The Laser Diode as a Light Source for Atomic Absorption Spectroscopy

Bryan Holmes Chem 226 Spring 2003

Laser Diode Atomic Absorption Spectroscopy

Introduction The development of laser diodes has increased the variety of practical applications for lasers as light sources in analytical spectroscopy. Lasers are excellent radiation sources due to their monochromaticity, good beam focusing capabilities, and small line-widths but have traditionally been expensive and difficult to operate, limiting their applications to specialized fields. Because the stability of the light source for atomic absorption spectroscopy is critical, tunable dye lasers and optical parametric oscillators are not well suited despite their high power densities. Due to their low cost, tunability of several nanometers, stable output power, compactness and ease of operation, diode lasers are finding an increasing number of uses in analytical spectroscopy. Some of these applications include laser-enhanced ionization spectroscopy, laser-excited atomic fluorescence spectroscopy, resonance ionization mass spectrometry, and specifically, atomic absorption spectroscopy. Diode laser technology is based on the optical emission of semi-conducting materials when population inversions are created with applied electrical potentials. The emission wavelength is mainly dependent on the semi-conducting material (Table 1). Diode lasers used in analytical spectroscopy are typically small (300 m x 300 m x 150 m), have low power (mW), and convert electrical power to produce optical radiation with high (10-30%) efficiency. Currently, diode lasers used for analytical spectroscopy are commercially available in the 630-1600 nm range. Second harmonic generation using non-linear crystals can yield 0.1 W between 335 and 410 nm and as much as 1-3 W in the 410-430 nm range. Due to poor conversion efficiencies, power losses are significant

3 with harmonic generation. Elemental transitions with energies matching ultraviolet photons energies will be accessible by sum frequency generation, a nonlinear photon conversion technique that will be able to yield ultra-violet radiation in the 10-100 nW range in the near future.[1] The limited wavelength ranges available with diode lasers, along with the poor conversion efficiencies of nonlinear optics limits the elements accessible by diode laser atomic absorption spectroscopy. Devices emitting in the yellow, green and blue regions have been developed but have had problems with short lifetimes (sometimes minutes or hours). Robust diode lasers emitting in the blue region are becoming commercially available.[2] For example, Melles Griot has recently introduced a violet diode laser with output at 405 nm and 440nm, producing 6 mW and 1 mW of power, respectively. Laser Diode Operation and Characteristics When atoms combine to form a solid (in this case, the semi-conducting material in a diode laser), the previously distinct energy levels of the free atoms overlap and broaden due to neighboring interactions. These broadened energy levels are called energy bands. The outermost electron's energy states broaden and combine to form the valence band. The collection of the lowest, unoccupied states forms the conduction band. The emission wavelengths () depend on the band gap of the semiconductor material. The energy spacing between the top of the highest filled energy band (the valence band) and the bottom of the lowest unfilled band (conduction band) is referred to as the band gap. Laser light in a diode is created by sending a current (injection current) through the active part of the diode containing the n- and p-type cladding layers (Figure 1) producing electrons (n-type, negative) and holes (p-type, positive) which

4 recombine to emit radiation. The most common type of diode laser confines this active area to a stripe generally 1-10 m wide, known as a stripe-geometry type laser. A narrow channel that confines the light in the active region controls the lasers spatial mode. Two types of diode lasers, gain guided or index guided, arise from different manipulations of the transverse laser mode. The confinement of the transverse laser mode is obtained by spatial variation of injection current density by resistive materials (gain guided) or the spatial confinement of light caused by changes in the materials index of refraction used in the lasers construction (index guided).[3] In both cases, limiting laser action to a narrow space forms a wave-guide where there is gain. Also, this manipulation improves the beam quality by helping to limit oscillations to a single transverse mode and often to a single longitudinal mode. [4] Gain-guided lasers are simpler than index guided lasers to make but rarely operate with stability in a single longitudinal mode due to a relatively weak wave-guiding effect. Also, non-linearity occurs in the output-versus-current characteristics. The advantage of gain-guided laser diodes is their ability to achieve higher powers. [4] For gain-guided lasers, powers of 200 mW are common and, in isolated cases, lasers up to 10 watts are commercially available. Most commercial single-stripe geometry, index-guided laser diodes emit radiation with a single spatial and longitudinal mode (Figure 2). However, side modes appear just above the lasing threshold at low injection currents. The cavity-length of a diode laser is several hundred micrometers and the longitudinal modes are separated by 0.1-1 nm, making it possible to measure these modes with a simple, low-resolution monochromator. Index-guided lasers are advantageous due to their narrow bandwidth, relatively stable

5 operation, and single longitudinal mode operation. Due to their design, they operate at a lower power than gain-guided lasers (typically 3-40 mW). [3] The wavelength emitted by a diode laser is primarily controlled by the band gap of the semi-conducting material. A modest tuning range is available by varying the current or temperature of the device. Generally, the tuning range is wide ( 30 nm) but tuning of the longitudinal modes is restricted to a few nanometers. [5] Modifying the temperature changes the laser cavity length as well as affects the gain within the lasing cavity. Gain and optical path-length have different dependencies on temperature. As seen in figure 3, the temperature-tuning curve exists, in the ideal case, as a staircase with sloping steps. The slope of the steps is caused by cavity mode tuning, where the jumps between steps are due to transitions from one longitudinal mode to another, a process known as mode-hopping ( 0.3 nm). Since diode lasers are susceptible to optical feedback (e.g. reflections), methods utilizing this sensitivity have been developed which use optical feedback to control unwanted wavelength changes, including mode-hopping. [3] Wavelength tuning is also accomplished by altering the injection current. Figure 4 shows the dependence of laser power on current and temperature. Changing the injection current affects the temperature in the cavity of the diode laser as well as the gain properties within the cavity, both varying the emitted wavelength. Mode-hopping is affected by both the injection current and temperature. [3] Compared to other tunable laser systems, restricted wavelength range and mode-hopping are limitations to this diode lasers as a light source. Many spectroscopists are optimistic that the fast growth of the semi-conductor industry as well as improvements in frequency-doubling technology will reduce these limitations. [6]

6 Analysis Techniques using DL-AAS Laser diodes have excellent spectroscopic and operational properties for analytical spectroscopy, providing an almost ideal line source for atomic absorption spectrometry. Diode lasers typically have line-widths of 20 MHz, which is about 30 times less than the atomic absorption line-widths for low pressure, room temperature atomic reservoirs. Compared to atomic line-widths in flames, diode lasers line-widths are about 100 times less. The stability, tunability and line characteristics of diode lasers increase experimental freedoms and opportunities within the field of atomic absorption spectrometry. [2] Generally, lasers offer advantages to the hollow cathode lamp as a light source for atomic absorption spectroscopy because they are monochromatic, directional, and have relatively high powers and narrow line-widths. The use of lasers adds simplicity to the experimental arrangement because monochromators are generally not needed for wavelength selection and simple photodiode detectors can be used to discriminate between the laser signal and background absorption. Hollow cathode lamps have a limited dynamic range of two to three orders of magnitude due to optical thickness in the centers of the absorption lines. For optically thin conditions, diode lasers tunability offers the advantage of selecting wavelengths on the wings of the absorption lines, extending the linear dynamic range many orders of magnitude for high concentrations of analyte. In addition, the linear dynamic range can be increased to lower concentrations by passing the laser radiation through a multi-pass cell. This is an absorption arrangement where radiation passes through the sample multiple times, usually using prisms or partially

7 reflecting mirrors. Also, tuning the diode laser far off the resonance transition, background corrections can be made for isobaric interferences and light scattering. [7] Experimental Setup A basic setup for diode laser atomic absorption spectroscopy using flame atomization is shown in figure 5. James Winefordner and coworkers [8] created this arrangement to measure rubidium using the 780.0 nm transition with a single-mode diode laser. Due to the diode lasers single mode operation, a wavelength-selection monochromators is not needed, which simplifies the experimental arrangement. The transmission signal was monitored using a diode array spectrometer and a monochromator/photomultiplier tube detector. Due to the diode array spectrometers low resolution and stray light problems, wavelength monitoring was performed with the monochromator/photomultiplier tube detector. This allowed for observing experimental figures of merit such as determining laser power parameters where 100% absorption occurred for the system. Background Corrections One of the valuable features of the laser diode is the availability of different wavelengths either by temperature/current tuning or through multi-mode operation. Despite the good stability of diode lasers, absorption experiments using this source are generally source noise limited. In order to improve sensitivity, tunable properties of the diode laser are utilized in background correction methods, which are ubiquitous techniques used in most modern diode laser atomic absorption spectroscopy. One method for background correction uses an off-resonance longitudinal mode of a multi-mode diode laser to monitor source noise at the detector. James Winefordner

8 and coworkers report the detection of lithium at its 670.780 nanometer resonance line using a diode-array spectrometer to monitor the multi-mode laser emission. Figure 6 shows the spectral lines of the multi-mode diode laser with the A line, representing the lithium resonance line position. For this experiment, the peaks line positions are generally stable, but the line intensities varied. As shown in figure 7, a peak-ratio measurement was performed using the resonance line (A) and a nearby off-resonance line (B), improving precision by correcting for source noise. Without this correction method, the absolute measurement of the absorbance line was 7% relative standard deviations for five trials. Precision was enhanced by the peak ratio method as seen in the improvements of the relative standard deviation to less than 1%. The sensitivity (1% absorption) was 20 ng/mL (ppb) and the detection limit (2 of blank) was 4 ppb. These values are similar to those (16 and 1 ppb, respectively) of conventional flame atomic absorption using a hollow cathode lamp. [6] Using a multimode laser diode, background correction is performed through peak absorption measurements similar to that of the Zeeman background correction using a hollow cathode lamp. To perform this correction, the multimode laser diode is tuned so that lines are positioned on both sides of the analyte resonance line. With multimode diode lasers, these nearby non-resonance lines are always accessible, allowing real-time background correction. This technique is especially beneficial to fast atom reservoirs such as the graphite furnace. [6] Due to its fast tuning capability, a background correction method known as wavelength modulation (WM) has been applied to laser diode atomic absorption spectroscopy. This method is most often used in atomic spectroscopy, where a small

9 wavelength range () is scanned rapidly back and forth across a wavelength interval, which is on and off the resonance line. Conventionally, measurements using wavelength modulations are performed by oscillation of an optical component (e.g. grating, interference filter, refractor plate) near the exit slit of a monochromator. The detector measures a signal whose magnitude is related to the signal change over the . [9] The signal to noise ratio depends on the spectral width of the source and on the spectral radiance. Laser diodes are better sources for wavelength modulation absorption spectroscopy compared to classical instrumentation because of their inherent simplicity, relatively narrow line-widths and fast tunability. Many different variations of wavelength modulation atomic absorption spectroscopy have been applied to address background correction issues due to dark detector noise, shot noise from the laser and photo-detection, laser excess noise, nonspecific absorption and optical excess noise. Liger and coworkers [10] report a doublemodulation and double-beam technique for the detection of gaseous meta-stable chlorine atoms to suppress these noise sources. In the single-beam, double modulation technique, the laser diode wavelength is altered sinusoidally at a specific frequency while the lowpressure microwave induced plasma or d.c. plasma atomization source is modulated at 5 kHz (Figure 8). The modulation frequency was mixed with the second harmonic of the laser modulation frequency and the signal for lock-in amplification. This is known as 2f wavelength modulation. The detection limit measured with a time constant of =1 is about 1x10-6 absorbance units. A double-beam arrangement with logarithmic differential amplification of the signals from the absorption and reference channels realized a detection limit of 2x10-7 absorbance units with a time constant of =1. Generally, hollow

10 cathode lamps express detection limits of 10-4 absorbance units. Diode lasers offer advantages to other sources due to their increased stability and fast tunability, reducing the detection limit by 2-3 orders of magnitude compared with classical atomic absorption spectroscopy. Isotope Selective Analysis When isotopic line shifts are sufficient to permit spectral separation, diode lasers offer the capability for isotope selective analysis. Sufficiently large isotopic shifts are only expressed for light (e.g, Li) and heavy (e.g., U) elements, when isotope shifts are larger than the Doppler broadening of the spectral lines. Isotope analysis is usually measured using mass spectrometry. The narrow line-widths emitted from single-mode diode lasers (typically 0.4 pm in red and infrared) offers capability for high-resolution absorption spectroscopy, making this a competitive technique with mass spectrometry for isotope analysis. Lead isotopes in aqueous samples were measured by Wizemann and Niemax[11] using atomic absorption in a furnace atomic non-thermal excitation spectroscopy atomizer (FANES) which is an electrothermally heated graphite tube atomizer with integrated low-pressure d.c. discharge (Figure 9). A frequency-doubled diode laser delivers radiation at 405.78 nm as the excitation source and calibration was made by isotope dilution. In order to achieve isotopic spectral resolution, measurements were made in a low-pressure atom reservoir to limit pressure broadening and the FANES instrument populated the 6p2 3P2 meta-stable electronic level. As the FANES is prone to severe matrix effects, application of wavelength modulation and isotope dilution provided a matrix independent analysis. Detection limits are measured to be 4.5x10-3

11 absorbance units for 206Pb and 208Pb. These are relatively poor values. These measurements are shot-noise limited due to the low laser power of the frequency-doubled radiation but should theoretically increase by at least 3 orders of magnitude by using laser powers of 2 milliwatts instead of 20 nanowatts. It is expected that new types of laser diodes with higher powers (milliwatts) and operational wavelengths in the blue will help realize these goals. Isotope selectivity using diode laser absorption techniques has also been shown with solid samples. Recently, the coupling of laser ablation with diode laser-based technique has proved to be useful for probing gaseous atoms from solid samples in the expanding plasma plume. Quentmeier and coworkers [12] have demonstrated this technique for detection of the 235U and 238U isotopes by laser ablation diode laser atomic absorption spectroscopy. The need for a field instrument that is compact and portable led to the development of many different optical techniques for solids in order to minimize instrumental contamination. Because of the high concentrations of uranium atoms in the laser plume, absorption proved to be a superior technique, even compared to fluorescence. Unfortunately, due to spatial variations in the laser plume, initial studies on these methods suffered from poor precision. Niemax and coworkers [13] improved the sensitivity and selectivity of laser ablation diode laser atomic absorption spectroscopy by characterizing the spatial and temporal characteristics of the laser plume. This was done by using two diode lasers as shown in figure 10 to measure the absorbance of the Uranium isotopes simultaneously in order to improve reproducibility and accuracy. A relative standard deviation of 2-3% is a significant improvement using this double-beam technique. This method is a competitive

12 to alternative techniques such as laser-induced plasma emission and optogalvanic spectroscopy. Multi-element Analysis The laser diodes simplicity, small size and low-cost, along with its many other virtues, makes it a good source for multi-elemental analysis. The well-collimated beam of a single diode laser allows a setup where many diode lasers can be directed simultaneously through an absorption pathlength. Hergenroder and Niemax [14] demonstrated multi-element detection of rubidium and barium in a graphite furnace. Two semi-conductor lasers were tuned to the rubidium resonance line 5s 2S1/2-5p 2P3/2 at 780.2 nm and the barium inter-combination line 6s2 1S0-6s6p3P1 at 791.1 nm, respectively. For background correction, the wavelength modulation technique was used. As the power of the diode laser is modulated sinusoidally with a frequency, f1, Fourier analysis is used to analyze the signal measured with a simple photodiode (Figure 11). By applying two separate diode lasers at different modulation frequencies, the information can be converted from the time to the frequency domain, where the relevant components of the Fourier spectrum can be associated with specific elements. Groll and Niemax [15] extended this method to the simultaneous detection of lithium, potassium, rubidium, and cesium in an air-acetylene flame and of lithium, potassium, rubidium, cesium, strontium and barium in a commercial graphite-tube furnace using six independent laser diodes. Fourier analysis of more than two elements is possible but requires a higher quality and more expensive analyzer. Instead, the arrangement works as a quasi-simultaneous time-multiplex technique whereby only one of the diode lasers is tuned to the resonance position and all other wavelength are

13 positioned off-resonance. These measurements are comparable with the best data of conventional single-element atomic absorption spectroscopy using hollow cathode lamps published in the literature. The authors feel that optimization of the flame and graphite furnace will have a significant impact on lowering the detection limits for this technique even further. Present and Future Diode Laser Absorption Sensors Laser diode sensors are currently being developed for interrogation of highpressure, transient, and sometimes hostile environments. Many laser-based strategies use inefficient, complex, large and expensive pulsed lasers that are not practical for real-time control applications. Advanced, wavelength agile diode lasers, known as, vertical cavity surface-emitting lasers (VCSEL), are now available that offer rapid, broad wavelength scanning capabilities compared to traditional diode-laser absorption techniques. The VCSEL's principle of operation is similar to those of conventional index-guided, edgeemitting semiconductor lasers. The lasing cavity of the VCSEL is an electrically pumped gain region, also called the active region, which emits light. Layers of varying semiconductor materials above and below the gain region create mirrors. Each mirror reflects a narrow range of wavelengths back into the cavity causing light emission at a single wavelength. Broad-wavelength scanning diode lasers are increasing the analytical possibilities of absorption spectroscopy. Sanders and coworkers [16] are developing wavelength-agile sensing strategies for in situ detection of species in combustion sources. In this work, cesium seeding is used as a tracer species in order to characterize this strategy. A

14 VCSEL is used to monitor the temperature and pressure in a pulse detonation engine. By using aggressive injection current modulation, the VCSEL is scanned through a 10 cm-1 spectral range at MHz rates. This is about ten times the scanning range and 1000 times the scanning rate of a conventional diode laser. The VCSEL scans 10 cm-1 every microsecond to measure cesium absorption and thermal emission waveforms (Figure 12). This is significant because absorption bands in the high pressure and high temperature gases can increase the collisional widths up to 4 cm-1. The fast, broad-scanning capabilities of the VCSEL allows for measurements in these harsh conditions. The advancement of diode laser technology will make shorter wavelengths available so that real-time detection of native species in combustion gases is increasingly practical. The fast timeframes of the relatively cheap and simple VCSEL allows for the rapid acquisition of data in dynamic and hostile environments, such as the high temperature and pressure flue gases. Improvements for diode-laser sensor systems will include faster algorithms for reducing the computational time, incorporation of laser-diode multiplexing techniques for simultaneous multi-species measurements, and refined digital filtering techniques for detection. Oscillator strength (fabs) and Atomic Absorption Spectroscopy The oscillator strength is a fundamental quantity in analytical atomic spectroscopy. It determines the sensitivity of a transition and is described by the number of electrons per atom or molecule undergoing a radiative transition. This value must be accurately known if one is to correlate an absorption signal to the elemental concentration. Hannaford [17] describes a surprising result related to the oscillator strength, namely an independence of the peak absorption coefficient on the absorption

15 oscillator strength for atoms where the absorption line is broadened only by natural broadening. The peak absorption only becomes dependant on the absorption oscillator strength when broadening processes such as Doppler and collisional broadening occur. Because of this relationship, a spectral line, which has a natural width of 10-4 will have a peak absorption 100 times larger than an absorption line which has a collisional width of 10-2 . As Doppler and collisional broadening become more significant, absorbance becomes increasingly dependant on the oscillator strength, fabs, and the halfwidths of the Doppler broadened and collisional broadened absorption lines. [2] In order to interrogate an atomic absorption line where the broadening is only due to natural broadening, a radiation source would have to have an emission line half-width less than the natural broadened absorption line. Diode lasers are a suitable source for an application of this nature, in which atoms are cooled so that all broadening sources except for natural broadening are minimized. The limitations and challenge to these investigations is that the cooling of atoms is complex, expensive, and sometimes impossible with many atomic absorption atomizers. [2] Hannaford and McLean [18] have shown theoretically that the absorption signal at the line center for ultra-cold, quasi-stationary absorbing atoms and a nearmonochromatic resonant radiation source is independent of the oscillator strength. The narrow lines widths of a diode laser are used to measure these relationships with cesium atoms. The goal is to perform oscillator strength-free atomic absorption where the atomic absorption signal at the line center is equal in magnitude for both strong and weak (closed) transitions of the same wavelength. Hannaford and McLean demonstrate that it is possible to perform atomic absorption experiments based on ultra-cold, laser-cooled

16 cesium atoms using a diode laser source. These experimental conditions approximate the conditions necessary for oscillator-strength free atomic absorption but major technical challenges must be addressed before the analysis of ultra-cold atoms with diode lasers can be used in atomic analytical spectroscopy. Conclusions Diode lasers offer many benefits compared to conventional sources for atomic absorption spectroscopy. Their stability, low-cost, simplicity, narrow line-widths, and rapid tunability make diode lasers an advantageous source compared to the traditional, hollow cathode lamp. Because of the limited wavelengths available with diode lasers, roughly 45% of the elements measured by hollow cathode sources are accessible. Detection limits for basic experimental arrangements using diode lasers are on the same order as the detection limits for hollow cathode lamp atomic absorption spectroscopy. By applying background correction techniques using the diode lasers rapid scanning abilities, increased selectivity, a larger linear dynamic range and much lower detection limits can be achieved. Due to market driven forces, diode lasers with shorter wavelengths and higher powers are becoming more available, increasing the number of elements that are measurable by diode laser atomic absorption spectroscopy.

17 Literature Cited 1. Nakamura, S., Senoh, M., Nagahama, N., InGaN/GaN/AlGaN-based laser diodes with cleaved facets grown on GaN substrates. Applied Physical Letters, 1998. 72: p. 211. 2. Winefordner, J.D., et al., Novel uses of lasers in atomic spectroscopy. Journal of Analytical Atomic Spectrometry, 2000. 15(9): p. 1161-1189. 3. Wieman, C.E., Hollberg, L., Using diode lasers for atomic physics. Review of Scientific Instruments, 1991. 62: p. 1-20. 4. 5. 6. Hecht, J., The Laser Guidebook. 2 ed. 1992, New York: McGraw-Hill Inc. Imasaka, T., Diode lasers in analytical chemistry. Talanta, 1999. 48: p. 305-320. Ng, K.C., et al., The applicability of a multiple-mode diode laser in flame atomic absorption spectroscopy. Applied Spectroscopy, 1990. 44(5): p. 849-52. 7. Niemax, K., H. Groll, and C. Schnuerer-Patschan, Element analysis by diode laser spectroscopy. Spectrochimica Acta Reviews, 1993. 15(5): p. 349-77. 8. Ng, K.C., et al., Flame atomic absorption spectroscopy using a single-mode laser diode as the line source. Applied Spectroscopy, 1990. 44(6): p. 1094-6. 9. Ingle, J.D., Crouch S. R., Spectrochemical Analysis. 1988, Englewood Cliffs: Prentice Hall Inc. 445-446. 10. Liger, V., et al., Diode-laser atomic-absorption spectrometry by the double-beamdouble-modulation technique. Spectrochimica Acta, Part B: Atomic Spectroscopy, 1997. 52B(8): p. 1125-1138.

18 11. Wizemann, H.D. and K. Niemax, Cancellation of Matrix Effects and Calibration by Isotope Dilution in Isotope-Selective Diode Laser Atomic Absorption Spectrometry. Analytical Chemistry, 1997. 69(20): p. 4291-4293. 12. Quentmeier, A., Bolshov, M, and Niemax, K., Measurement of uranium isotope ratios in solid samples using laser ablation and diode laser-excited atomic absorption spectrometry. Spectrochimica Acta, Part B: Atomic Spectroscopy, 2001. 56: p. 45-55. 13. Liu, H., A. Quentmeier, and K. Niemax, Diode laser absorption measurement of uranium isotope ratios in solid samples using laser ablation. Spectrochimica Acta, Part B: Atomic Spectroscopy, 2002. 57B(10): p. 1611-1623. 14. Hergenroeder, R. and K. Niemax, Laser atomic absorption spectroscopy applying semiconductor diode lasers. Spectrochimica Acta, Part B: Atomic Spectroscopy, 1988. 43B(12): p. 1443-9. 15. Groll, H., K. Niemax, Multielement diode laser atomic absorption spectrometry in graphite tube furnaces and analytical flames. Spectrochimica Acta, 1993. 48B(5): p. 633-641. 16. Sanders, S.T., Daniel W. Mattison, Lin Ma, Jay B. Jeffries, and Ronald K. Hanson, Wavelength-agile diode laser sensing strategies for monitoring gas properties in optically harsh flows: application in cesium-seeded pulse detonation engine. Optics Express, 2002. 10(12): p. 505-514. 17. Hannaford, P., The oscillator strength in atomic absorption spectroscopy. Spectrochimica Acta, Part B: Atomic Spectroscopy, 1994. 49B(12-14): p. 15811593.

19 18. Hannaford, P. and R.J. McLean, Atomic absorption with ultracold atoms. Spectrochimica Acta, Part B: Atomic Spectroscopy, 1999. 54B(14): p. 2183-2194.

Tables and Figures Table 1

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Figure 1

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Figure 2

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Figure 3

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Figure 4

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Figure 5

25 Figure 6

26 Figure 7

27 Figure 8

Figure 9

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Figure 10

Figure 11

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Figure 12