RESUMO
New discoveries and ideas often occur at the confluence of events and technologies that allow them to happen. So it was with the first electron microscopic observations of individual atoms at the University of Chicago laboratory of Albert Crewe forty years ago. This paper will describe the technologies developed then, present some of the historical instrumental details and describe the rationale for the designs that came about in that laboratory over a period of about a decade.
Assuntos
Microscopia Eletrônica de Transmissão e Varredura/história , Microscopia Eletrônica de Transmissão e Varredura/instrumentação , Chicago , História do Século XX , Laboratórios , Microscopia Eletrônica de Transmissão e Varredura/métodos , UniversidadesRESUMO
Research into the neuromechanical basis of behavior, either in biomechanics, neuroethology, or neuroscience, is frequently limited by methods of data collection. Two of the most pressing needs are for methods with which to (1) record from multiple neurons or muscles simultaneously and (2) perform this recording in intact, behaving animals. In this paper we present the fabrication and testing of flexible multielectrode arrays (fMEAs) that move us significantly towards these goals. The fMEAs were used to record the activity of several distinct units in the coxa of the cockroach Blaberus discoidalis. The devices fabricated here address the first goal in two ways: (1) their flexibility allows them to be inserted into an animal and guided through internal tissues in order to access distinct groups of neurons and muscles and (2) their recording site geometry has been tuned to suit the anatomy under study, yielding multichannel spike waveforms that are easily separable under conditions of spike overlap. The flexible nature of the devices simultaneously addresses the second goal, in that it is less likely to interfere with the natural movement of the animal.
Assuntos
Extremidades/fisiologia , Insetos/fisiologia , Microeletrodos , Músculos/fisiologia , Algoritmos , Amplificadores Eletrônicos , Animais , Fenômenos Biomecânicos , Baratas , Eletrodos Implantados , Eletrofisiologia/estatística & dados numéricos , Locomoção/fisiologia , Músculos/inervação , Nanotecnologia , Polímeros , Corrida/fisiologia , XilenosRESUMO
Model silicon intracortical probes with microfluidic channels were fabricated and tested to examine the feasibility of using diffusion-mediated delivery to deliver therapeutic agents into the volume of tissue exhibiting reactive responses to implanted devices. Three-dimensional probe structures with microfluidic channels were fabricated using surface micromachining and deep reactive ion etching (DRIE) techniques. In vitro functional tests of devices were performed using fluorescence microscopy to record the transient release of Texas Red labeled transferrin (TR-transferrin) and dextran (TR-dextran) from the microchannels into 1% w/v agarose gel. In vivo performance was characterized by inserting devices loaded with TR-transferrin into the premotor cortex of adult male rats. Brain sections were imaged using confocal microscopy. Diffusion of TR-transferrin into the extracellular space and uptake by cells up to 400 microm from the implantation site was observed in brain slices taken 1 h postinsertion. The reactive tissue volume, as indicated by the presence of phosphorylated mitogen-activated protein kinases (MAPKs), was characterized using immunohistochemistry and confocal microscopy. The reactive tissue volume extended 600, 800, and 400 microm radially from the implantation site at 1 h, 24 h, and 6 weeks following insertion, respectively. These results indicate that diffusion-mediated delivery can be part of an effective intervention strategy for the treatment of reactive tissue responses around chronically implanted intracortical probes.
Assuntos
Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Eletrodos Implantados/efeitos adversos , Reação a Corpo Estranho/patologia , Reação a Corpo Estranho/prevenção & controle , Bombas de Infusão Implantáveis , Microfluídica/instrumentação , Animais , Desenho de Equipamento , Falha de Equipamento , Estudos de Viabilidade , Microeletrodos/efeitos adversos , Microfluídica/métodos , Ratos , Transferrina/administração & dosagemRESUMO
Neuronal cell networks have been reconstructed on planar microelectrode arrays (MEAs) from dissociated hippocampal pyramidal neurons. Microcontact printing (microCP) and a photoresist-liftoff method were used to selectively localize poly-L-lysine (PLL) on the surface of MEAs. Haptotaxis led to the organization of the neurons into networks localized adjacent to microelectrodes. Various grids of PLL with 2-25-microm-wide lines spaced by 50-200 microm with 15-25-microm nodes at intersection points were used to guide cell body attachment and neurite outgrowth. Bursting activity with spike amplitude attenuation was observed, and multichannel recordings detected instances of coincident firing activity. Finally, we present here an extracellular recording from a approximately 2 microm bundle of guided neurites.