Improved fluorescent probes for the measurement of rapid changes in membrane potential

To improve the quality of fluorescent voltage-sensitive probes twenty new styryl dyes were synthesized. Some of the new probes are significantly better than any used in the past. A signal-to-noise ratio of 90 root mean square (rms) noise was obtained for an optical recording of action potentials from neuroblastoma cells maintained in monolayer culture. The fluorescence fractional change of the optical signal is as large as 14%/100 mV. Photodynamic damage and bleaching are much less significant with the new probes. These fluorescent probes can be used to measure small and rapid changes in membrane potential from single cells maintained in monolayer cultures, from single cells in invertebrate ganglia, from their arborization, and from other preparations. The optical measurement can be made with a standard fluorescent microscope equipped with DC mercury illumination. Guidelines for the design of even better fluorescent probes and more efficient instruments are suggested

Temporally-structured acquisition of multidimensional optical imaging data facilitates visualization of elusive cortical representations in the behaving monkey

Fundamental understanding of higher cognitive functions can greatly benefit from imaging of cortical activity with high spatiotemporal resolution in the behaving non-human primate.To achieve rapid imaging of high-resolution dynamicsof cortical representations of spontaneous and evoked activity ,we designed a novel data acquisition protocol for sensory stimulation by rapidly interleaving multiple stimuli in continuous sessions of optical imaging with voltage-sensitive dyes. We also tested a new algorithm for the “temporally structured componentanalysis” (TSCA) of a multidimensional timeseries that was developed for our new data acquisitionprotocol, but was tested only on simulated data (Blumenfeld, 2010). In addition to the raw data, the algorithm incorporates prior knowledge about the temporal structure of the data as well as input from other information. Here we showed thatTSCA can successfully separate functional signal components from other signals referred to as noise. Imaging of responses to multiple visual stimuli, utilizing voltage-sensitive dyes, wasperformed on the visual cortex of awake monkeys. Multiple cortical representations,including orientation and ocular dominance maps as well as thehitherto elusive retinotopic representation of orientation stimuli, were extracted in only 10 secondsof imaging, approximately two orders of magnitude faster than accomplished by conventional methods. Since the approach is rather general, other imaging techniques may also benefit from the same stimulation protocol. This methodology can thus facilitate rapid optical imaging explorations in monkeys, rodents and other specieswith a versatility and speed that were not feasible before

Voltage-sensitive dyes imaging of neocortical activity

Neural computations underlying sensory perception, cognition, and motor control are performed by populations of neurons at different anatomical and temporal scales. Few techniques are currently available for exploring the dynamics of local and large range populations. Voltage-sensitive dye imaging (VSDI), based on organic voltage probes, reveals neural population activity in areas ranging from a few tens of micrometers to a couple of centimeters, or two areas up to ~10 cm apart. VSDI provides a submillisecond temporal resolution and a spatial resolution of ~50 µm. The dye signal emphasizes subthreshold synaptic potentials. VSDI has been applied in the mouse, rat, gerbil, ferret, tree shrew, cat, and monkey cortices to explore the lateral spread of retinotopic or somatotopic activation; the dynamic spatiotemporal pattern resulting from sensory activation, including the somatosensory, olfactory, auditory, and visual modalities; and motor preparation and the properties of spontaneously occurring population activity. In this introduction, we focus on VSDI in vivo and review results obtained mostly in the visual system in our laboratory

Optical recording of synaptic potentials from processes of single neurons using intracellular potentiometric dyes.

To record post synaptic potentials or electrical activity from processes of single cells in a central nervous system (CNS) preparation in situ, voltage sensitive dyes can be injected intracellularly, thereby staining only the cell under investigation. We report the structure, evaluation, and synthesis of 11 fluorescent styryl dyes developed for iontophoretic injection. The optical signals that represent small synaptic potentials from single processes of iontophoretically injected cells are expected to be very small and, therefore, such measurements are not easy. We report the methodology that permitted the optical recording of action potentials from a 3-micron axon and the recording of small synaptic potentials from the processes of single cells in the segmental ganglia of the leech. The same dyes also proved useful for optical recording of action potentials of anterogradely labeled axons, following local extracellular injection at a remote site in a mammalian CNS preparation

Interhemispheric Synchrony of Spontaneous Cortical States at the Cortical Column Level

In cat early visual cortex, neural activity patterns resembling evoked orientation maps emerge spontaneously under anesthesia. To test if such patterns are synchronized between hemispheres, we performed bilateral imaging in anesthetized cats using a new improved voltage-sensitive dye. We observed map-like activity patterns spanning early visual cortex in both hemispheres simultaneously. Patterns virtually identical to maps associated with the cardinal and oblique orientations emerged as leading principal components of the spontaneous fluctuations, and the strength of transient orientation states was correlated with their duration, providing evidence that these maps are transiently attracting states. A neural mass model we developed reproduced the dynamics of both smooth and abrupt orientation state transitions observed experimentally. The model suggests that map-like activity arises from slow modulations in spontaneous firing in conjunction with interplay between excitation and inhibition. Our results highlight the efficiency and functional precision of interhemispheric connectivity.status: publishe