIR) parts of the spectrum are indicated in an enlarged part of the wavelength scale.

The wavelength range of the visible light is 0.4–0.7 μm. Accounting for the near infrared region (0.7–1.5 μm) and ultraviolet (UV) (0.2–0.4 μm), the optical wavelength range can be considered to be from 0.2 to 1.5 μm. Therefore, strictly speaking, only electronic transitions fall into the optical range. The vibrational and rotational transitions corresponds to the infrared and far infrared wavelength regions.

However, all the levels contribute to the absorption spectrum of the molecules in the visible and UV ranges. The electronic levels determine positions of the absorption bands and the vibrational and rotational levels contribute to the shapes of the bands. In addition, the shapes of the bands are affected by numerous line broadening

mechanisms.**⁶ **

Since the energy of a photon determines frequency (and wavelength) of the electromagnetic wave (by famous Planck formula *E *= *hv *in vacuum), the absorption spectrum of any system represents its energetic spectrum or density spectrum of states. The wavelength, frequency and energy are equivalent measures in spectroscopy. One practical inconvenience of this is that numerous units used to characterize one and the same parameter – transition energy. The most frequently used values are collected in **Table 1.2. **

**Table 1.2 **

**Energy, frequency and wavelength units used in spectroscopy. **

The first three rows in **, it is conveniently used when energy or frequency dependence is presented. The last line in the table relates the photon energy to the energy of the electron in electric field. This appears to be a useful presentation as the effect of a photon absorption or emission is a transition of an electron from one energy level (orbital) to another, and the electron access energy can be used in some other reaction, e. g. in electron transfer from one molecule to another. **

In thermodynamic equilibrium any body absorbing light must emit equal amount of energy. This means that any body at temperature greater than absolute zero emits energy by electromagnetic radiation. The explanation of the spectrum of such thermal radiation was one of the fundamental discoveries in physics a century ago. To find the radiation density in a thermodynamically equilibrated system Max Planck proposed a quantum approach to the problem. He postulated that each oscillation mode of a closed cavity (resonator) could only take certain quantized energy**⁷ **

**(1.19) **

where *n *= 1, 2, 3 …. The probability to find energy *En *. Using **eq. (1.19) and Boltzmann statistics, Planck has shown that the spectral density of the radiation is **

**(1.20) **

where *h *is the Planck constant, *k *is the Boltzmann constant, *c *is the velocity of the light in vacuum, *v *is the frequency and *T *is the temperature. **Equation (1.20) describes emission of so-called black body, a body which has no any specific emission/absorption features and whose emission properties are completely determined by the thermodynamics, i. e. by eq. (1.20). **

The black body can serve as an emission model of a metal surface at relatively high temperature, e. g. of tungsten halogen lamp. For quantitative characterization of the radiation density a gray

coefficient is used, which is the ratio of the real body emission density to that of the ideal one. Usually the coefficient is less but close to 1. For metal surfaces the coefficient is a constant in a wide wavelength range.

In order to evaluate the emission properties of black bodies the spectral emittance can be used. It is defined as the total power emitted per unit wavelength interval into a solid angle 2π by an unit area of the black body, and is given by

**(1.21) **

where *c*1 = 2*πhc*² = 3.74 · 10−16 W·m² and *c*2 = *ch*/*kB *= 1.44 · 10−2 m·K. The spectral emittances calculated for the wavelength measured in nm and emitting area in cm² are presented in **Fig. 1.4. **

**Figure 1.4 **Black body spectral emittance calculated at temperatures 3000, 4000 and 5000 K according to **eq. (1.21). **

The maximum of the black body emission spectrum is (Wien’s law)

**(1.22) **

where the wavelength (λ*max*) is measured in nm and temperature (*T*) in Kelvins (K). Thus, the light source must have temperature of about 5000 °C to have emission maximum at the middle of the visible spectrum (500–550 nm). The total power emitted by unit area in a solid angle 2π is given by Stefan-Boltzmann law

**(1.23) **

where σ = 5.67 · 10−8 W·m−2K−4 is the Stefan-Boltzmann constant.

In lamps specification one can find two parameters which determine the emission intensity and the spectrum shape: color temperature and emissivity. The emissivity is the ratio of radiation emitted by the lamp to that of the ideal black body. The color temperature is such temperature of the ideal black body at which its spectrum (coloration), as given by **eq. (1.21), is similar to the emission spectrum of the lamp.⁸ **

The color temperature of tungsten halogen lamps ranges from 2000 to 3200 K, and typical emissivity is ∼ 0.4. If a lamp works at filament temperature 3000 K, its spectral emittance is *M *≈ 0.4 × 0.16 W·cm−2nm−1 ≈ 0.07 W·cm−2nm−1 at 600 nm. For an emitting filament area of 0.1 cm² the total power emitted at 600 nm in 1 nm wavelength range is 7 mW. The total power emitted by a tungsten lamp with filament size of 0.1 cm² at temperature 3000 K is *P *≈ 50 W.

The emission maximum of the lamp is *λmax *≈ 970 mn, so at shorter wavelengths the spectrum density is lower, e. g. 3 mW·nm−1 at 500 nm and 1 mW·nm−1 at 400 nm, and at longer is higher, e. g. 9 mW·nm−1 at 700 nm.

A typical color temperature of the cathode area of the electric arc is *T *≈ 6000 K. The bright cathode area diameter is usually 1–2 mm, so the surface of the emitting cathode sphere

can be estimated to be ≈ 3 mm² (1 mm sphere), which gives total power of *P *≈ 200 W. The emission maximum of such arc is at λ*max *≈ 480 nm. The spectrum density of the emission is 0.3 W·nm−1 at 500 nm (close to the maximum). At shorter wavelengths the spectrum densities are 0.27 and 0.17 W·nm−1 at 400 and 300 nm, respectively. At longer wavelengths the densities are 0.27 and 0.22 W·nm−1 at 600 and 700 nm, respectively. It should be noted, however, that this is an estimation of the plasma thermal emission only. Real spectra of Xe lamps consist of a number of relatively sharp emission lines on top of rather smooth black body like emission spectrum. This is due to excited states of Xe atom and its ions.

The black body

theory considers infinite number of energy states (oscillation modes) which are very close to each other (forming continuous spectrum). This approach ignores any individual properties of molecules or atoms probably involved in the emission or absorption process. In other words, this theory cannot be applied to gases or diluted dye solutions, single molecules and atoms have individual energy levels, which are well separated from each other. In the most simplified case one can consider a molecule with only two states *M*1 and *M*2 with energies *E*1 and *E*2, respectively:

This molecule will only interact with photons having energy *hv *= *E*2 – *E*1. The possible photo-reactions of the system are

1. photon absorption: *M*1 + *hv *→ *M*2;

2. thermal relaxation: *M*2 → *M*1 + Δ*E; *

3. spontaneous photon emission: *M*2 → *M*1 + *hv; *

4. stimulated photon emission: *M*2 + *hv *→ *M*1 + 2*hv*.

There are two types of reactions: spontaneous and stimulated. The stimulated reactions require a photon to occur, these are reactions (1) and (4). The spontaneous reactions do not require any external force to occur. They take place because of access energy accumulated by the system (e. g. a molecule in excited state), and they result in relaxation to a lower energy state by emitting a photon (reaction (3)) or releasing the access energy by some other means, e. g. thermal relaxation, reaction (3).

The transition probabilities can be expressed using Einstein’s coefficients *A *and *B. *Coefficient *A *describes spontaneous relaxation. In particular, for reaction (3), i. e. transition from state 2 to state 1,

**(1.25) **

where *N*2 is the population of state *M*2. Coefficients *B*21 and *B*12 describe stimulated absorption, reaction (1), and stimulated emission, reaction (4), respectively. Note that absorption is always stimulated.

In case of a narrow absorption and a broad stimulating radiation field, the kinetic equation for reaction (4) is

**(1.26) **

where ρ is the energy density.

Einstein has shown that

**(1.27) **

and

**(1.28) **

Thus, a single coefficient, for example *A*21, describes behavior of the two level system.

The relation (**1.27) deserves a separate comment as it provides a very general conclusion: the probability of stimulated emission is equal to the probability of absorption. This is of practical importance for laser applications, as will be discussed in Section 1.3. **

Real atoms and molecules have many energy levels and not all transitions between the levels are allowed. For instance, depending on spin multiplicity the states are divided on singlet states, having total spin quantum number 0, and triplet states with non-zero total spin. Transitions between triplet and singlet states are forbidden according to the spin conservation law. In practice this means that the rate of such transition, or the transition probability, is very low. The process in which the state of a molecule is changed from singlet to triplet state or backward is called inter-system crossing, and is one of the subjects of organic

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