The separation of affinity flocculated yeast cell debris using a pilot-plant scroll decanter centrifuge

The use of a scroll decanter centrifuge for the removal and dewatering of affinity-flocculated yeast cell debris from a crude homogenate is described. Laboratory shear modulus measurements were used to compare the structure of flocculated and nonflocculated sediments and to indicate the dewatering conditions under which the sediment could be discharged from the centrifuge. The structure of the flocculated sediment was such that a dry beach could be used within the centrifuge while still being able to discharge the solids. The scroll decanter performance for recovery and dewatering of the flocculated homogenate was found to be independent of feed flow rate and differential scroll rate. Eighty-five percent of the solid material was recovered from the flocculated homogenate while the extent of sediment dewatering resulted in the loss of only 7% of the soluble protein in the sediment. The supernatant clarity matched that achieved by low-gravity laboratory centrifugation studies.     

Density gradient fractionation by piston displacement.

A method for the fractionation of centrifuged density gradients is described. A piston forced into the centrifuge tube from above displaces the gradient, while its tip collects liquid from the small volume immediately below it without disturbing the unfractionated part of the gradient. The centrifuge tube holder permits sensitive visual detection of bands of light scattering particles which may then be rapidly and precisely extracted. Continuous fractionation of the entire gradient is also possible.     

Medical monitoring during the NASA Artificial Gravity-Bed Rest Pilot Study

The NASA artificial gravity-bed rest pilot study (AGPS) was designed to investigate the efficacy of daily exposure to a +Gz acceleration gradient for counteracting the physiologic decrements induced by prolonged bed rest. Test subjects were continuously monitored by a physician for signs and symptoms of pre-syncope, motion sickness, and arrhythmias while on the centrifuge. In this article, we have summarized the medical monitoring observations that were made during the AGPS and included an assessment of the relative usefulness of the information provided by the various monitoring tools in making a decision to terminate a centrifuge spin.     

A comparison of two centrifuge techniques for constructing vulnerability curves: insight into the ‘open‐vessel’ artifact

A vulnerability curve (VC) describes the extent of xylem cavitation resistance. Centrifuges have been used to generate VCs for decades via static‐ and flow‐centrifuge methods. Recently, the validity of the centrifuge techniques has been questioned. Researchers have hypothesized that the centrifuge techniques might yield unreliable VCs due to the open‐vessel artifact. However, other researchers reject this hypothesis. The focus of the dispute is centered on whether exponential VCs are more reliable when the static‐centrifuge method is used rather than the flow‐centrifuge method. To further test the reliability of the centrifuge technique, two centrifuges were manufactured to simulate the static‐ and flow‐centrifuge methods. VCs of three species with open vessels of known lengths were constructed using the two centrifuges. The results showed that both centrifuge techniques produced invalid VCs for Robinia because the water flow through stems under mild tension in centrifuges led to an increasing loss of water conductivity. In addition, the injection of water in the flow‐centrifuge exacerbated the loss of water conductivity. However, both centrifuge techniques yielded reliable VCs for Prunus, regardless of the presence of open vessels in the tested samples. We conclude that centrifuge techniques can be used in species with open vessels only when the centrifuge produces a VC that matches the bench‐dehydration VC.     

Large-scale perfusion culture process for suspended mammalian cells that uses a centrifuge with multiple settling zones

A high-cell-density perfusion culture process, using a novel centrifuge, was developed. The centrifuge has spiral multiple settling zones to separate cells from culture medium. Because of the multiple zones, the separation area can be efficiently increased without enlarging the diameter of the centrifuge. The centrifuge used in this study had a separation capacity of 2600 ml culture medium min^–1 at 100g of the centrifugal force. A new cell separation and withdrawal method was also developed. The cells separated in the centrifuge can be withdrawn easily from the centrifuge with no cell clogging by feeding a liquid carrier such as a perfluorocarbon into the centrifuge and pushing the cells out with the liquid carrier. By this culture process, monoclonal antibodies were produced with mouse-human hybridoma X87X at a cell density of about 8 × 10⁶ cells ml^–1 for 25 days. This centrifuge culture shows promise as a large-scale perfusion culture process.     

Immobilization of endoplasm flowing contiguous to the actin cables upon electrical stimulus inNitella internodes

Summary Using a stroboscopic centrifuge microscope, we demonstrated that, when actin cables (=bundles of F-actin) had been previously removed locally from the cell cortex ofNitella internodes, the passively flowing endoplasm found there under centrifugal force did not stop at all upon electrical stimulus, while the actively flowing endoplasm contiguous to the actin cables at the normal cell cortex promptly stopped following the stimulus and was immobilized for several seconds. The results present evidence that, upon electrical stimulus, the presence of actin cables is required to immobilize the endoplasm flowing contiguous to the actin cables in a state that resists displacement by centrifugal force.Abbreviations APW artificial pond water - CMS centrifuge microscope     

Aquatic animals and use of centrifuge in space experiments]

There are some experiments which do not need a centrifuge apparatus in space, however, for other space experiments the use of centrifuge is indispensable as the control. The characteristics of these two types of space experiments are explained. Generally, the more quantitatively the phenomenon is analyzed, the greater the need of centrifuge apparatus becomes. The centrifuge in space can realize various gravity values ranging from 0 to 1 G, and this will extend the present biology to a more universal biology.     

Centrifuge Modeling of Cadmium Migration in Saturated and Unsaturated Soils

Researchers recently have come to recognize that geotechnical centrifuge can provide a powerful experimental tool for investigating the flow and transport of inorganic contaminants in soils. Inert, non-adsorptive inorganic compounds (e.g., sodium ions) rather than adsorptive heavy metals are often used as the chemical for the investigation of pollutant transport behavior in most centrifuge modeling. To overcome the shortcomings of this approach, centrifuge tests for the study of one-dimensional pollutant migration in both saturated and unsaturated soils were designed using an adsorptive pollutant (i.e., cadmium) and conducted at two centrifugal accelerations. In this study, the concentration profile of adsorptive cadmium along the depth of soil, the moisture content varying with the soil depth and the transport behavior of the cadmium in soils were investigated. The centrifuge results show that the cadmium concentration profiles are found to be similar for the centrifuge models performed at 15 g and 20 g and the one-dimensional moisture movement in the unsaturated soil can be reproducible. The validity of centrifuge modeling of adsorptive pollutants might be affected by the g-level that, in turn, determines the centrifuge testing time and affects the sorption equilibrium.     

Gravitropism in Phycomyces: threshold determination on a clinostat centrifuge

The absolute sensitivity of sporangiophores of Phycomyces blakesleeanus to centrifugal acceleration was determined on a clinostat centrifuge. The centrifuge provides centrifugal accelerations ranging from 10(-4) to 6 x g. The rotor of the centrifuge, which accommodates 96 culture vials with single sporangiophores, is clinostatted, that is, turning "head over", at slow speed (1 rev min(-1)) while it is running. The negative gravitropism of sporangiophores is characterized by two components: a polar angle, which is measured in the plane of bending, and an aiming-error angle, which indicates the deviation of the plane of bending from the vector of the centrifugal acceleration. Dose-response curves were generated for both angles with centrifugations lasting 3, 5, and 8 h. The threshold for the polar angle depends on the presence of statoliths, so-called octahedral protein crystals in the vacuoles. The albino strain C171 carAcarR (with crystals) has a threshold near 10(-2) x g while the albino strain C2 carAgeo-3 (without crystals) has a threshold of about 2 x 10(-1) x g. The threshold for the aiming error angle is ill defined and is between 10(-2) and 10(-1) x g. The threshold for the polar angle of the wild type NRRL 1555 (with crystals) is near 8 x 10(-2) x g.     

Total containment of a continuous-flow zonal centrifuge

A continuous-flow, high throughput, turbine-driven zonal centrifuge has been contained for use with biologically hazardous materials. The centrifuge and all ancillary equipment are enclosed in three sealed cabinets that are maintained at negative pressure, decontaminated with ethylene oxide, and provide cooling for the process fluids. Sample handling is semiautomated and remotely controlled for ease of operation and includes an automated decontamination system. The centrifuge subsystems can be decontaminated prior to an engineer servicing the machine, and apparatus is provided for absolute filtration of the centrifuge turbine exhaust.     

The scale-down of an industrial disc stack centrifuge

The means are described whereby a disc stack centrifuge may be scaled-down by up to 10-fold of its separation capacity. The centrifuge separation characteristics so measured are suitable for direct scale-up predictions of centrifuge performance where only small volumes of particle suspension are available for study. Such an ability to scale-down is especially important in the processing of biological particles where for example, in the early stage of process development, there is often insufficient fermentation broth for fullscale studies. Scale-down is achieved by the reduction of the number of discs available for separation purposes and by the careful positioning of these discs in the overall disc stack. A combination of dye tracer and particle separation studies are used to optimise the disc stack configuration. The resulting grade efficiency curve is an accurate reflection of the curve for the full-scale centrifuge especially in the critical design region specifying centrifuge throughput for near complete particle recovery.     

Tube inserts as aids in preparative ultracentrifugation

Conventional preparative ultracentrifugation was modified by using inserts in the centrifuge tubes, causing a reduction of the time necessary to centrifuge a protein at reduced rotor velocity from suspension.     

Evaluation of a class III biological safety cabinet for enclosure of an ultracentrifuge

An evaluation of a special safety cabinet housing a high-speed centrifuge was made. The cabinet enclosed both the top access port and the drive and pumping machinery of the centrifuge. A titanium rotor was loaded with tubes containing a bacterial culture, weakened, and driven until rotor rupture occurred. There were several bent and broken components in the centrifuge, and bacteria leaked from the vacuum chamber. Although the forces were sufficient to displace the cabinet, none of the test bacteria were found outside the cabinet.     

The total containment of a batch-type zonal centrifuge.

A batch-type zonal centrifuge has been modified and totally contained for use with biologically hazardous materials. A sealed cabinet encloses the centrifuge and the ancilliary equipment. It is operated with a flow of filtered air when the zonal system is on, decontaminated with ethylene oxide, and maintained at a negative pressure throughout. The centrifuge subsystems can be drained, flushed, and decontaminated with ethylene oxide before an engineer services the machine. The sample handling system within the cabinet is remotely controlled.     

Separation efficiency and particle size distribution in relation to manure type and storage conditions

Mechanical separation of liquid animal manure can be an effective technique for production of a liquid and a nutrient-rich solid fraction. The efficiency of separators depends on the physical and chemical composition of the animal manure. Therefore, the particle size composition was measured for different types of manure before treatment with a decanting centrifuge and a screw press. Storage of pig manure reduces the total dry matter content, and the content of small particles (< 0.0016 mm) is reduced more than the content of large particles. In consequence, the proportion of large particles will increase, while the portion of small particles will decrease. The separation efficiency of the screw press was found to be low, as this separator only retains particles > 1 mm. The decanter centrifuge retained all the particles > 0.02 mm and was therefore much more efficient than the screw press. Separation efficiency was also found to be highly dependent on the type of manure used.     

Visualization of a rapid,red/far-red light-dependent reaction by centrifuge microscopy

Summary Using a centrifuge microscope with stroboscopic illumination, we examined the effects of light irradiation on the passive movement of chloroplasts in dark-adapted mesophyll cells ofVallisneria gigantea. While irradiation with red light accelerates the passive gliding of chloroplasts produced by centrifugal force, irradiation with far-red light negates this effect. Irradiation with blue light does not accelerate the passive gliding, while red light is completely effective even in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea, an inhibitor of photosynthesis. An apparently active movement of chloroplasts can be induced by irradiation with red or blue light only in the presence of the far-red light-absorbing form of phytochrome. The significance of the reaction in the light with respect to the regulation of cytoplasmic streaming is discussed.Abbreviations APW artificial pond water - CMS centrifuge microscope of the stroboscopic type - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - Pfr phytochrome, far-red light-absorbing form - Pr phytochrome, red light-absorbing form     

The use of laboratory centrifugation studies to predict performance of industrial machines: studies of shear-insensitive and shear-sensitive materials

A method for using a bench-top centrifuge is described in order to mimic the recovery performance of an industrial-scale centrifuge, in this case a continuous-flow disc stack separator. Recovery performance was determined for polyvinyl acetate particles and for biological process streams of yeast cell debris and protein precipitates. Recovery of polyvinyl acetate particles was found to be well predicted for these robust particles. The laboratory centrifugation scale-down technique again predicted the performance of the disc stack centrifuge for the recovery of yeast cell debris particles although there was some suggestion of over-prediction at high levels of debris recovery due to the nature of any cell debris aggregates present. The laboratory centrifuge scale-down technique also proved to be an important investigative probe into the extent of shear-induced breakup of shear-sensitive protein precipitate aggregates during recovery in continuous high speed centrifuges. Such breakup can lead to over 10-fold reduction in separator capacity.     

8. Tube Inserts as Aids in Preparative Ultracentrifugation

ABSTRACT

Conventional preparative ultracentrifugation was modified by using inserts in the centrifuge tubes, causing a reduction of the time necessary to centrifuge a protein at reduced rotor velocity from suspension.     

Tube Inserts as Aids in Preparative Ultracentrifugation

Abstract

Conventional preparative ultra centrifugation was modified by using inserts in the centrifuge tubes, causing a reduction of the time necessary to centrifuge a protein at reduced rotor velocity from suspension.     

Feasibility Study of Using Centrifuge for Investigating LNAPL Migration in Unsaturated Soils

Centrifuge modeling appears potentially useful for studying geo-environmental problems such as pollutant migration in subsurface systems. In this study, the “modeling of models” technique was used to validate the feasibility of using a geotechnical centrifuge to model the transport behavior of light non-aqueous phase liquids (LNAPLs) in unsaturated soils. All the experiments were conducted to simulate a gasoline spill from a leaking underground storage tank (UST) and the subsequent subsurface migration of the gasoline. When the gravity in the centrifuge reached the desired g-level, the gasoline was released from the UST and then it migrated in the unsaturated soil corresponding to a prototype time equivalent of one year. After the centrifuge tests, soil samples were collected using sampling tubes and the concentrations of individual constituent in the LNAPL were directly measured by means of gas chromatograph analysis. Results obtained from the centrifuge tests at different g-levels show that similar migration patterns are found for LNAPL transport in unsaturated porous media. The location of the peak concentration and the behavior of lateral spreading can be adequately described. In addition, centrifuge test data show that the migration pattern of LNAPLs is related to the soil type and the physical properties of individual constituents in the LNAPLs.