Whole-Cell Recording in the Awake Brain

Lee, Doyun; Lee, Albert K.

doi:10.1101/pdb.top087304

Cited by 3 publications

(3 citation statements)

References 40 publications

(49 reference statements)

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“…However, this study has some limitations. First, the recording time can be influenced by other factors (craniotomy quality (Lee et al, 2014 ), animal movement (Lee and Lee, 2017 ), brain pulsation (Levy et al, 2012 ), and cleanliness of the recording electrode (Hamill et al, 1981 ; Stett et al, 2003 ). These other factors cannot be controlled based on identical standards and may cause a biased result.…”

Section: Discussionmentioning

confidence: 99%

Optimal Pipette Resistance, Seal Resistance, and Zero-Current Membrane Potential for Loose Patch or Breakthrough Whole-Cell Recording in vivo

Yan

Fang

Zhang

et al. 2020

Front. Neural Circuits

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In vivo loose patch and breakthrough whole-cell recordings are useful tools for investigating the intrinsic and synaptic properties of neurons. However, the correlation among pipette resistance, seal condition, and recording time is not thoroughly clear. Presently, we investigated the recording time of different pipette resistances and seal conditions in loose patch and breakthrough whole-cell recordings. The recording time did not change with pipette resistance for loose patch recording ( R p -loose) and first increased and then decreased as seal resistance for loose patch recording ( R s -loose) increased. For a high probability of a recording time ≥30 min, the low and high cutoff values of R s -loose were 21.5 and 36 MΩ, respectively. For neurons with R s -loose values of 21.5–36 MΩ, the action potential (AP) amplitudes changed slightly 30 min after the seal. The recording time increased as seal resistance for whole-cell recording ( R s -tight) increased and the zero-current membrane potential for breakthrough whole-cell recording (MP zero-current ) decreased. For a high probability of a recording time ≥30 min, the cutoff values of R s -tight and MP zero-current were 2.35 GΩ and −53.5 mV, respectively. The area under the curve (AUC) of the MP zero-current receiver operating characteristic (ROC) curve was larger than that of the R s -tight ROC curve. For neurons with MP zero-current values ≤ −53.5 mV, the inhibitory or excitatory postsynaptic current amplitudes did not show significant changes 30 min after the seal. In neurons with R s -tight values ≥2.35 GΩ, the recording time gradually increased and then decreased as the pipette resistance for whole-cell recording ( R p -tight) increased. For the high probability of a recording time ≥30 min, the low and high cutoff values of R p -tight were 6.15 and 6.45 MΩ, respectively. Together, we concluded that the optimal R s -loose range is 21.5–36 MΩ, the optimal R p -tight range is 6.15–6.45 MΩ, and the optimal R s -tight and MP zero-current values are ≥2.35 GΩ and ≤ −53.5 mV, respectively. Compared with R s -tight, the MP zero-current value can more accurately discriminate recording times ≥30 min and <30 min.

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Section: Discussionmentioning

confidence: 99%

Optimal Pipette Resistance, Seal Resistance, and Zero-Current Membrane Potential for Loose Patch or Breakthrough Whole-Cell Recording in vivo

Yan

Fang

Zhang

et al. 2020

Front. Neural Circuits

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“…A separate approach to address the limitation of head-fixed behaviors is to improve the head-mounted physiological recording systems compatible with free behavior. Progress is being made in miniaturizing recording devices for optical 53,54 , extracellular [55][56][57] or intracellular [58][59][60][61] recordings to make them compatible with free behavior. However, the need to miniaturize the devices leads to compromises in data throughput or quality.…”

Section: Figure 1 | Comparison Between Free Behaviors Virtual Realitmentioning

confidence: 99%

An animal-actuated rotational head-fixation system for 2-photon imaging during 2-d navigation

Voigts

Harnett

2018

Preprint

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Understanding how the biology of the brain gives rise to the computations that drive behavior requires high fidelity, large scale, and subcellular measurements of neural activity. 2-photon microscopy is the primary tool that satisfies these requirements, particularly for measurements during behavior. However, this technique requires rigid head-fixation, constraining the behavioral repertoire of experimental subjects. Increasingly, complex task paradigms are being used to investigate the neural substrates of complex behaviors, including navigation of complex environments, resolving uncertainty between multiple outcomes, integrating unreliable information over time, and/or building internal models of the world. In rodents, planning and decision making processes are often expressed via head and body motion. This produces a significant limitation for head-fixed two-photon imaging. We therefore developed a system that overcomes a major problem of headfixation: the lack of rotational vestibular input. The system measures rotational strain exerted by mice on the head restraint, which consequently drives a motor, rotating the constraint system and dissipating the strain. This permits mice to rotate their heads in the azimuthal plane with negligible inertia and friction. This stable rotating head-fixation system allows mice to explore physical or virtual 2-D environments. To demonstrate the performance of our system, we conducted 2-photon GCaMP6f imaging in somas and dendrites of pyramidal neurons in mouse retrosplenial cortex. We show that the subcellular resolution of the system's 2-photon imaging is comparable to that of conventional head-fixed experiments. Additionally, this system allows the attachment of heavy instrumentation to the animal, making it possible to extend the approach to largescale electrophysiology experiments in the future. Our method enables the use of state-of-the-art imaging techniques while animals perform more complex and naturalistic behaviors than currently possible, with broad potential applications in systems neuroscience.

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“…Although they have been extensively used for in vitro studies, their use in in vivo studies has been limited due to their large form factor. The conventional intracellular recording system has significant limitations: (1) owing to the large form factor of the technologies involved, recordings have mostly been obtained from anesthetized animals with the exception of a few studies 6,7 , (2) recordings are obtained from one neuron at a time (serial recording), (3) their use requires extraordinary manual skill and tedious operations, which results in a long training period for neurophysiologists, and (4) duration of recordings obtained are typically short (45-60 min in anesthetized animals, 5-30 min in awake head-fixed animals 8,9 ), often due to mechanical disruptions at the electrode-cell interface. These challenges have impeded chronic intracellular recording studies from a population of neurons in anesthetized and awake animals.…”

Section: Introductionmentioning

confidence: 99%

Engineering microscale systems for fully autonomous intracellular neural interfaces

Kumar

Baker

Okandan³

et al. 2020

Microsyst Nanoeng

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Conventional electrodes and associated positioning systems for intracellular recording from single neurons in vitro and in vivo are large and bulky, which has largely limited their scalability. Further, acquiring successful intracellular recordings is very tedious, requiring a high degree of skill not readily achieved in a typical laboratory. We report here a robotic, MEMS-based intracellular recording system to overcome the above limitations associated with form factor, scalability, and highly skilled and tedious manual operations required for intracellular recordings. This system combines three distinct technologies: (1) novel microscale, glass-polysilicon penetrating electrode for intracellular recording; (2) electrothermal microactuators for precise microscale movement of each electrode; and (3) closed-loop control algorithm for autonomous positioning of electrode inside single neurons. Here we demonstrate the novel, fully integrated system of glass-polysilicon microelectrode, microscale actuators, and controller for autonomous intracellular recordings from single neurons in the abdominal ganglion of Aplysia californica (n = 5 cells). Consistent resting potentials (<−35 mV) and action potentials (>60 mV) were recorded after each successful penetration attempt with the controller and microactuated glass-polysilicon microelectrodes. The success rate of penetration and quality of intracellular recordings achieved using electrothermal microactuators were comparable to that of conventional positioning systems. Preliminary data from in vivo experiments in anesthetized rats show successful intracellular recordings. The MEMS-based system offers significant advantages: (1) reduction in overall size for potential use in behaving animals, (2) scalable approach to potentially realize multi-channel recordings, and (3) a viable method to fully automate measurement of intracellular recordings. This system will be evaluated in vivo in future rodent studies.

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Whole-Cell Recording in the Awake Brain

Cited by 3 publications

References 40 publications

Optimal Pipette Resistance, Seal Resistance, and Zero-Current Membrane Potential for Loose Patch or Breakthrough Whole-Cell Recording in vivo

Optimal Pipette Resistance, Seal Resistance, and Zero-Current Membrane Potential for Loose Patch or Breakthrough Whole-Cell Recording in vivo

An animal-actuated rotational head-fixation system for 2-photon imaging during 2-d navigation

Engineering microscale systems for fully autonomous intracellular neural interfaces

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