Limits...
Dual-compartment neurofluidic system for electrophysiological measurements in physically segregated and functionally connected neuronal cell culture.

Kanagasabapathi TT, Ciliberti D, Martinoia S, Wadman WJ, Decré MM - Front Neuroeng (2011)

Bottom Line: Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated.The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining.In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques.

View Article: PubMed Central - PubMed

Affiliation: Minimally Invasive Healthcare Department, Philips Research Laboratories Eindhoven, Netherlands.

ABSTRACT
We developed a dual-compartment neurofluidic system with inter-connecting microchannels to connect neurons from their respective compartments, placed on a planar microelectrode arrays. The design and development of the compartmented microfluidic device for neuronal cell culture, protocol for sustaining long-term cultures, and neurite growth through microchannels in such a closed compartment device are presented. Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated. The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining. In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques. Bidirectional signal propagation between the compartments is demonstrated using functional connectivity maps. CC analysis and connectivity maps demonstrate that the two neuronal populations are not only functionally connected within each compartment but also with each other and a well connected functional network was formed between the compartments despite the physical barrier introduced by the microchannels.

No MeSH data available.


Related in: MedlinePlus

Statistical analysis of correlation on multiple devices; box plots of intra and inter-compartmental correlation levels, divided per region, in 17 devices (statistical parameters used in the two-step analysis: “median” for both the steps).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3198030&req=5

Figure 6: Statistical analysis of correlation on multiple devices; box plots of intra and inter-compartmental correlation levels, divided per region, in 17 devices (statistical parameters used in the two-step analysis: “median” for both the steps).

Mentions: Box plots of correlation levels obtained from original spike trains in the same compartment (“Intra”) and the other compartment (“Inter”) as shown in Figure 6 in comparison to the surrogate spike train in the other compartment (“Inter_Shuffled”) reveal very clearly that a genuine inter-compartmental connectivity is present in the considered devices. A paired sign test on “Inter” and “Inter_Shuffled” of each region confirmed the existence of a significant difference between such values for each of the four identified regions of electrodes (p < 0.01). The above mentioned box plots do not show a clear fall off of the inter-compartmental connectivity with the distance from the barrier of microchannels. The Friedman’s test on the four groups of “Inter” values revealed that they are not statistically different from each other (p = 0.1843). This high degree of connectivity may be explained by the fact that cortical neurons usually develop long neurites.


Dual-compartment neurofluidic system for electrophysiological measurements in physically segregated and functionally connected neuronal cell culture.

Kanagasabapathi TT, Ciliberti D, Martinoia S, Wadman WJ, Decré MM - Front Neuroeng (2011)

Statistical analysis of correlation on multiple devices; box plots of intra and inter-compartmental correlation levels, divided per region, in 17 devices (statistical parameters used in the two-step analysis: “median” for both the steps).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3198030&req=5

Figure 6: Statistical analysis of correlation on multiple devices; box plots of intra and inter-compartmental correlation levels, divided per region, in 17 devices (statistical parameters used in the two-step analysis: “median” for both the steps).
Mentions: Box plots of correlation levels obtained from original spike trains in the same compartment (“Intra”) and the other compartment (“Inter”) as shown in Figure 6 in comparison to the surrogate spike train in the other compartment (“Inter_Shuffled”) reveal very clearly that a genuine inter-compartmental connectivity is present in the considered devices. A paired sign test on “Inter” and “Inter_Shuffled” of each region confirmed the existence of a significant difference between such values for each of the four identified regions of electrodes (p < 0.01). The above mentioned box plots do not show a clear fall off of the inter-compartmental connectivity with the distance from the barrier of microchannels. The Friedman’s test on the four groups of “Inter” values revealed that they are not statistically different from each other (p = 0.1843). This high degree of connectivity may be explained by the fact that cortical neurons usually develop long neurites.

Bottom Line: Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated.The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining.In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques.

View Article: PubMed Central - PubMed

Affiliation: Minimally Invasive Healthcare Department, Philips Research Laboratories Eindhoven, Netherlands.

ABSTRACT
We developed a dual-compartment neurofluidic system with inter-connecting microchannels to connect neurons from their respective compartments, placed on a planar microelectrode arrays. The design and development of the compartmented microfluidic device for neuronal cell culture, protocol for sustaining long-term cultures, and neurite growth through microchannels in such a closed compartment device are presented. Using electrophysiological measurements of spontaneous network activity in the compartments and selective pharmacological manipulation of cells in one compartment, the biological origin of network activity and the fluidic isolation between the compartments are demonstrated. The connectivity between neuronal populations via the microchannels and the crossing-over of neurites are verified using transfection experiments and immunofluorescence staining. In addition to the neurite cross-over to the adjacent compartment, functional connectivity between cells in both the compartments is verified using cross-correlation (CC) based techniques. Bidirectional signal propagation between the compartments is demonstrated using functional connectivity maps. CC analysis and connectivity maps demonstrate that the two neuronal populations are not only functionally connected within each compartment but also with each other and a well connected functional network was formed between the compartments despite the physical barrier introduced by the microchannels.

No MeSH data available.


Related in: MedlinePlus