Évaluation de l’exposition des travailleurs du service de radioimmunoanalyse du Bénin aux rayonnements ionisants de l’iode 125

Occupational radiation dose of staff handling ¹²⁵I assessment at the Benin radioimmunoassay laboratory, have been undertaken from October 2012 to April 2013 to determine level of radiation safety. Equivalent dose to skin, whole body and extremities, were measured by the mean of thermoluminescence dosimetry. Firstly, three permanent workers and two students were provided with finger ring dosimeters to wear at index finger base of both hands. Ring dosimeters were used for four months. Secondly, three permanent workers and three students were provided with badge dosimeters to wear at the chest level. Badge dosimeters were renewed monthly for six months. The exposed ring and badge dosimeters were evaluated in Ghana. Permanent workers highest average equivalent dose received at index finger base of both hands was 142.75 ± 89.54, microSV/2 months and that of students was 34.69 ± 29.23, microSV/2 months. Workers skin exposure was below one third of prescribed dose limits for permanents workers (500mSv/yr) and students (150mSv/yr). Whole body exposure, expressed in mSv/month, of permanent workers and students, respectively ranged from 0.12 to 0.23 and from 0.11 to 0.16. Radio-immuno-assay laboratory workers are weakly exposed to ionizing radiation. They are safe from deterministic effect risk.     

The effect of x-radiation upon the quality and fertility of stallion semen

The exposure that stallion semen might receive during examination using an airport x-ray security screening system was found to be between 0.5 and 1.0 micro Sieverts (μSv). Ejaculates from 2 stallions were diluted 1:4 (volume:volume) using a nonfat dried milk-glucose extender. A total of 6 ejaculates from each stallion was collected, and each ejaculate was divided into 3 aliquots and these were then exposed to x-radiation at a dose of 0, 1.0, or 10.0 (μSv. Semen quamy was examined immediately post exposure, and the aliquots were then placed into a water bath at 37 °C, after which sample longevity was evaluated.In a second trial, 3 groups of 8 pony mares were inseminated with semen that had been exposed to x-radiation at doses of 0, 1.0, or 10.0 μSv. An entire ejaculate was irradiated and inseminated into each mare on one occasion during estrus, based upon ultrasonographic evaluation of the reproductive tract.After exposure to x-radiation there were no differences among the 3 treatment groups for spermatozoal motility, morphology, or longevity. The 14-d pregnancy rates for the 3 treatment groups were 0 μSv (7 mares), 1.0 μSv (8 mares), and 10.0 μSv (7 mares). One mare (0 (μSv) aborted at 65 d of pregnancy; 21 mares had a pregnancy of normal length, with each delivering a foal at term, although 1 foal died at parturition (1.0 μSv).These findings indicate that the exposure of stallion spermatozoa to x-radiation up to doses of 10 μSv does not have deleterious effects upon spermatozoal motility, morphology, longevity or fertility. The exposure received during examination using an x-ray security screening system is likely to be lower than this dose.     

Intestinal microbiota as novel biomarkers of prior radiation exposure

There is an urgent need for rapid, accurate, and sensitive diagnostic platforms to confirm exposure to radiation and estimate the dose absorbed by individuals subjected to acts of radiological terrorism, nuclear power plant accidents, or nuclear warfare. Clinical symptoms and physical dosimeters, even when available, do not provide adequate diagnostic information to triage and treat life-threatening radiation injuries. We hypothesized that intestinal microbiota act as novel biomarkers of prior radiation exposure. Adult male Wistar rats (n = 5/group) received single or multiple fraction total-body irradiation of 10.0 Gy and 18.0 Gy, respectively. Fresh fecal pellets were obtained from each rat prior to (day 0) and at days 4, 11, and 21 post-irradiation. Fecal microbiota composition was determined using microarray and quantitative PCR (polymerase chain reaction) analyses. The radiation exposure biomarkers consisted of increased 16S rRNA levels of 12 members of the Bacteroidales, Lactobacillaceae, and Streptococcaceae after radiation exposure, unchanged levels of 98 Clostridiaceae and Peptostreptococcaceae, and decreased levels of 47 separate Clostridiaceae members; these biomarkers are present in human and rat feces. As a result of the ubiquity of these biomarkers, this biomarker technique is non-invasive; microbiota provide a sustained level of reporting signals that are increased several-fold following exposure to radiation, and intestinal microbiota that are unaffected by radiation serve as internal controls. We conclude that intestinal microbiota serve as novel biomarkers of prior radiation exposure, and may be able to complement conventional chromosome aberrational analysis to significantly enhance biological dose assessments.     

Comparison of EPR occupational lifetime external dose assessments for Mayak nuclear workers and film badge dose data

The Mayak worker cohort is one of the major sources of information on health risks due to protracted exposures to plutonium and external ionizing radiation. Electron paramagnetic resonance (EPR) measurements in tooth enamel in combination with personal dose monitoring can help to improve external dose assessment for this cohort. Here, the occupational lifetime external exposure was evaluated individually for 44 nuclear workers of three plants of the Mayak Production Association by EPR measurements of absorbed doses in collected tooth enamel samples. Analysis included consideration of individual background doses in enamel and dose conversion coefficients specific for photon spectra at selected work areas. As a control, background doses were assessed for various age groups by EPR measurements on teeth from non-occupationally exposed Ozyorsk residents. Differences in occupational lifetime doses estimated from the film badges and from enamel for the Mayak workers were found to depend on the type of film badge and the selected plant. For those who worked at the radiochemical processing plant and who were monitored with IFK film badges, the dose was on average 570 mGy larger than estimated from the EPR measurements. However, the average difference was found to be only −4 and 6 mGy for those who were monitored with IFKU film badges and worked at the reactor and the isotope production plant respectively. The discrepancies observed in the dose estimates are attributed to a bias in film badge evaluation.N. El-Faramawy: On leave from Department of Physics, Faculty of Science, Ain Shams University, 65511 Abbassia, Cairo, Egypt.     

Use of chemical dosimetry for comparison of ultrasound and ionizing radiation effects on cavitation

A comparison of the effects of ultrasound produced by low- and high-frequency ultrasonic apparatuses upon biological systems is one of the basic problems when studying ultrasound cavitation effects. One possibility for how to compare these effects is the indirect method which uses well-known physical quantities characterizing the interaction of ionizing radiation with matter and which also converts these quantities to one common physical quantity. The comparison was performed with two methods applied to the chemical dosimetry of ionizing radiation. The first method employed a two-component dosimeter which is composed of 50 % chloroform and 50 % re-distilled water (i.e. Taplin dosimeter). The other method used a modified iodide dosimeter prepared from a 0.5 M potassium iodide solution. After irradiation or ultrasound exposure, measurable chemical changes occurred in both dosimeters. The longer the exposure, the greater the chemical changes. These effects are described by the relationship of these changes versus the exposure times in both dosimeters. The UZD 21 ultrasonic disintegrator (with a frequency of 20 kHz, 50 % power output) was used as a low-frequency ultrasound source, and the BTL-07 therapeutic instrument (with a frequency of 1 MHz and intensity of 2 W/cm2) was used as a high-frequency cavitation ultrasound source. For comparison, a 60 Co gamma source was applied (60 Co, gamma energies of 1.17 and 1.33 MeV, activity of 14 PBq). Results of this study have demonstrated that the sonochemical products are generated during exposure in the exposed samples of both dosimeters for all apparatuses used. The amount of these products depends linearly upon the exposure time. The resulting cavitation effects were recalculated to a gray-equivalent dose (the proposed unit is cavitation gray [cavitGy]) based on the sonochemical effects compared to the effects of ionizing radiation from the 60 Co source.     

Reasons for requesting radiographs in an accident department.

A prospective study of 500 patients was performed to determine the reasons for requesting radiographs in an accident and emergency department. Most examinations were requested either to confirm a clinically suspected abnormality or because of difficulty in excluding a significant bone injury on clinical grounds alone. Several requests were also made to reassure the patient. Medicolegal reasons were relatively few, and those made purely because the doctor feared litigation probably accounted for only 5% of requests. Undue emphasis on the medicolegal aspects of accident and emergency radiography in the United Kingdom is unhelpful in that it directs attention away from the real reasons for x-ray referral. Although a reduction in the number of x-ray examinations is desirable on the grounds of expense and radiation exposure it is likely to be obtained only by improving experience and acumen in the clinical assessment of injuries.     

The Role of the Pineal Body in Ectotherm Thermoregulation

The pineal complex may be a part of the sensory and centralneural system controlling thermoregulatory behavior. The pinealand parapineal organs of some ectotherms appear to functionas radiation dosimeters, regulating exposure to sunlight. Physiologicalthermoregulation may be influenced by the pineal complex throughcardiovascular adjustments or metabolic rates. Additionally,the pineal organ may exert thermoregulatory effects throughthe control of brain electrolytes. While the precise mechanismsof action remain to be defined, it is clear that pineal-parapinealorgans participate in thermoregulatory adjustments by actingupon the central nervous system.     

Radiosensitivity of the helper cell function.

The helper function of T cells primed and irradiated in vivo was tested in vitro by the Mishell-Dutton technique. Spleen cells from mice carrier-primed with HRBC and exposed to 50 to 2000 rads of x-radiation were assayed for their ability to help syngeneic normal spleen cells to mount an in vitro anti-hapten antibody response after stimulation with the conjugate TNP-HRBC. The anti-TNP response was evaluated by the Jerne technique. The helper activity was titrated by adding graded numbers of carrier-primed spleen cells to a constant number of normal spleen cells. The slope of the initial linear portion of the response-cell dose titration curve was taken as an estimated of the helper activity and found to decrease with increasing the x-ray dose. The curve describing the remaining helper activity as a function of the radiation dose shows the presence of two components, one radiosensitive, the other, radioresistant. This suggests the existence either of helper cells at different stages of activation or of two cell subpopulations participating in the helper function.     

THE EFFECT OF CORTISONE ON DNA CONTENT OF RAT HEPATOCYTES

Rats were treated with cortisone, x-radiation, and both agents in combination, and the effect noted on the DNA content of hepatocytes. Nuclei were enumerated both in whole liver homogenates and following isolation. The incorporation of P³² into DNA was also studied in relation to these agents. The following observations were made:— 1.The DNA content of nuclei fell both during cortisone administration and following x-radiation. In the former instance, the fall was progressive with continuing administration of hormone; in the latter instance, there was a return to normal 5 days after radiation. 2. Cortisone administration to x-radiated rats caused a fall in DNA/nucleus and prevented the return to normal at 5 days. 3. There was no evidence that the effects of cortisone and x-rays were additive in reducing DNA/nucleus. 4. These data indicate an alteration in DNA/nucleus, but simple changes in ploidy cannot be excluded. Either explanation requires that the agents used affect the DNA of non-regenerating nuclei. 5. Cortisone interfered with the incorporation of P³² into the DNA of regenerating liver. Only a small effect on DNA synthesis in resting liver was observed with cortisone or x-radiation. 6. DNA content of nuclei returned to normal 5 days after x-radiation and 3 days after discontinuance of cortisone. Slight increase in the incorporation of P³² by DNA was observed during recovery phases. 7. The hypothesis is proposed that the apparent losses and increases in DNA/nucleus were due to depolymerization and repolymerization of DNA. Following x-radiation and/or cortisone administration, it is proposed that some DNA is depolymerized and becomes cold acid-soluble and dissociated from organized chromatin. Later, conditions are such that this degraded DNA is repolymerized. 8. These data might be interpreted to indicate that a portion of the DNA is not essential to cell integrity; alternatively, there may be two or more species of DNA, one of which is more readily affected by the agents investigated in the present report.     

Calibration of x-ray densitometry for the measurement of coral skeletal density

A new method is proposed for the measurement of coral skeletal density by x-radiography. X-radiographs were made of sections cut from skeletons of massive corals of the genus Porites. Included on the x-ray film with each specimen were an aluminium step-wedge, a set of aragonite standards and several aluminium bars, all of measured thickness and density. The images on the developed x-ray film were scanned with a microdensitometer. Semilogarithmic plots of microdensitometer output voltage vs. thickness of the aluminium and aragonite standards provided characteristic curves, with initial linear slopes which were defined as relative linear absorption coefficients. These coefficients varied with the x-ray exposure and microdensitometer measurement conditions; however, they were consistent within one set of conditions. The relative linear absorption coefficients and densities of the standards, together with data for thickness of standard vs. film exposure, can be used to determine the density of coral specimens at points along microdensitometer traverses of their x-ray images. The bars of aluminium can be used to correct for the non-uniform irradiation of the x-ray film which is characteristic of x-ray machines.Contribution No. 299 from the Australian Institute of Marine Science     

In Situ μGISAXS: II. Thaumatin Crystal Growth Kinetic

The formation of thaumatin crystals by Langmuir-Blodgett (LB) film nanotemplates was studied by the hanging-drop technique in a flow-through cell by synchrotron radiation micrograzing-incidence small-angle x-ray scattering. The kinetics of crystallization was measured directly on the interface of the LB film crystallization nanotemplate. The evolution of the micrograzing-incidence small-angle x-ray scattering patterns suggests that the increase in intensity in the Yoneda region is due to protein incorporation into the LB film. The intensity variation suggests several steps, which were modeled by system dynamics based on first-order differential equations. The kinetic data can be described by two processes that take place on the LB film, a first, fast, process, attributed to the crystal growth and its detachment from the LB film, and a second, slower process, attributed to an unordered association and conversion of protein on the LB film.     

Radiation protection of operators and patients in a hybrid Angio-MR suite

This work investigates the patient eye lens dose and x-ray scatter to the operator expected for a proposed hybrid Angio-MR concept. Two geometries were simulated for comparative assessment: a standard C-arm device for neuro-angiography applications and an innovative hybrid Angio-MR system concept, proposed by Siemens Healthineers. The latter concept is based on an over-couch x-ray tube and a detector inside an MRI system, with the aim of allowing combined, simultaneous MRI and x-ray imaging for procedures such as neurovascular interventions (including x-ray fluoroscopy and angiography imaging, 3D imaging, diffusion, and perfusion). To calculate the scattered radiation dose to the physician, Monte Carlo simulations were performed. Dose estimates of simplified models of the brain and eyes of both the patient and the physician and of the physician’s torso and legs have been calculated. A number of parameters were varied in the simulation including x-ray spectrum, field of view (FOV), x-ray tube angulation, presence of shielding material and position of the physician. Additionally, 3D dose distributions were calculated in the vertical and horizontal planes in both setups. The patient eye lens dose was also calculated using a detailed voxel phantom and measured by means of thermoluminescent dosimeters (TLDs) to obtain a more accurate estimate. Assuming the same number of x-rays and the same size of the irradiated area on the patient’s head, the results show a significant decrease in the scattered radiation to the physician for the Angio-MR system, while large increases, depending on setup, are expected to patient eye lens dose.     

Radiation Hazards Control in Survival Operations in the Event of a Nuclear War

The concepts of radiation protection in survival operations are explained, and procedures devised to control radiation hazards for the protection of the population and maintenance of the operating efficiency of survival operations personnel are presented.Radiation protection is a command function. The medical responsibility is to provide advice on the probable effects of radiation exposure in the light of existing knowledge of these effects in man.The major hazard is that of external exposure to penetrating gamma radiation. Radiation exposure guides indicate that persons may be exposed to not more than 100 r whole body radiation in a six-week period, or 200 r whole body radiation in a period in excess of six weeks, without loss of operational efficiency. Beta radiation from fallout deposited on skin or clothing may produce burns, but these injuries will not be incapacitating and can be controlled by simple procedures.The internal hazard is mainly from ingestion of food or water contaminated with radioactive material. For protection, only canned or packaged foods and water from covered or deep wells are consumed during the early days after a nuclear attack.     

Protein-facilitated gold nanoparticle formation as indicators of ionizing radiation

The use of X-ray radiation in radiotherapy is a common treatment for many cancers. Despite several scientific advances, determination of radiation delivered to the patient remains a challenge due to the inherent limitations of existing dosimeters including fabrication and operation. Here, we describe a colorimetric nanosensor that exhibits unique changes in color as a function of therapeutically relevant radiation dose (3–15 Gy). The nanosensor is formulated using a gold salt and maltose-binding protein as a templating agent, which upon exposure to ionizing radiation is converted to gold nanoparticles. The formation of gold nanoparticles from colorless precursor salts renders a change in color that can be observed visually. The dose-dependent multicolored response was quantified through a simple ultraviolet–visible spectrophotometer and the peak shift associated with the different colored dispersions was used as a quantitative indicator of therapeutically relevant radiation doses. The ease of fabrication, visual color changes upon exposure to ionizing radiation, and quantitative read-out demonstrates the potential of protein-facilitated biomineralization approaches to promote the development of next-generation detectors for ionizing radiation.     

Fas is involved in the p53-dependent apoptotic response to ionizing radiation in mouse testis

Apoptosis induced in male germ cells following ionizing radiation is dependent on functional p53 (Trp53) being present. We sought to determine whether Fas (Tnfrsf6/CD95/APO-1), an apoptotic factor, is involved in this p53-dependent germ cell death. In p53 knock-out mice exposed to 5 Gy of x-radiation, germ cells were protected from cell death, as assessed by counting apoptotic seminiferous tubules 12 h following radiation. Similarly, spermatid head counts in p53 knock-out mice remained near normal 29 days after exposure to 0.5 Gy of radiation, whereas wild-type animals had a more than twofold reduction in spermatid head counts. Fas mRNA expression remained at pretreatment levels in p53 knock-out mice; however, Fas increased in a time-dependent manner in wild-type mice following exposure to 5 Gy of radiation, indicating that radiation-induced Fas expression is p53-dependent. The functional significance of Fas involvement was demonstrated when lpr(cg) mice, having a nonfunctional Fas receptor, were exposed to 5 Gy of radiation; the number of apoptotic seminiferous tubules 12 h following radiation was significantly reduced compared to that of wild-type mice. Additionally, lpr(cg) mice exposed to 0.5 Gy of radiation had increased spermatid head counts 29 days following radiation compared to wild-type mice. Interestingly, gld mice with a non-functional Fas ligand (Tnfsf6/FasL/CD95L) were as sensitive to radiation as wild-type animals, and levels of FasL mRNA were not affected by radiation treatment. These results indicate that apoptosis and up-regulation of Fas following radiation are both p53-dependent events. Although Fas is necessary, in part, for radiation-induced p53-dependent apoptosis, FasL is not.     

Mesure de l’exposition externe du personnel du service de médecine nucléaire de l’hôpital Ibn Sina de Rabat au cours de la manipulation des radiopharmaceutiques

The nuclear medicine department of Ibn Sina hospital was created in 1965. Its first location was in radiology department, and then it was transferred in 1977 to the old emergency unit. The conception was not originally dedicated for a nuclear medicine department. To reassure the medical staff about their exposition during radiopharmaceutical manipulation, a dosimetric study was performed in collaboration with the National center of radiation protection (CNRP). Dosimetric monitoring was conducted for 3 months with the use of passive thermoluminescent dosimeters, (Harshow, 6600 plus), covering all categories of staff. The administration of 153 Samarium has been also studied. Other cases of people concerned our study: a pregnant woman doctor, whose exposure of the unborn child must be reduced as much as possible, and a woman from a private company, working occasionally in the department. To control the conditions imposed on all activities requiring exposure to ionizing radiation, we evaluated the dose at the extremities of operators with the use of ring dosimeters (Harshow 6600 plus) and the dose on the ambient environment of staff. This experiment has shown exposure levels below legal limits, without been negligible for some workstations. Technicians equivalent doses results justify the wearing of dosimeter rings as a complementary dosimeter in nuclear medicine department. Finally, monitoring ambient dosimetry showed that the environment measurements are in keeping with national and international norms. The Medical staff was very interested by this study, which demonstrates that radiation protection standards are respected through awareness, basic training, continuing education and information. The first author is particularly involved in the practical procedures in order to respect radiation protection recommendations for restraining staff annual chest and extremities dosimetry figures within regulations values.     

Radiation dose of digital radiography (DR) versus micro-dose x-ray (EOS) on patients with adolescent idiopathic scoliosis: 2016 SOSORT- IRSSD “John Sevastic Award” Winner in Imaging Research

Background

Patients with adolescent idiopathic scoliosis (AIS) frequently receive x-ray imaging at diagnosis and subsequent follow monitoring. The ionizing radiation exposure has accumulated through their development stage and the effect of radiation to this young vulnerable group of patients is uncertain. To achieve the ALARA (as low as reasonably achievable) concept of radiation dose in medical imaging, a slot-scanning x-ray technique by the EOS system has been adopted and the radiation dose using micro-dose protocol was compared with the standard digital radiography on patients with AIS.

Methods

Ninety-nine participants with AIS underwent micro-dose EOS and 33 underwent standard digital radiography (DR) for imaging of the whole spine. Entrance-skin dose was measured using thermoluminescent dosimeters (TLD) at three regions (i.e. dorsal sites at the level of sternal notch, nipple line, symphysis pubis). Effective dose and organ dose were calculated by simulation using PCXMC 2.0. Data from two x-ray systems were compared using independent-samples t-test and significance level at 0.05. All TLD measurements were conducted on PA projection only. Image quality was also assessed by two raters using Cobb angle measurement and a set of imaging parameters for optimization purposes.

Results

Entrance-skin dose from micro-dose EOS system was 5.9–27.0 times lower at various regions compared with standard DR. The calculated effective dose was 2.6?±?0.5 (μSv) and 67.5?±?23.3 (μSv) from micro-dose and standard DR, respectively. The reduction in the micro-dose was approximately 26 times. Organ doses at thyroid, lung and gonad regions were significantly lower in micro-dose (p?<?0.001). Data were further compared within the different gender groups. Females received significantly higher (p?<?0.001) organ dose at ovaries compared to the testes in males. Patients with AIS received approximately 16–34 times lesser organ dose from micro-dose x-ray as compared with the standard DR. There was no significant difference in overall rating of imaging quality between EOS and DR. Micro-dose protocol provided enough quality to perform consistent measurement on Cobb angle.

Conclusions

Entrance-skin dose, effective dose and organ dose were significantly reduced in micro-dose x-ray. The effective dose of a single micro-dose x-ray (2.6 μSv) was less than a day of background radiation. As AIS patients require periodic x-ray follow up for surveillance of curve progression, clinical use of micro-dose x-ray system is beneficial for these young patients to reduce the intake of ionizing radiation.

     

Radiation dose measurements in a dental orthopantogram unit using indigenously developed optically stimulated luminescence dosimeters

Dental orthopantogram (OPG)/cone beam computed tomography (CBCT) scanners are gaining popularity due to their 3D imaging with multiplanar view that provides clinical benefits over conventional dental radiography systems. Dental OPG/CBCT provides optimal visualization of adjacent overlaying anatomical structures that will be superpositioned in any single projection. The characteristics of indigenously developed optically stimulated luminescence dosimeters, namely, aluminium oxide doped with carbon (Al₂O₃:C), lithium magnesium phosphate doped with terbium and boron (LiMgPO₄:Tb,B) and lithium calcium aluminium fluoride doped with europium and yttrium (LiCaAlF₆:Eu,Y) were evaluated for their use in dental dosimetry. The dose?response of these dosimeters was studied at X‐ray energies 60 kV, 70 kV and 81 kV. Radiation doses were also measured using Gafchromic film for comparison. Radiation dose was measured at eight different locations of a polymethyl methacrylate (PMMA) head phantom including eyes. The optically stimulated luminescence (OSL) sensitivity of LiMgPO₄:Tb,B is about 1.5 times and LiCaAlF₆:Eu, is about 20 times higher than the sensitivity of Al₂O₃:C. It was found that measured radiation doses by the three optically stimulated luminescence dosimeters (OSLDs) and Gafchromic film in the occipital region (back side) of a PMMA phantom, were consistent but variations in dose at other locations were significantly higher. The three OSLDs used in this study were found to be suitable for radiation dose measurement in dental units.     

Natural and sexual selection on a plumage signal of status and on morphology in house sparrows,Passer domesticus

Temporal patterns of natural and sexual selection on male badge size and body traits were studied in a population of house sparrows, Passer domesticus. Badge size was a heritable trait as revealed by a significant father-son regression. Survival during autumn dispersal and winter was not related to badge size or body traits in yearling male house sparrows. Badges that signal dominance status were affected positively by directional selection for mating. Adult male house sparrows suffered an opposing selection pressure on badge size during autumn. Contrary to males, female house sparrows did not experience significant directional or stabilizing selection on any body trait. Directional sexual selection on male badge size due to female choice moves male sparrows away from their survival optimum. Opposing directional natural selection on badge size due to autumn mortality caused by predation maintains a stable badge size.     

Characterization of conditions required for X-Ray diffraction experiments with protein microcrystals

The x-ray exposure at which significant radiation damage occurs has been quantified for frozen crystals of bacteriorhodopsin. The maximum exposure to approximately 11-keV x-rays that can be tolerated for high-resolution diffraction experiments is found to be approximately 10(10) photons/microm(2), very close to the value predicted from limits that were measured earlier for electron diffraction exposures. Sample heating, which would further reduce the x-ray exposure that could be tolerated, is not expected to be significant unless the x-ray flux density is well above 10(9) photons/s-microm(2). Crystals of bacteriorhodopsin that contain approximately 10(11) unit cells are found to be large enough to give approximately 100 high-resolution diffraction patterns, each covering one degree of rotation. These measurements are used to develop simple rules of thumb for the minimum crystal size that can be used to record x-ray diffraction data from protein microcrystals. For work with very small microcrystals to be realized in practice, however, it is desirable that there be a significant reduction in the level of background scattering. Background reduction can readily be achieved by improved microcollimation of the x-ray beam, and additional gains can be realized by the use of helium rather than nitrogen in the cold gas stream that is used to keep the protein crystals frozen.