iLCD Reproduce the chemical activation of Calcium channels in - - PowerPoint PPT Presentation

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iLCD Reproduce the chemical activation of Calcium channels in - - PowerPoint PPT Presentation

Mar 17, 2024 140 likes •782 views

iGEM Valencia Lightning Cell Display iLCD Reproduce the chemical activation of Calcium channels in yeast. Design a model for the electrical activation. Construction of device to supply the initial conditions of the characterization

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iGEM Valencia Lightning Cell Display

iLCD

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Reproduce the chemical activation

  • f Calcium channels in yeast.

Design a model for the electrical activation. Construction of device to supply the initial conditions of the characterization experiments. Characterization of the system parameters.

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Luminometer

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30 μL KOH

20 40 60 80 100 50 100 150 200 Time ¡(s) Luminiscence ¡(a.u.)

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Characterization. Different volumes of KOH (μL)

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  • Controls. Alkali shock

Cch1: Calcium channel deficient mutant Mid1: Calcium channel deficient mutant WT: Wild Type EDTA: Ethylenediaminetetraacetic acid (calcium quelant) KCl: Potassium chloride

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  • Controls. Alkali shock

Cch1: Calcium channel deficient mutant Mid1: Calcium channel deficient mutant WT: Wild Type EDTA: Ethylenediaminetetraacetic acid (calcium quelant) KCl: Potassium chloride

Channel deficient mutants

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Device

Voltage amplitude: ±0’1V During time intervals from 20 ms

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4,5 Volts during 5 seconds

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4,5 Volts during 5 seconds

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  • Controls. Electrical Shock.

WT: Wild Type Cch1: Calcium channel deficient mutant Mid1: Calcium channel deficient mutant SD + Coel: Medium with no yeasts incubated with coelenterazine. SD - Coel: Medium with yeasts incubated without coelenterazine. EDTA: Ethylenediaminetetraacetic acid (calcium quelant)

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Most important parameters

  • Quantity of light emission
  • Reproduibility
  • Refreshing time
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Characterization. Different time, same voltage

1s 5s 10s

Dead yeasts :(

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4,5 Volts during 5 seconds

WT: Wild Type WT - coe: Wild Type incubated without coelenterazine

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Device

Voltage amplitude ±0’1V During time intervals from 20 ms 96 outputs

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VIDEO JUNY

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VIDEO JUNY

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Reproduce the chemical activation

  • f Calcium channels in yeast.

Design a model for the electrical activation of these channels which points to a feasible activation at acceptable voltage levels. Construction of a homemade device to supply, very precisely, the initial conditions for the experiments. Mesurements and precise characterization

  • f LECs (Lighting Emiting Cell)
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Reproduce the chemical activation

  • f Calcium channels in yeast.

Design a model for the electrical activation of these channels which points to a feasible activation at acceptable voltage levels. Construction of a homemade device to supply, very precisely, the initial conditions for the experiments. Mesurements and precise characterization

  • f LECs (Lighting Emiting Cell)

Construction of a labview program which allows the cooperative work of several cultures to build the iLCD!!!

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Achievements:

  • Electrical induction for a desired cell behaviour
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  • Characterization and construction of the

first bio-screen of the history

Achievements:

  • Electrical induction for a desired cell behaviour
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  • Characterization and construction of the

first bio-screen of the history

Achievements:

  • BBa_K222000 (Aequorin) + Full characterization
  • Electrical induction for a desired cell behaviour
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  • Human Practices Report

First review of iGEM and Human Practices (2005-2009) Biggest survey ever made (1288 people)

  • Characterization and construction of the

first bio-screen of the history

Achievements:

  • BBa_K222000 (Aequorin) + Full characterization
  • Electrical induction for a desired cell behaviour
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