Experimental determination of the effective point of measurement of cylindrical ionization chambers for high-energy photon and electron beams

Measurements of depth-dose curves in water phantom using a cylindrical ionization chamber require that its effective point of measurement is located at the measuring depth. Recommendations for the position of the effective point of measurement with respect to the central axis valid for high-energy electron and photon beams are given in dosimetry protocols. According to these protocols, the use of a constant shift P_eff is currently recommended. However, this is still based on a very limited set of experimental results. It is therefore expected that an improved knowledge of the exact position of the effective point of measurement will further improve the accuracy of dosimetry. Recent publications have revealed that the position of the effective point of measurement is indeed varying with beam energy, field size and also with chamber geometry. The aim of this study is to investigate whether the shift of P_eff can be taken to be constant and independent from the beam energy. An experimental determination of the effective point of measurement is presented based on a comparison between cylindrical chambers and a plane-parallel chamber using conventional dosimetry equipment. For electron beams, the determination is based on the comparison of halfvalue depth R₅₀ between the cylindrical chamber of interest and a well guarded plane-parallel Roos chamber. For photon beams, the depth of dose maximum, d_max, the depth of 80% dose, d₈₀, and the dose parameter PDD(10) were used. It was again found that the effective point of measurement for both, electron and photon beams Dosimetry, depends on the beam energy. The deviation from a constant value remains very small for photons, whereas significant deviations were found for electrons. It is therefore concluded that use of a single upstream shift value from the centre of the cylindrical chamber as recommended in current dosimetry protocols is adequate for photons, however inadequate for accurate electron beam dosimetry.     

Dosimetric impacts of beam-hardening filter removal for the CyberKnife system

PurposeEquipment refurbishment was performed to remove the beam-hardening filter (BHF) from the CyberKnife system (CK). This study aimed to confirm the change in the beam characteristics between the conventional CK (present-BHF CK) and CK after the BHF was removed (absent-BHF CK) and evaluate the impact of BHF removal on the beam quality correction factors k_Q.MethodsThe experimental measurements of the beam characteristics of the present- and absent-BHF CKs were compared. The CKs were modeled using Monte Carlo simulations (MCs). The energy fluence spectra were calculated using MCs. Finally, k_Q were estimated by combining the MC results and analytic calculations based on the TRS-398 and TRS-483 approaches.ResultsAll gamma values for percent depth doses and beam profiles between each CK were less than 0.5 following the 3%/1 mm criteria. The percentage differences for tissue-phantom ratios at depths of 20 and 10 cm and percentage depth doses at 10 cm between each CK were −1.20% and −0.97%, respectively. The MC results demonstrated that the photon energy fluence spectrum of the absent-BHF CK was softer than that of the present-BHF CK. The k_Q values for the absent-BHF CK were in agreement within 0.02% with those for the present-BHF CK.ConclusionsThe photon energy fluence spectrum was softened by the removal of BHF. However, no remarkable impact was observed for the measured beam characteristics and k_Q. Therefore, the previous findings of the k_Q values for the present-BHF CK can be directly used for the absent-BHF CK.     

Assessment of ionization chamber correction factors in photon beams using a time saving strategy with PENELOPE code

The purpose of this study was to investigate Monte Carlo-based perturbation and beam quality correction factors for ionization chambers in photon beams using a saving time strategy with PENELOPE code. Simulations for calculating absorbed doses to water using full spectra of photon beams impinging the whole water phantom and those using a phase-space file previously stored around the point of interest were performed and compared. The widely used NE2571 ionization chamber was modeled with PENELOPE using data from the literature in order to calculate absorbed doses to the air cavity of the chamber. Absorbed doses to water at reference depth were also calculated for providing the perturbation and beam quality correction factors for that chamber in high energy photon beams. Results obtained in this study show that simulations with phase-space files appropriately stored can be up to ten times shorter than using a full spectrum of photon beams in the input-file. Values of k_Q and its components for the NE2571 ionization chamber showed good agreement with published values in the literature and are provided with typical statistical uncertainties of 0.2%. Comparisons to k_Q values published in current dosimetry protocols such as the AAPM TG-51 and IAEA TRS-398 showed maximum percentage differences of 0.1% and 0.6% respectively. The proposed strategy presented a significant efficiency gain and can be applied for a variety of ionization chambers and clinical photon beams.     

Design of commissioning process for Halcyon™ linac with a new rigid board: A clinical experience

PurposeThis study performed the accurate measurements of beam profiles with a new rigid board, which was consistent with the supplied reference beam profiles (RBPs) for clinical Halcyon model.MethodsPercentage depth doses (PDDs), lateral and diagonal dose profiles were measured and compared with RBPs. A water tank was set on the rigid board bridged Halcyon bore without sagging and source-to-surface distance was 90.0 cm. Field sizes were from 2.0 to 28.0 cm squares and depths of lateral and diagonal dose profiles were 1.3, 5.0, 10.0, and 20.0 cm. For the PDD, the depth of maximum dose (d_max), PDD value at depth of 10.0 cm (PDD₁₀), and absolute dose difference (DD) between RBP and measured beam profiles (MBP) were evaluated. For lateral and diagonal dose profiles, DDs for the whole and divided areas (central, shoulder, and extended areas) defined by third derivative, and distance-to-agreement (DTA) in the penumbra area were evaluated.ResultsFor PDDs, the differences of d_max and PDD₁₀ and DD beyond the d_max were within 1.0 mm, 0.3%, and 1.0%, respectively. For lateral and diagonal dose profiles, the DDs reached approximately 5.0% in the whole area because of penumbra area, while the DDs in the central, shoulder, and extended areas were within 1.0%, 2.0%, and 1.0%, respectively. The DTAs in the penumbra area were within 0.8 mm.ConclusionsThe supplied RBPs can be used clinically owing to the good agreement with the accurate MBPs with rigid board.     

High throughput film dosimetry in homogeneous and heterogeneous media for a small animal irradiator

PurposeWe have established a high-throughput Gafchromic film dosimetry protocol for narrow kilovoltage beams in homogeneous and heterogeneous media for small-animal radiotherapy applications. The kV beam characterization is based on extensive Gafchromic film dosimetry data acquired in homogeneous and heterogeneous media. An empirical model is used for parameterization of depth and off-axis dependence of measured data.MethodsWe have modified previously published methods of film dosimetry to suit the specific tasks of the study. Unlike film protocols used in previous studies, our protocol employs simultaneous multi-channel scanning and analysis of up to nine Gafchromic films per scan. A scanner and background correction were implemented to improve accuracy of the measurements. Measurements were taken in homogeneous and inhomogeneous phantoms at 220 kVp and a field size of 5 × 5 mm². The results were compared against Monte Carlo simulations.ResultsDose differences caused by variations in background signal were effectively removed by the corrections applied. Measurements in homogeneous phantoms were used to empirically characterize beam data in homogeneous and heterogeneous media. Film measurements in inhomogeneous phantoms and their empirical parameterization differed by about 2%–3%. The model differed from MC by about 1% (water, lung) to 7% (bone). Good agreement was found for measured and modelled off-axis ratios.ConclusionsEBT2 films are a valuable tool for characterization of narrow kV beams, though care must be taken to eliminate disturbances caused by varying background signals. The usefulness of the empirical beam model in interpretation and parameterization of film data was demonstrated.     

Calibration of MTT assay in proton beams using radiochromic films

PurposeThis study provides methodology of calibrating as well as controlling the output for an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) colorimetric assay irradiated in a low energy proton beam using EBT3-model GAFCHROMIC^TM film, without correcting for quenching effect.MethodsA calibrated Markus ionization chamber was used to measure the depth dose and beam output for 26.5 MeV protons produced by a CS30 cyclotron. A time-controlled aluminum cylinder was added in front of the horizontal beam-exit serving as a radiation shutter. Following the TRS-398 reference dosimetry protocol for proton beams, the output was calibrated in water at a reference depth of 3 mm. EBT3 film was calibrated for doses up to 8 Gy at the same depth. To verify the dose distribution for each 96-well MTT assay plate, EBT3 film was placed at the reference depth during irradiation and cell doses were scaled by measured percent depth dose (PDD) data.ResultsThe radiochromic film dosimetry system in this study provides dose measurements with an uncertainty better than 3.3% for doses higher than 1 Gy. From a single exposure and utilizing the Gaussian shape of the beam, multiple dose points can be obtained within different wells of the same plate ranging from 6.9 Gy (sigma ∼4%) in the central well, and 2 Gy (sigma ∼8%) for wells positioned closer to the periphery.ConclusionsWe described a methodology for radiochromic film-based dose monitoring system, using low-energy protons, which can be used for the MTT assay in any proton beam, except within Bragg peak region.     

Biofilm thickness measurements by light microscopy

Light microscopy has been used to measure biofilm thickness. The vertical displacement of the sample required to focus from the biofilm-liquid interface to the biofilm-substratum interface is measured by the stage micrometer. Biofilm thickness is proportional, but not equal, to the measured vertical displacement. An expression for the proportionality constant, k_f, in terms of refractive indices is determined from a geometric analysis of the light path. k_f can be estimated as the ratio of the refractive index of the film to the refractive index of the media interfacing the film between the objective lens and the sample. The thickness of any transparent film may be determined by light microscopy when the refractive index of the film is known.     

Impact of transverse magnetic fields on dose response of a nanoDot OSLD in megavoltage photon beams

PurposeWe investigated the impact of transverse magnetic fields on the dose response of a nanoDot optically stimulated luminescence dosimetry (OSLD) in megavoltage photon beams.MethodsThe nanoDot OSLD response was calculated via Monte Carlo (MC) simulations. The responses R_Q and R_Q,B without and with the transverse magnetic fields of 0.35–3 T were analyzed as a function of depth at a 10 cm × 10 cm field for 4–18 MV photons in a solid water phantom. All responses were determined based on comparisons with the response under the reference conditions (depth of 10 cm and a 10 cm × 10 cm field) for 6 MV without the magnetic field. In addition, the influence of air-gaps on the nanoDot response in the magnetic field was estimated according to Burlin’s general cavity theory.ResultsThe R_Q as a function of depth for 4–18 MV ranged from 1.013 to 0.993, excepting the buildup region. The R_Q,B increased from 2.8% to 1.5% at 1.5 T and decreased from 3.0% to 1.1% at 3 T in comparison with R_Q as the photon energy increased. The depth dependence of R_Q,B was less than 1%, excepting the buildup region. The top air-gap and the bottom air- gap were responsible for the response reduction and the response increase, respectively.ConclusionsThe response R_Q,B varied depending on the magnetic field intensity, and the variation of R_Q,B reduced as the photon beam energy increased. The air-gaps affected the dose deposition in the magnetic fields.     

Dosimetric evaluation of Gafchromic EBT2 film for breast intraoperative electron radiotherapy verification

PurposeQuality assurance (QA) is one of the most important issues that should be addressed for intraoperative electron radiotherapy (IOERT), which is not benefiting from image-based treatment planning system. The aim of this study is to evaluate the dosimetric characteristics of Gafchromic EBT2 film for breast IOERT QA procedure.MethodsDue to the fact that some dedicated accelerators are being used for IOERT, dependence of the film response to energy, field size, dose rate and incidence angle of electron beam from the LIAC IOERT accelerator was studied. Then, film response curve to breast IOERT doses was obtained and its accuracy was evaluated and justified through comparison to the results of ionometric dosimetry.ResultsThe results of this study indicated that there are no significant differences between the film responses at different energies of 6, 8, 10 and 12 MeV (P-value = 0.99). Similarly, no field size dependency was found when evaluating the response of the film to different field sizes ranging from 4 to 10 cm (P-value = 0.94). Film response was found to be independent of the dose rate of intraoperative electron beam (P-value = 0.12). Film response variations with changing the beam incidence angle were not significant (P-value > 0.8). Calibration curve at the dose range of 8–24 Gy had an acceptable accuracy. The difference between the results of film dosimetry and ionometric dosimetry was around 5% which was in agreement with the results of dose uncertainty estimation.ConclusionThe EBT2 film was found to be a potentially appropriate tool for breast IOERT verification.     

The use of non-standard CT conversion ramps for Monte Carlo verification of 6 MV prostate IMRT plans

Monte Carlo (MC) dose calculation algorithms have been widely used to verify the accuracy of intensity-modulated radiotherapy (IMRT) dose distributions computed by conventional algorithms due to the ability to precisely account for the effects of tissue inhomogeneities and multileaf collimator characteristics. Both algorithms present, however, a particular difference in terms of dose calculation and report. Whereas dose from conventional methods is traditionally computed and reported as the water-equivalent dose (D_w), MC dose algorithms calculate and report dose to medium (D_m). In order to compare consistently both methods, the conversion of MC D_m into D_w is therefore necessary.This study aims to assess the effect of applying the conversion of MC-based D_m distributions to D_w for prostate IMRT plans generated for 6 MV photon beams. MC phantoms were created from the patient CT images using three different ramps to convert CT numbers into material and mass density: a conventional four material ramp (CTCREATE) and two simplified CT conversion ramps: (1) air and water with variable densities and (2) air and water with unit density. MC simulations were performed using the BEAMnrc code for the treatment head simulation and the DOSXYZnrc code for the patient dose calculation. The conversion of D_m to D_w by scaling with the stopping power ratios of water to medium was also performed in a post-MC calculation process.The comparison of MC dose distributions calculated in conventional and simplified (water with variable densities) phantoms showed that the effect of material composition on dose-volume histograms (DVH) was less than 1% for soft tissue and about 2.5% near and inside bone structures. The effect of material density on DVH was less than 1% for all tissues through the comparison of MC distributions performed in the two simplified phantoms considering water. Additionally, MC dose distributions were compared with the predictions from an Eclipse treatment planning system (TPS), which employed a pencil beam convolution (PBC) algorithm with Modified Batho Power Law heterogeneity correction. Eclipse PBC and MC calculations (conventional and simplified phantoms) agreed well (<1%) for soft tissues. For femoral heads, differences up to 3% were observed between the DVH for Eclipse PBC and MC calculated in conventional phantoms. The use of the CT conversion ramp of water with variable densities for MC simulations showed no dose discrepancies (0.5%) with the PBC algorithm. Moreover, converting D_m to D_w using mass stopping power ratios resulted in a significant shift (up to 6%) in the DVH for the femoral heads compared to the Eclipse PBC one.Our results show that, for prostate IMRT plans delivered with 6 MV photon beams, no conversion of MC dose from medium to water using stopping power ratio is needed. In contrast, MC dose calculations using water with variable density may be a simple way to solve the problem found using the dose conversion method based on the stopping power ratio.     

Energy and dose dependence of GafChromic EBT3-V3 film across a wide energy range

AimTo determine the energy and dose dependence of GafChromic EBT3-V3 film over an energy range 0.2 mm Al HVL to 6 MV.BackgroundThe decay scheme of a brachytherapy source may be complex and the spectrum of energy can be wide. LiF TLDs are the golden standard recommended for dosimetric measures in brachytherapy, for their energy independence, but TLDs could be not available in some centres. An alternative way to perform dose measurements is to use GafChromic films, but they show energy dependence.Methods and materialsFilms have been irradiated at increasing dose with three different beams: 6 MV beam, TPR20, 10 = (0.684 ± 0.01), HVL = (2.00 ± 0.01)mmAl and HVL = (0.20 ± 0.01)mmAl. Calibration curves were generated using the same dose range (0cGy to 850cGy) for the three energies. Using the 6 MV calibration curve as reference, the film response in terms of net optical density (OD) was evaluated.ResultsThe difference in the calibration curve obtained by irradiating the film with 6 MV and 2 mm Al HVL energy beams is less than 3 %, within the calibration uncertainty, in the dose range 500-850cGy. The OD of EBT3-V3 film is significantly lower at 0.2 mmAl HVL compared to 6 MV, showing differences up to 25 %.ConclusionWithin the range 6 MV-2 mm Al HVL and dose higher than 500cGy, GafChromic EBT3-V3 films are energy independent. In this dose range, films can be calibrated in a simple geometry, using a 6 MV Linac beam, and can be used for brachytherapy sources dose measures. The use of EBT3 films can be extended to reference dosimetry in Ir-192 clinical brachytherapy.     

Dosimetric impact of failing to apply correction factors to ion recombination in percentage depth dose measurements and the volume-averaging effect in flattening filter-free beams

PurposeTo examine whether it is essential to apply correction factors for ion recombination (k_S) to percentage depth dose (PDD) measurements and to the volume-averaging effect (k_vol) to ensure accurate absolute dose calibration for flattening filter-free (FFF) beams for the most commonly used ionization chambers.MethodsWe surveyed medical physicists worldwide (n = 159) to identify the five most common ionization chamber combinations used for absolute and relative reference dosimetry of FFF beams. We then assessed the overall absolute dose calibration error for FFF beams of the Artiste Siemens and TrueBeam Varian linear accelerators resulting from failing to apply correction factors k_S in the PDD(10) and the volume-averaging effect (k_vol) to such chamber combinations.ResultsAll the chamber combinations examined—the Farmer PTW 30013 ionization chamber used for absolute dosimetry, and the PTW 31010, PTW 30013, IBA CC04, IBA CC13, and PTW 31021 ionization chambers used for PDD curves measurements—showed non-negligible errors (≥0.5%). The largest error (1.6%) was found for the combination of the Farmer PTW 30013 chamber with the IBA CC13 chamber, which was the most widely used chamber combination in our survey.ConclusionsBased on our findings, we strongly recommend assessing the impact of failing to apply correction factors k_S in the PDD(10) and k_vol prior to using any chamber type for FFF beam reference dosimetry purposes.     

Ion Transport through Monolayers and Interfacial Films

Ion transport through monolayers and through several molecules of thick films at the mercury/water interface is discussed. The permeability of the monolayer is described by a rate constant, k_c. The permeability of a thin but not monomolecular film is expressed as a function of the thickness of the film, the diffusion coefficient of the permeant in the film, and the distribution coefficient between the film and the bulk of the solution. The rate constant k_c is expressed in terms of absolute rate processes. In the absence of specific interactions, the activation energy is composed of three terms: (a) electrostatic interaction between the permeating ion and the charged monolayer, (b) monolayer compression work of forming a hole for passage of the ions, and (c) energy of boundary line formation between the monolayer and the hole. The contribution of the third term is especially marked in condensed monolayers. Ions are bound weakly to the monolayers of the dipolar ion lecithin, which complicates the transport problem in this system. The retardation of oxygen reduction by the lecithin monolayer is of particular interest.     

Skin models and their impact on mean glandular dose in mammography

PurposeTo quantify the influence of different skin models on mammographic breast dosimetry, based on dosimetric protocols and recent breast skin thickness findings.MethodsBy using an adapted PENELOPE (v. 2014) + PenEasy (v. 2015) Monte Carlo (MC) code, simulations were performed in order to obtain the mean glandular dose (MGD), the normalized MGD by incident air Kerma (DgN), and the glandular depth dose (GDD(z)). The geometry was based on a cranio-caudal mammographic examination. Monoenergetic and polyenergetic beams were implemented, for a breast thickness from 2 cm to 9 cm, with different compositions. Seven skin models were used: a 5 mm adipose layer; a skin layer ranging from 5 mm to 1.45 mm, a 1.45 mm skin thickness with a subcutaneous adipose layer of 2 mm and 3.55 mm.ResultsThe differences, for monoenergetic beams, are higher (up to 200%) for lower energies (8 keV), thicker and low glandular content breasts, decreasing to less than 5% at 40 keV. Without a skin layer, the differences reach a maximum of 1240%. The relative difference in DgN values for 1.45 mm skin and 5 mm adipose layers and polyenergetic beams varies from −14% to 12%.ConclusionsThe implemented MC code is suitable for mammography dosimetry calculations. The skin models have major impacts on MGD values, and the results complement previous literature findings. The current protocols should be updated to include a more realistic skin model, which provides a reliable breast dose estimation.     

Dosimetry of small photon fields in the presence of bone heterogeneity using MAGIC polymer gel,Gafchromic film,and Monte Carlo simulation

BackgroundThe presence of heterogeneity within the radiation field increases the challenges of small field dosimetry. In this study, the performance of MAGIC polymer gel was evaluated in the dosimetry of small fields beyond bone heterogeneity.Materials and methodsCircular field sizes of 5, 10, 20 and 30 mm were used and Polytetrafluoroethylene with density of 2.2 g/cm³ was used as the bone equivalent material. The PDD curves, beam profiles, and penumbra widths were measured using MAGIC polymer gel, EBT2 film, and Monte Carlo simulation.ResultsThe maximum differences between MAGIC and EBT2 are 6.1, 4.7, 2.4, and 2.2 for PDD curves at 5, 10, 20, and 30 mm circular fields, respectively. The dose differences and distance to agreement between MAGIC and MC were within 1.89%/0.46 mm, 1.66%/0.43 mm, 1.28%/0.77 mm, and 1.31%/0.81 mm for beam profile values behind bone heterogeneity at 5, 10, 20, and 30 mm field sizes, respectively.ConclusionThe results presented that the MAGIC polymer gel dosimeter is a proper instrument for dosimetry beyond high density heterogeneity.     

Effect of ventilation on the production of acetaldehyde by Zymomonas mobilis

The production of acetaldehyde, a flavoring agent in food, by Zymomonas mobilis was carried out in batch culture. The volatilization rate constant (k_v) of acetaldehyde and the initial volumetric oxygen transfer coefficient (k_La⁰) in an Erlenmeyer flask with a cotton-plug (cotton-flask) and an aerated-flask with a forced-air system (aerated-flask) were measured. The culture environment in the aerated-flask was found to be very different from that in the cotton-flask. Cell growth in a cotton-flask was strongly inhibited, making practical acetaldehyde production in cotton-flask very difficult. On the other hand, acetaldehyde production in the aerated-flask increased while the fermentation time decreased with increases in the air flow rate. The k_v value of acetaldehyde in a jar fermentor was affected mainly by air flow rate. By considering both the oxygen transfer rate and the ventilation effect on the culture, it was possible to scale-up from the aerated-flask to a jar fermentor. In the jar fermentor, production of acetaldehyde and growth inhibition by acetaldehyde were affected mainly by the k_La⁰ and k_v, values, respectively. The overall production of acetaldehyde in the jar fermentor under the optimum k_La⁰ and k_v conditions was 6.64 g/l (Y_p/s: 0.27), which was about 1.5 times higher than the maximum concentration obtained in the aerated-flask.     

Low spring constant regulates P-selectin-PSGL-1 bond rupture

Forced dissociation of selectin-ligand bonds is crucial to such biological processes as leukocyte recruitment, thrombosis formation, and tumor metastasis. Although the bond rupture has been well known at high loading rate r_f (≥10² pN/s), defined as the product of spring constant k and retract velocity v, how the low r_f (<10² pN/s) or the low k regulates the bond dissociation remains unclear. Here an optical trap assay was used to quantify the bond rupture at r_f ≤ 20 pN/s with low k (∼10⁻³-10⁻² pN/nm) when P-selectin and P-selectin glycoprotein ligand 1 (PSGL-1) were respectively coupled onto two glass microbeads. Our data indicated that the bond rupture force f retained the similar values when r_f increased up to 20 pN/s. It was also found that f varied with different combinations of k and v even at the same r_f. The most probable force, f*, was enhanced with the spring constant when k < 47.0 × 10⁻³ pN/nm, indicating that the bond dissociation at low r_f was spring constant dependent and that bond rupture force depended on both the loading rate and the mechanical compliance of force transducer. These results provide new insights into understanding the P-selectin glycoprotein ligand 1 bond dissociation at low r_f or k.     

Light-induced slow changes in chlorophyll a fluorescence in isolated chloroplasts: Effects of magnesium and phenazine methosulfate

We have investigated the possible relationships between the cation-induced and phenazine methosulfate (PMS)-induced fluorescence changes and their relation to light induced conformational changes of the thylakoid membrane.1. In isolated chloroplasts, PMS markedly lowers the quantum yield of chlorophyll a fluorescence (φ_f) when added either in the presence or the absence of dichloro-phenyldimethylurea (DCMU). In contrast, Mg²⁺ causes an increase in φ_f. However, these effects are absent in isolated chloroplasts fixed with glutaraldehyde that retain (to a large extent) the ability to pump protons, suggesting that structural alteration of the membrane—not the pH changes—is required for the observed changes in φ_f. The PMS triggered decrease in φ_f is not accompanied by any changes in the emission (spectral) characteristics of the two pigment systems, whereas room temperature emission spectra with Mg²⁺ and Ca²⁺ show that there is a relative increase of System II to System I fluorescence.2. Washing isolated chloroplasts with 0.75 mM EDTA eliminates (to a large extent) the PMS-induced quenching and Mg²⁺-induced increase of φ_f, and these effects are not recovered by the further addition of dicyclohexyl carbodiimide. It is known that washing with EDTA removes the coupling factor, and thus, it seems that the coupling factor is (indirectly) involved in conformational change of thylakoid membranes leading to fluorescence yield changes.3. In purified pigment System II particles, neither PMS nor Mg²⁺ causes any change in φ_f. Our data, taken together with those of the others, suggest that a structural modification of the thylakoid membranes (not macroscopic volume changes of the chloroplasts) containing both Photosystems I and II is necessary for the PMS-induced quenching and Mg²⁺-induced increase of φ_f. These two effects can be explained with the assumption that the PMS effect is due to an increase in the rate of internal conversion (k_h), whereas the Mg²⁺ effect is due to a decrease in the rate of energy transfer (k_t), between the two photosystems.4. From the relative ratio of φ_f with DCMU and DCMU plus Mg²⁺, we have calculated k_t (the rate constant of energy transfer between Photosystems II and I to be 4.2·10⁸ s^?1, and φ_t (quantum yield of this transfer) to be 0.12.