Team:Newcastle/Project/shape shifting

From 2013.igem.org

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==Our Project==
==Our Project==
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We aim to put L-forms into a single cell wide microfluidics channels (that are as small as 1 μm wide) with differently shaped end chambers. We will then observe whether the L-forms will grow and take up the shape of these chambers due to their lack of cell wall.
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Most of the bacteria has evolved to have a cell wall - a rigid structure which protects the bacteria from the variety of enviromental hazards such as mechanical stress, osmotic rupture and lysis. The cell wall often sxerves as a docking point to many proteins including various receptors and adherence sites. Along with these properties cell wall provides the cell with a rigid boundary and helps bacteria to acquire and preserve their shape. In ''Bacillus subtilis'' along with other proteins, a group of proteins termed Penicillin Binding Proteins (pbp), usually anchored in the cell wall, is involved in the formation of the rod shape. When the cells lose their cell wall they automatically lose these proteins to the environment as they are being made. The cells lose the support and turn into a sphere as it is the most energetically favourable state (ratio of surface area to volume is minimal, and membrane curvature is more-or-less constant). However it has been previously observed that these cells can become elongated and 'squeeze' into the channels with a smaller diameter than theirs.
====Microfluidics====
====Microfluidics====
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Microfluidics are a manipulation of fluid in the micro domain. The microfluidics chambers were designed using autoCAD, software used for computer-aided design and drafting, to produce silicon wafer master moulds. Directed flow within the microfluidic wafer will physically manoeuvre L-form cells into the designed chambers, where they will be maintained by nutrient media. This will enable single cell level analysis for L-forms.
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Microfluidics are a manipulation of fluid in the micro domain. The microfluidics chambers were designed using autoCAD, software used for computer-aided design and drafting, to produce silicon wafer master moulds. Directed flow within the microfluidic wafer will physically maneuver L-form cells into the designed chambers, where they will be maintained by nutrient media. This will enable single cell level analysis for L-forms.
====Visualization of Cell Membrane====
====Visualization of Cell Membrane====
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The full description of the model can be found on this [https://2013.igem.org/Team:Newcastle/Modelling/CellShapeModel page].
The full description of the model can be found on this [https://2013.igem.org/Team:Newcastle/Modelling/CellShapeModel page].
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==Experiments==
 
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==Results==
 

Revision as of 10:24, 24 September 2013

 
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Contents

Shape Shifting

Our Project

Most of the bacteria has evolved to have a cell wall - a rigid structure which protects the bacteria from the variety of enviromental hazards such as mechanical stress, osmotic rupture and lysis. The cell wall often sxerves as a docking point to many proteins including various receptors and adherence sites. Along with these properties cell wall provides the cell with a rigid boundary and helps bacteria to acquire and preserve their shape. In Bacillus subtilis along with other proteins, a group of proteins termed Penicillin Binding Proteins (pbp), usually anchored in the cell wall, is involved in the formation of the rod shape. When the cells lose their cell wall they automatically lose these proteins to the environment as they are being made. The cells lose the support and turn into a sphere as it is the most energetically favourable state (ratio of surface area to volume is minimal, and membrane curvature is more-or-less constant). However it has been previously observed that these cells can become elongated and 'squeeze' into the channels with a smaller diameter than theirs.

Microfluidics

Microfluidics are a manipulation of fluid in the micro domain. The microfluidics chambers were designed using autoCAD, software used for computer-aided design and drafting, to produce silicon wafer master moulds. Directed flow within the microfluidic wafer will physically maneuver L-form cells into the designed chambers, where they will be maintained by nutrient media. This will enable single cell level analysis for L-forms.

Visualization of Cell Membrane

We would also be able to visualise the shape of the cell membrane and confirm our hypothesis by using FM.595 membrane stain to stain the membrane red. This will show that the shape of L-forms can be easily manipulated.

News

8th August

The rough model outline is now ready. Microfluidics chamber is currently in design.

5th August

Follow-up meeting with Dr.David Swailes. During the meeting, the concepts of the first, and most primitive model were identified. Maths added.

30th July

First meeting with Dr.David Swailes, head of fluid mechanics department in the school of Mechanical Engineering at Newcastle University. During the meeting, the underlying physiology of the growth was identified and agreed upon. Based on the physiology, the approach towards the modelling was decided to have three parts: Unrestricted growth, Turning Point and Restricted growth and thus change of the shape.

Modelling

For the purposes of the study the complex model of the growing cell inside of the confined space can be broken down to simpler models of the system at three phases. The first phase would be a constantly growing cell, followed by a model of the cell gradually adopting the shape of the boundaries.

The full description of the model can be found on this page.