An apparatus for anesthetizing small laboratory animals.

A simple apparatus was designed for simultaneous general anesthesia of a number of small laboratory animals with spontaneous respiration. Anesthesia is consistent over many hr. The apparatus consists of an oxygen tank and vaporizer, a glass distribution bottle, and a non-return circuit of separate inspiratory and expiratory valves for each animal. The equipment is inexpensive to construct and can be rapidly adjusted for exact control of gas flow and anesthetic depth.     

An automatic, closed-circuit oxygen consumption apparatus for small animals.

An apparatus suitable for the continuous measurement of oxygen consumption of rats and mice is described. The system uses a motorized syringe dispenser to deliver fixed volumes of oxygen to a closed animal chamber. The dispenser is controlled by a micro-differential pressure switch to maintain chamber pressure slightly above ambient. The rate of oxygen consumption is determined by timing the interval between successive operations of the dispenser. The system has proved suitable for a range of experimental conditions and treatments.     

Elasticity and stress relaxation of a very small vocal fold

Across mammals many vocal sounds are produced by airflow induced vocal fold oscillation. We tested the hypothesis that stress-strain and stress-relaxation behavior of rat vocal folds can be used to predict the fundamental frequency range of the species' vocal repertoire. In a first approximation vocal fold oscillation has been modeled by the string model but it is not known whether this concept equally applies to large and small species. The shorter the vocal fold, the more the ideal string law may underestimate normal mode frequencies. To accommodate the very small size of the tissue specimen, a custom-built miniaturized tensile test apparatus was developed. Tissue properties of 6 male rat vocal folds were measured. Rat vocal folds demonstrated the typical linear stress-strain behavior in the low strain region and an exponential stress response at strains larger than about 40%. Approximating the rat's vocal fold oscillation with the string model suggests that fundamental frequencies up to about 6 kHz can be produced, which agrees with frequencies reported for audible rat vocalization. Individual differences and time-dependent changes in the tissue properties parallel findings in other species, and are interpreted as universal features of the laryngeal sound source.     

An automated apparatus for quantitative aliquot measurement of phosphorylated intermediates

An automatic dry ashing apparatus is described. It permits the colorimetric measurement of organic phosphate compounds in the nanomole range either as discrete samples or as the effluent from a column. In its present form it can be used to ash fractions up to 0.5 ml at rates up to three per minute.     

An apparatus for the measurement of lung volume and compliance in mice

Pressure-volume (P-V) curves and total lung capacity (TLC) were measured in excised lung of mice using a water manometer and a closed system in which the humidity and temperature were controlled. In pathogen-free mice there are no significant differences in elastic properties of these lungs in relation to their age. The measured TLC in those normal mice was approximately 2.9 ml. This relatively simple apparatus which allows one to make sensitive and accurate measurements of pulmonary function in mice and other small animals.     

An apparatus for olfactory isolation of small mammals for the study of chemocommunication]

Olfactory isolation of individual experimental and control animals seems to be necessary to investigate the role of chemosignals in the biology of small rodents. Each cage was isolated in a polyethylene bag and applied with purified air drawn through wash bottles containing concentrated sulphuric acid. Sulphuric acid aerosols were bound with saturated calcium chloride. Calcium chloride served also to humidify the air. Possible improvements to the apparatus are discussed.     

Quantitative measurement of starch in very small amounts of leaf tissue

Summary A specific enzyme method is described for the routine estimation of starch in small quantities (10–30 mg) of dried leaf tissue. A -glucanase-free preparation of amyloglucosidase is employed to hydrolyse starch to glucose; this is subsequently estimated by the glucose oxidase technique. The method gives result which agree closely with those obtained by a specific iodine-precipitation method.     

Optical measurement of presynaptic calcium currents.

Measurements of presynaptic calcium currents are vital to understanding the control of transmitter release. However, most presynaptic boutons in the vertebrate central nervous system are too small to allow electrical recordings of presynaptic calcium currents (I(Ca)pre). We therefore tested the possibility of measuring I(Ca)pre optically in boutons loaded with calcium-sensitive fluorophores. From a theoretical treatment of a system containing an endogenous buffer and an indicator, we determined the conditions necessary for the derivative of the stimulus-evoked change in indicator fluorescence to report I(Ca)pre accurately. Matching the calcium dissociation rates of the endogenous buffer and indicator allows the most precise optical measurements of I(Ca)pre. We tested our ability to measure I(Ca)pre in granule cells in rat cerebellar slices. The derivatives of stimulus-evoked fluorescence transients from slices loaded with the low-affinity calcium indicators magnesium green and mag-fura-5 had the same time courses and were unaffected by changes in calcium influx or indicator concentration. Thus both of these indicators were well suited to measuring I(Ca)pre. In contrast, the high-affinity indicator fura-2 distorted I(Ca)pre. The optically determined I(Ca)pre was well approximated by a Gaussian with a half-width of 650 micros at 24 degrees C and 340 micros at 34 degrees C.     

An electrohydraulic apparatus for the measurement of static and dynamic properties of rabbit muscles

The apparatus described in this communication enables the force-velocity relationship to be determined for whole rabbit muscles in vivo and their resistance to fatigue to be assessed at specified rates of external work. The ergometer generates constant-velocity motion, controlling a force of up to 50 N, over a range of velocity up to 500 mm/s and a distance of 20 mm. This distance corresponds to the range of shortening of the rabbit tibialis anterior muscle from full plantar flexion to full dorsiflexion of the foot, equivalent to approximately 28% fiber shortening. Activated muscles can be allowed to shorten at constant velocity from any point on their isometric force trajectory. Cyclic releases for fatigue testing can be made at rates up to 30 releases/min over a period of 6-8 h. The timing of the release and return strokes of the ergometer is under the control of a digital programmer that also synchronizes the delivery of activating stimuli to the muscle nerve and trigger signals to the recording equipment. An electrohydraulic design was chosen because it is simpler to engineer than an electromagnetic actuator, is reliable in continuous cyclic use, and can be assembled, at least in part, from available industrial components.     

Gramicidin-mediated currents at very low permeant ion concentrations.

Current-voltage relations have been measured for the fluxes of caesium ions through pores formed by gramicidin in lipid bilayer membranes. The ionic currents have been separated from capacitative currents using a bridge circuit with an integrator as null-detector. The conductances during brief voltage pulses were small enough to avoid the effects of diffusion polarization and the ionic strength was raised using choline chloride or magnesium sulfate to reduce the effects of double-layer polarization. Under these conditions the current-voltage relations have the same shape at 0.1 and 1 mM, but different shapes for higher concentrations. These data demonstrate that the fluxes do not obey independence for concentrations above 10 mM, but they cannot be used in isolation to support a particular value of the binding constant. The shape observed at low concentrations suggests that entry of ions into the pore remains weakly potential dependent even at 300 mV.     

A controlled-temperature apparatus for measurement of hydrogen peroxide production.

An apparatus is described which permits continuous assays of hydrogen peroxide production from biological materials, including whole cells. Temperature in the assay mixture is controlled, for work above or below room temperatures. Data obtained by use of the apparatus are presented, as illustration of the effects of temperature upon peroxide output.     

Adaptive processing techniques based on hidden Markov models for characterizing very small channel currents buried in noise and deterministic interferences.

Techniques for characterizing very small single-channel currents buried in background noise are described and tested on simulated data to give confidence when applied to real data. Single channel currents are represented as a discrete-time, finite-state, homogeneous, Markov process, and the noise that obscures the signal is assumed to be white and Gaussian. The various signal model parameters, such as the Markov state levels and transition probabilities, are unknown. In addition to white Gaussian noise, the signal can be corrupted by deterministic interferences of known form but unknown parameters, such as the sinusoidal disturbance stemming from AC interference and a drift of the base line owing to a slow development of liquid-junction potentials. To characterize the signal buried in such stochastic and deterministic interferences, the problem is first formulated in the framework of a Hidden Markov Model and then the Expectation Maximization algorithm is applied to obtain the maximum likelihood estimates of the model parameters (state levels, transition probabilities), signals, and the parameters of the deterministic disturbances. Using fictitious channel currents embedded in the idealized noise, we first show that the signal processing technique is capable of characterizing the signal characteristics quite accurately even when the amplitude of currents is as small as 5-10 fA. The statistics of the signal estimated from the processing technique include the amplitude, mean open and closed duration, open-time and closed-time histograms, probability of dwell-time and the transition probability matrix. With a periodic interference composed, for example, of 50 Hz and 100 Hz components, or a linear drift of the baseline added to the segment containing channel currents and white noise, the parameters of the deterministic interference, such as the amplitude and phase of the sinusoidal wave, or the rate of linear drift, as well as all the relevant statistics of the signal, are accurately estimated with the algorithm we propose. Also, if the frequencies of the periodic interference are unknown, they can be accurately estimated. Finally, we provide a technique by which channel currents originating from the sum of two or more independent single channels are decomposed so that each process can be separately characterized. This process is also formulated as a Hidden Markov Model problem and solved by applying the Expectation Maximization algorithm. The scheme relies on the fact that the transition matrix of the summed Markov process can be construed as a tensor product of the transition matrices of individual processes.