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PubMed1994

The circumstellar environment of IRAS 05338-0624.

Millimeter continuum and spectral line observations with 10", 30", and 60" resolution are used to characterize the structure and chemistry of the gas around the young, embedded star, IRAS 05338-0624. On arcminute scales, emission from dense gas tracers outline an isolated condensation centered on the IRAS source position. The condensation is characterized by a size of approximately 60", a density of 2 x 10(5) cm-3, and a virial mass of 40 M solar. Interferometric CS J = 2-1 observations show two peaks, one toward the continuum peak and the other toward a position 14" west and 8" south. Single-dish maps of SO, CH3OH, and SiO show pronounced wing emission to the west of the IRAS source, which interferometer observations reveal to be a compact region of outflow activity. CS emission as redshifted and blueshifted velocities reveals a bipolar outflow oriented with a position angle of 45 degrees, while SiO emission appears to be tracing a fast shock interaction region at the CS red-lobe peak, 14" west and 8" south of the IRAS source. Finally, H13CO+ emission traces clumps of quiescent gas toward the IRAS source and adjacent to the blue lobe of the outflow. Column densities and molecular fractional abundances are derived to explore the interaction between the surrounding condensation and the young stellar object. We find evidence for gas phase depletions within the overall condensation in several gas tracers (CO, CS, HCN, SO) but not in the region immediately around the young stellar object. Enhanced abundances of SO, CH3OH, and SiO (by factors of 4, >100, >1000, respectively) are observed in the shocked gas; these enhancements may be explained in terms of a nondissociative shock liberating mantle materials that contain some amount of refractory materials, a moderate velocity dissociative shock in which only minor sputtering of Si occurs, or a shock that impacts surrounding material with a range of speeds.

Astronomy↗
PubMed1993

Aqueous geochemistry on early Mars.

A geochemical cycle model is presented for the interaction between the atmosphere, hydrosphere, and regolith of Mars. It was developed to study how this interaction might have produced the present Martian environment from a primitive Martian environment much like that of the primitive Earth. The model is a simple system, consisting of an unweathered starting material (calcium-bearing and magnesium-bearing silicates), a CO2 atmosphere, an ocean of water in contact with both the atmosphere and the unweathering starting material, and both calcite and dolomite precipitates. Several interesting points arise from this model. A 1-bar CO2 atmosphere can be removed by carbonate precipitation alone in about half a billion years. This is roughly fifty times longer than earlier estimates, which were not based on time-varying models (Fanale et al., 1982; Carr, 1986; Pollack et al., 1987). One of the chief problems in Martian geology has been how to explain the large number and wide variety of surface features that were apparently formed by aqueous erosion. This longer atmospheric lifetime may be enough to explain the large number of channels seen on older Martian terrain. If the atmosphere started out with more than 1 bar of CO2, it would take correspondingly longer to remove it. If there should be no other means to remove CO2 from the atmosphere, this long time constant would indicate that the atmosphere could never have contained more than a few bars of CO2, or else there would still be remnants present today. The increase in alkalinity of the ocean as the atmosphere disappears, even without the effects of reduction in the amount of water available, indicates that evaporite deposits may have formed on Mars. If these deposits are still present, they may even yet contain some liquid water.

Atmosphere↗
PubMed1999

Growing crops for space explorers on the moon, Mars, or in space.

An option in the long-duration exploration of space, whether on the Moon or Mars or in a spacecraft on its way to Mars or the asteroids, is to utilize a bioregenerative life-support system in addition to the physicochemical systems that will always be necessary. Green plants can use the energy of light to remove carbon dioxide from the atmosphere and add oxygen to it while at the same time synthesizing food for the space travelers. The water that crop plants transpire can be condensed in pure form, contributing to the water purification system. An added bonus is that green plants provide a familiar environment for humans far from their home planet. The down side is that such a bioregenerative life-support system--called a controlled environment life-support system (CELSS) in this paper--must be highly complex and relatively massive to maintain a proper composition of the atmosphere while also providing food. Thus, launch costs will be high. Except for resupply and removal of nonrecycleable substances, such a system is nearly closed with respect to matter but open with respect to energy. Although a CELSS facility is small compared to the Earth's biosphere, it must be large enough to feed humans and provide a suitable atmosphere for them. A functioning CELSS can only be created with the help of today's advanced technology, especially computerized controls. Needed are energy for light, possibly from a nuclear power plant, and equipment to provide a suitable environment for plant growth, including a way to supply plants with the necessary mineral nutrients. All this constitutes the biomass production unit. There must also be food preparation facilities and a means to recycle or dispose of waste materials and there must be control equipment to keep the facility running. Humans are part of the system as well as plants and possibly animals. Human brain power will often be needed to keep the system functional in spite of the best computer-driven controls. The particulars of a CELSS facility depend strongly on where it is to be located. The presence of gravity on the Moon and Mars simplifies the design for a facility on those bodies, but a spacecraft in microgravity is a much more challenging environment. One problem is that plants, which are very sensitive to gravity, might not grow and produce food in the virtual absence of gravity. However, the experience with growing super-dwarf wheat in the Russian space station Mir, while not entirely successful because of the sterile wheat heads, was highly encouraging. The plants grew well for 123 days, producing more biomass than had been produced in space before. This was due to the high photon flux available to the plants and the careful control of substrate moisture. The sterile heads were probably due to the failure to remove the gaseous plant hormone, ethylene, from the Mir atmosphere. Since ethylene can easily be removed, it should be possible to grow wheat and other crops in microgravity with the production of viable seeds. On the ground Biosphere-2 taught us several lessons about the design and construction of a CELSS facility, but Bios-3 came much closer to achieving the goals of such a facility. Although stability was never completely reached, Bios-3 was much more stable than Biosphere-2 apparently because every effort was made to keep the system simple and to use the best technology available to maintain control. Wastes were not recycled in Bios-3 except for urine, and inedible plant materials were incinerated to restore CO2 to the atmosphere. Since much meat (about 20% of calories) was imported, closure in the Bios-3 experiments was well below 100%. But then, a practical CELSS on the Moon might also depend on regular resupply from Earth. Several important lessons have been learned from the CELSS research described in this review.

Agriculture↗
PubMed2002

Radiations in space: risk estimates.

The complexity of radiation environments in space makes estimation of risks more difficult than for the protection of terrestrial populations. In deep space the duration of the mission, position in the solar cycle, number and size of solar particle events (SPE) and the spacecraft shielding are the major determinants of risk. In low-earth orbit missions there are the added factors of altitude and orbital inclination. Different radiation qualities such as protons and heavy ions and secondary radiations inside the spacecraft such as neutrons of various energies, have to be considered. Radiation dose rates in space are low except for short periods during very large SPEs. Risk estimation for space activities is based on the human experience of exposure to gamma rays and to a lesser extent X rays. The doses of protons, heavy ions and neutrons are adjusted to take into account the relative biological effectiveness (RBE) of the different radiation types and thus derive equivalent doses. RBE values and factors to adjust for the effect of dose rate have to be obtained from experimental data. The influence of age and gender on the cancer risk is estimated from the data from atomic bomb survivors. Because of the large number of variables the uncertainities in the probability of the effects are large. Information needed to improve the risk estimates includes: (1) risk of cancer induction by protons, heavy ions and neutrons: (2) influence of dose rate and protraction, particularly on potential tissue effects such as reduced fertility and cataracts: and (3) possible effects of heavy ions on the central nervous system. Risk cannot be eliminated and thus there must be a consensus on what level of risk is acceptable.

Aerospace Medicine↗
PubMed1965

Procedures necessary for the prevention of planetary contamination.

Studies of microbial survival in simulated deep-space conditions have established that these conditions will not sterilize contaminated spacecraft. Likewise, data presented at this and previous meetings of COSPAR have shown that many terrestrial microbes, particularly anaerobic sporeformers, readily tolerate simulated Martian environments. Any spacecraft landing on Mars must have a low probability of harboring any terrestrial organisms, especially sporeformers, if the goals of exobiology are not to be compromised. Before the planning, production and monitoring of sterile spacecraft can be undertaken, the theory and principles of sterilization and their application to the special problem of spacecraft sterilization must be understood. The response of a given population of microorganisms to sterilizing agents is affected by several factors; these factors in turn regulate the kinetics of the killing process. The only processes that achieve both surface and interior sterilization are heat and radiation. Of these, dry heat is the agent of choice. If all parts of the spacecraft are assembled under the most rigorous conditions of cleanliness, the capsule can be brought to terminal sterilization containing a total of not more than 10(5) viable organisms. These can be killed by exposure to 135 degrees for 24 hours with a confidence of greater than 10(4). Spacecraft must be specifically designed to withstand such heat sterilization without reduction of reliability. The number of labile parts must be kept to a minimum, sterilized by methods other than heat, and inserted into the sterilized spacecraft by sterile techniques. The entire process must be under very tight control with tests and records of every detail. The sterilized spacecraft must be encapsulated and remain therein during final testing and launch until the capsule is opened in deep space. The design and production engineer must understand how all these procedures will affect the spacecraft, and the biologist must constantly monitor and educate the engineers on the scope and limitations of these various sterilizing procedures.

Colony Count, Microbial↗
PubMed1996

Absorbed dose measurements on LDEF and comparisons with predictions.

The radiation environment on LDEF was monitored by cumulative absorbed dose measurements made with TLDs at different locations and shielding depths. The TLDs were included in four experiments: A0015(a) Biostack, P0004 Seeds in Space and P0006 Linear Energy Transfer Spectrum Measurements at the trailing edge (west side) of the satellite; M0004 Fiber Optics Data Link at the leading edge (east side); and A0015(b) Biostack at the Earth side. The shielding depths varied between 0.48 and 15.4 g/cm2, Al equivalent. Both the directional dependence of trapped protons incident on the satellite and the shielding thickness were reflected in absorbed dose values. The trapped proton anisotropy was measured by TLDs at the east and west sides of LDEF. At the east side doses ranged from 2.10 to 2.58 Gy under shielding of 2.90 to 1.37 g/cm2 (M0004) while on the west side doses ranged from 2.66 to 6.48 Gy under shielding of 15.4 to 0.48 g/cm2 (P0006). The west side doses were more than a factor of two higher, where the vertical shielding thicknesses to space were equal. Other west side doses of 3.04 to 4.49 Gy under shielding of 11.7 to 3.85 g/cm2 (A0015(a)) and 2.91 to 6.64 Gy under shielding of 11.1 to 0.48 g/cm2 (P0004) generally agreed with the P0006 results. The Earth side doses of 2.41 to 3.93 Gy under shielding of 10.0 to 1.66 g cm2 (A0015(b)) were intermediate between the east side and west side doses. Calculations utilizing a model of trapped proton spectra were performed by Watts et al. (1993) and comparisons of dose measurement and calculations may be found in a companion paper (Armstrong et al., 1996).

Aluminum↗
PubMed1973

Survival of bacterial isolates exposed to simulated Jovian trapped radiation belt electrons and solar wind protons.

With missions to Jupiter, the spacecraft will be exposed for extended durations to solar wind radiation and the Jovian trapped radiation belt. This study is designed to determine the effect of these radiation environments on spacecraft bacterial isolates. The information can be used in the probability of contamination analysis for these missions. A bacterial subpopulation from Mariner Mars 1971 spacecraft (nine spore-forming and three non-spore-forming isolates) plus two comparative organisms, Staphylococcus epidermidis ATCC 17917 and a strain of Bacillus subtilis var. niger, were exposed to 2, 12, and 25 MeV electrons at different doses with simultaneous exposure to a vacuum of 1.3 x 10(-4) N m-2 at 20 and -20 degrees C. The radioresistance of the subpopulation was dependent on the isolate, dose and energy of electrons. Temperature affected the radioresistance of only the spore-forming isolates. Survival data indicated that spores were reduced approximately 1 log/1500 J kg-1 (10 J kg-1=1 krad), while non-spore-forming isolates (micrococci) were reduced 1.5-2 logs/1500 J kg-1 with the exception of an apparent radioresistant isolate whose resistance approached that of the spores. The subpopulation was found to be less resistant to lower energy than to higher energy electrons. The bacterial isolates were exposed to 3 keV protons under the same conditions as the electrons with a total fluence of 1.5 x 10(13) p cm-2 and a dose rate of 8.6 x 10(9) p cm-2 s-1. The results showed that only 20% of S. epidermidis and 45% of B. subtilis populations survived exposure to the 3 keV protons, while the mean survival of the spacecraft subpopulation was 45% with a range from 31.8% (non-spore-former) to 64.8% (non-spore-former). No significant difference existed between spore-forming and non-spore-forming isolates.

Bacillus subtilis↗
PubMed2003

Fundamental space radiobiology.

The unique feature of the space radiation environment is the dominance of high-energy charged particles (HZE or high LET radiation) emitted by the Sun and galactic sources, or trapped in the Van Allen radiation belts. These charged particles present a significant hazard to space flight crews, and accelerator-based experiments are underway to quantify the health risks due to unavoidable radiation exposure. There are three principal properties of charged particles that distinguish them from conventional radiation, i.e. gamma rays and x-rays. First, they have a defined range in matter rather than an exponential absorption profile. Second, they undergo nuclear reactions to produce secondary particles. Third, and most important, they deposit their energy along well-defined linear paths or tracks rather than diffuse fields. The structured energy deposition pattern interacts on multiple scales with the biological structures of DNA, cells and tissues to produce correlated patterns of damage that evade repair systems. Traditional concepts of dose and its associated normalization parameter, RBE (relative biological effectiveness), break down under experimental scrutiny, and probabilistic models of risk based on the number of particle traversals per cell may be more appropriate. Unique patterns of DNA damage, gene expression, mobilization of repair proteins, activation of cytokines and remodeling of cellular microenvironment are observed following exposure to high LET radiation. At low levels of exposure the communication of bioactive substances from irradiated to unirradiated "bystander" cells can amplify the damage and cause a significant deviation from linearity in dose vs. response relations. Under some circumstances, there is even a multigenerational delay in the expression of radiation-induced genetic damage (genomic instability) which is not strictly dose dependent. These issues and the experimental evidence derived from ground based experiments at particle accelerators are presented along with speculation about how modified inertial conditions might perturb homeostatic responses to radiation to further complicate risk assessment for space flight.

Animals↗
PubMed2003

Developmental biology of urodele amphibians in microgravity conditions.

Among the urodele amphibians, only Cynops pyrrhogaster and Pleurodeles waltl, two species of the Salamandridae family, were used in space experiments. The advantages for using urodeles reside (i) in reproduction: a few months after natural breeding, females can lay eggs in absence of males after a hormonal treatment, because spermatozoa were preserved in the cloacal pelvic glands of matted females, (ii) in the rate of development which is slower in Cynops and Pleurodeles than in the anuran Xenopus, (iii) in their physiological properties: they can live in a closed water container or in a moisturized environment, and they can fast during several days. Moreover, urodeles have an important phylogenetic interest. Many biological phenomena differ from those of anurans, such as fertilization events, the germ cell origin and the migration toward the differentiating gonads, and their regeneration capabilities. The main goals of the space experiments were to answer the following questions. On the one hand, does fertilization occur normally in microgravity? Is subsequent embryonic development normal in microgravity? Is further development and reproduction normal after return to Earth? On the other hand, does microgravity affect the organs in adult animals? Does microgravity affect the regeneration of organs? Fertilization in space is clearly demonstrated. However, subsequent embryonic development appears to be altered in microgravity. In Pleurodeles, abnormalities such as cortical cytoplasmic movements, decrease of cell adhesion, and loss of cells were observed. Although, early development was not strictly normal as a consequence of embryological regulation phenomena, young hatching larvae had normal morphological phenotypes and swimming behavior. After landing, no differences were observed between born-in-space animals to standard ones during the embryonic development to adulthood. The analyses of their offspring demonstrated that the percentages of fertilization and development were in accordance with the control animals. No genetic abnormalities were detected during the analysis of the offspring. The development of their progenies were also without characteristic differences compared to control Pleurodeles. Microgravity seems to have effects on the morphological and histological structures of organs of flight adults. However, as was the case in several experiments the number of analyzed adults was low, and it is too early to conclude on specific effects of microgravity. Moreover, in certain flights the temperature was not regulated, and an increase in temperature occurred. Conditions of these space flights had certainly influenced the samples, and consequently the interpretations of results. Space flights have clear effects on organs in regeneration. But more specifically, they have long term effects that last several weeks after the return of the animals to Earth. A similar result was also obtained for otoconia several months after landing. So far, however, no clear hypothesis could be proposed to interpret these observations.

Animals↗
PubMed1996

Secondary particle contribution to LET spectra on LDEF.

Four experiments utilizing passive detectors (P0006, P0004, A0015, M0004) were flown on LDEF to study the radiation environment. These experiments have been summarized in a companion paper (Benton et al., 1996). One of the experimental goals was to measure LET spectra at different locations and shielding depths with plastic nuclear track detectors (PNTD). It was found that the LET spectra extended well above the LET cutoff imposed by the geomagnetic field on GCR particle penetration into LEO. The high LET particles detected were mostly short-range (range < 2000 m), indicating that they were secondaries produced locally within the PNTD. The presence of these high LET particle fluences is important for the determination of dose equivalent because of the high Quality Factors (Q) involved. A relatively small fraction of particle fluence can contribute a large fraction of dose equivalent. Short-range, inelastic secondary particles produced by trapped protons in the South Atlantic Anomaly (SAA) were found to be a major contributor to the LET spectra above 100 keV/micrometer. The LET spectra were found to extend beyond the approximately 137 keV/micrometer relativistic GCR Fe peak to over 1000 keV/micrometer. The high LET tail of the LET spectra was measured in CR-39 and polycarbonate PNTDs using different techniques. GCR made a relatively modest contribution to the LET spectra as compared to the contributions from short-range secondary particles and stopping protons. LET spectra intercomparisons were made between LDEF measurements and exposures to 154 MeV accelerated proton beams. The similarities support the role of nuclear interactions by trapped protons as the major source of secondary particles in the PNTDs. Also techniques were employed to reduce the range cutoff for detection of the short-range secondaries to approximately 1 micrometer, so that essentially all secondary particles were included in the LET spectra. This has allowed a more realistic assessment of secondary contribution to dose equivalent. Comparisons of measured and calculated LET spectra have been made that demonstrate the need for more accurate modeling of secondary particles in radiation transport codes. Comparisons include preliminary calculations in which attempts have been made to include secondary particles.

Atlantic Ocean↗
PubMed1997

The determination of ice composition with instruments on cometary landers.

The determination of the composition of materials that make up comets is essential in trying to understand the origin of these primitive objects. The ices especially could be made in several different astrophysical settings including the solar nebula, protosatellite nebulae of the giant planets, and giant molecular clouds that predate the formation of the solar system. Each of these environments makes different ices with different composition. In order to understand the origin of comets, one needs to determine the composition of each of the ice phases. For example, it is of interest to know that comets contain carbon monoxide, CO, but it is much more important to know how much of it is a pure solid phase, is trapped in clathrate hydrates, or is adsorbed on amorphous water ice. In addition, knowledge of the isotopic composition of the constituents will help determine the process that formed the compounds. Finally, it is important to understand the bulk elemental composition of the nucleus. When these data are compared with solar abundances, they put strong constraints on the macro-scale processes that formed the comet. A differential scanning calorimeter (DSC) and an evolved gas analyzer (EGA) will make the necessary association between molecular constituents and their host phases. This combination of instruments takes a small (tens of mg) sample of the comet and slowly heats it in a sealed oven. As the temperature is raised, the DSC precisely measures the heat required, and delivers the gases to the EGA. Changes in the heat required to raise the temperature at a controlled rate are used to identify phase transitions, e.g., crystallization of amorphous ice or melting of hexagonal ice, and the EGA correlates the gases released with the phase transition. The EGA consists of two mass spectrometers run in tandem. The first mass spectrometer is a magnetic-sector ion-momentum analyzer (MAG), and the second is an electrostatic time-of-flight analyzer (TOF). The TOF acts as a detector for the MAG and serves to resolve ambiguities between fragments of similar mass such as CO and N2. Because most of the compounds of interest for the volatile ices are simple, a gas chromatograph is not needed and thus more integration time is available to determine isotopic ratios. A gamma-ray spectrometer (GRS) will determine the elemental abundances of the bulk cometary material by determining the flux of gamma rays produced from the interaction of the cometary material with cosmic ray produced neutrons. Because the gamma rays can penetrate a distance of several tens of centimeters a large volume of material is analyzed. The measured composition is, therefore, much more likely to be representative of the bulk comet than a very small sample that might have lost some of its volatiles. Making these measurements on a lander offers substantial advantages over trying to address similar objectives from an orbiter. For example, an orbiter instrument can determine the presence and isotopic composition of CO in the cometary coma, but only a lander can determine the phase(s) in which the CO is located and separately determine the isotopic composition of each reservoir of CO. The bulk composition of the nucleus might be constrained from separate orbiter analyses of dust and gas in the coma, but the result will be very model dependent, as the ratio of gas to dust in the comet will vary and will not necessarily be equal to the bulk value.

Ammonia↗
PubMed1999

Airborne mass spectrometers: four decades of atmospheric and space research at the Air Force research laboratory.

Mass spectrometry is a versatile research tool that has proved to be extremely useful for exploring the fundamental nature of the earth's atmosphere and ionosphere and in helping to solve operational problems facing the Air Force and the Department of Defense. In the past 40 years, our research group at the Air Force Research Laboratory has flown quadrupole mass spectrometers of many designs on nearly 100 sounding rockets, nine satellites, three Space Shuttles and many missions of high-altitude research aircraft and balloons. We have also used our instruments in ground-based investigations of rocket and jet engine exhaust, combustion chemistry and microwave breakdown chemistry. This paper is a review of the instrumentation and techniques needed for space research, a summary of the results from many of the experiments, and an introduction to the broad field of atmospheric and space mass spectrometry in general.

Atmosphere↗
PubMed2000

Potential interstellar molecules. Formation of neutral C(6)CO from C(6)CO(-*) in the gas phase.

Computations at the RCCSD(T)/aug-cc-pVDZ//B3LYP/6-31G* level of theory indicate that neutral C(6)CO is a stable species. The ground state of this neutral is the singlet cumulene oxide :C=C=C=C=C=C=C=O. The adiabatic electron affinity and dipole moment of singlet C(6)CO are 2.47 eV and 4.13 D, respectively, at this level of theory. The anion (C(6)CO)-* should be a possible precursor to this neutral. It has been formed by an unequivocal synthesis in the ion source of a mass spectrometer by the S(N)2(Si) reaction between (CH(3))(3)Si-C(triple bond)C-C(triple bond)C-C(triple bond)C-CO-CMe(3) and F(-) to form (-)C(triple bond)C-C(triple bond)C-C(triple bond)C-CO-Me(3) which loses Me(3)C* in the source to form C(6)CO(-)*. Charge stripping of this anion by vertical Franck-Condon oxidation forms C(6)CO, characterised by the neutralisation-reionisation spectrum (-NR(+)) of C(6)CO(-*), which is stable during the timeframe of this experiment (10(-6) s).

Anions↗
PubMed2000

Peptide bond formation in gas-phase ion/molecule reactions of amino acids: a novel proposal for the synthesis of prebiotic oligopeptides.

There is a general fascination with regard to the origin of life on Earth. There is an intriguing possibility that prebiotic precursors of life occurred in the interstellar space and were then transported to the early Earth by comets, asteroids and meteorites. It is probable that some part of the prebiotic molecules may have been generated by gas-phase ion/molecule reactions. Here we show experimentally that gaseous ion/molecule reactions of the amino acids, Glu and Met, may promote the synthesis of protonated dipeptides such as (Glu-Glu)H(+) and (Glu-Met)H(+) and their chemical growth to larger protonated peptides.

Amino Acids↗
PubMed2001

"Liquidless" cell staining by dye diffusion from gels and analysis by laser scanning cytometry: potential application at microgravity conditions in space.

BACKGROUND: Conventional staining of cells or tissue sections on microscope slides involves immersing the slides into solutions of dyes then rinsing to remove the unbound dye. There are instances, however, when use of stain solutions is undesirable-e.g., at microgravity conditions in space, where the possibility of accidental spill (many dyes are known carcinogens) introduces health hazard. Likewise, transporting bulk of liquid stains and rinses may be burdensome in certain situations such as field expeditions or combat. METHODS: The "liquidless" staining procedure is proposed in which the dyes are contained in thin strips of hydrated polyacrylamide or gelatin gels that have been presoaked in the stain solutions. Fluorochromes that have affinity to DNA (propidium iodide, PI; 4,6-diamidino-2-phenylindole, DAPI, Hoechst 33342) or to protein (sulforhodamine 101) were used to saturate the gels. The gel strips were placed over the prefixed cells or tissue sections deposited on microscope slides and relatively low (20 g/cm2) pressure was applied to ensure the contact. The cells were also stained by using commercially available mounting media into which DAPI or PI were admixed. Intensity of fluorescence of the PI stained cells was measured by laser scanning cytometry (LSC). RESULTS: Satisfactory cell and tissue staining, with minimal background, was achieved after 10-20 min contact between the cells and gels. Optimal concentrations of the dyes in the solutions used to presoak the gels was found to be 2-4-fold higher than the concentrations used routinely in cytometry. The measurements of intensity of cellular fluorescence by LSC revealed that the staining of DNA was stoichiometric as reflected by the characteristic cellular DNA content frequency histograms with distinct G1, S, and G2/M cell populations and 2:1 ratio of G2/M to G1 peak fluorescence. Individual gels can be saturated with more than a single dye-e.g., to obtain differential DNA and protein staining. Cell staining with DAPI or PI in the gelatin-based mounting media led to high fluorescence background while staining with DAPI in "aqueous" medium was satisfactory. CONCLUSIONS: Relatively fast staining of cells or tissue sections on microscope slides can be achieved by nonconvective dye diffusion using hydrated gels permeated with the dyes, applied to cells at low pressure. The quality of the staining provided by this methodology is comparable to conventional cell staining in dye solutions.

Coloring Agents↗