Latest revision as of 01:54, 22 October 2009

Motor Preliminary Trials

Before we obtained the final silicon-based motors, we did several trials to test whether the microorganisms can be attached to the surface of a membrane-based motor. This involved the following:

The micromotor was shaped into defined tiny structures.
The micromotor was chemically modified to coat the biotin on the surface.

Here we report our trials by using Immobolin-P membrane which is easy to cut and coat with biotin.

Trial 1.The fragmentation of the Immobolin-P membrane with the help of a mini-homogenizer.

The Immobolin-P membrane was first made wet and consequently homogenized with a mini-homogenizer.

Results: The membrane could not be broken into small pieces

Figure 1. mini-homogenizer cutting

Trial 2.The fragmentation of the Immobolin-P membrane with a generated shearing force by moistening and followed by vortexing

The Immobolin-P membrane was first moistened, then it was put into a 10ml centrifugation tube. The tube was then filled with glass beads, and subjected to vortexing subsequently.

Results: The membrane remained intact.

Figure 2. moistening by vortexing‎

Trial 3.The fragmentation of the Immoboin-P membrane by moistening and the use of liquefied nitrogen

The Immobolin-P membrane was moistened and liquefied nitrogen was poured onto it. We then tried to break the membrane into pieces by human hands.

Results: The membrane remained intact.

Trial 4.The biotinylation of the Immobolin-P membrane followed by the fragmentation process of the membrane by human hands

The Immobolin-P membrane was first biotinylated on one side and cut into very small pieces by human hands. Then, the membrane fragments were put into a mould made with aluminum foil and fixed into it with the help of glue. The moulds along with the membrane fragment were cut with a Leica-crytomicrotome into pieces with the size of 100µm x 60µm x 100µm.

Results: The Immobolin-P membrane was observed under microscopy(Fig.4).The sharp was not regular, and the size was around 100 um scale.

Figure 3. cutting by crytomicrotome ‎

Figure 4.the cut membrane under microscopy

Trial 5. Binding polar expression E. coli to the biotin side-coated Immobolin-P membrane The Immobolin-P membrane underwent the process of Trial 4. The diameter of it was around 100um. And one side of the membrane was coated biotin that can bind to the polar expression E.coli.

Results: Under microscopy, we can observe the polar expression E.coli(the small dots) were mostly bind to the biotin coated membrane side. And other sides also randomly binded some E.coli.

Figure 5.the membrane bound with E. coli under microscopy

@@ Line 3: / Line 3: @@
 {{Team:HKU-HKBU/header}}
-==Membrane Production==
+=Motor Preliminary Trials=
+Before we obtained the final silicon-based motors, we did several trials to test whether the microorganisms can be attached to the surface of a membrane-based motor. This involved the following:
+* The micromotor was shaped into defined tiny structures.
+* The micromotor was chemically modified to coat the biotin on the surface.
+Here we report our trials by using Immobolin-P membrane which is easy to cut and coat with biotin.
+'''Trial 1.'''The fragmentation of the Immobolin-P membrane with the help of a mini-homogenizer.
-===Step 1===
 The Immobolin-P membrane was first made wet and consequently homogenized with a mini-homogenizer.
-Results: The membrane could not be broken into small pieces.
+'''Results''': The membrane could not be broken into small pieces
-[[Image:HKU-HKBU_motor_results_1.png? | center]]
+[[Image:HKU-HKBU_motor_results_1.png| center | thumb |200px|'''Figure 1.''' mini-homogenizer cutting]]
+'''Trial 2.'''The fragmentation of the Immobolin-P membrane with a generated shearing force by moistening and followed by vortexing
+The Immobolin-P membrane was first moistened, then it was put into a 10ml
+centrifugation tube. The tube was then filled with glass beads, and subjected to vortexing subsequently.
+'''Results''': The membrane remained intact.
+[[Image:HKU-HKBU_motor_results_2.png| center | thumb |200px|'''Figure 2.''' moistening by vortexing‎]]
+'''Trial 3.'''The  fragmentation of the Immoboin-P membrane by moistening and the use of liquefied nitrogen
+The Immobolin-P membrane was moistened and liquefied nitrogen was poured onto it. We then tried to break the membrane into pieces by human hands.
+'''Results''': The membrane remained intact.
+'''Trial 4.'''The   biotinylation of the Immobolin-P membrane followed by the fragmentation process of the membrane by human hands
-===Step 2===
+The Immobolin-P membrane was first biotinylated on one side and cut into very small pieces by human hands. Then, the membrane fragments were put into a mould made with aluminum foil and fixed into it with the help of glue. The moulds along with the membrane fragment were cut with a Leica-crytomicrotome into pieces with the size of 100µm x 60µm x 100µm.
-The Immobolin-P membrane was first moistened, then it was put into a 10ml centrifugation tube. The tube was then totally filled with glass beads, and undergo vortexing subsequently.
-Results: The membrane remained intact.
+'''Results''': The Immobolin-P membrane was observed under microscopy('''Fig.4''').The sharp was not regular, and the size was around 100 um scale.
-[[Image:HKU-HKBU_motor_results_2.png? | center]]
+[[Image:HKU-HKBU_motor_results_4.png| center | thumb |200px| '''Figure 3'''. cutting by crytomicrotome ‎]]
-[[Image:HKU-HKBU_motor_results_3.png? | center]]
+[[Image:HKU-HKBU_motor_results_MVB.jpg| center|thumb|200px|'''Figure 4'''.the cut membrane under microscopy]]
-===Step 3===
-The Immobolin-P membrane was moistened and liquefied nitrogen was poured onto it. The membrane was broken into pieces by human hands.
-Results: The membrane remained intact.
-===Step 4===
+'''Trial 5.''' Binding polar expression ''E. coli'' to the biotin side-coated Immobolin-P membrane
-The Immobolin-P membrane was first biotinylated and cut into very small pieces by human hands. Then, the membrane fragments were put into a mould made with aluminium foil and fixed into it with the help of glue. The mould along with the membrane fragment were cut with a Leica-crytomicrotome into further smaller pieces the size of 100umx60umx100um.
+The Immobolin-P membrane underwent the process of '''Trial 4'''. The diameter of it was around 100um. And one side of the membrane was coated biotin that can bind to the polar expression ''E.coli''.
-[[Image:HKU-HKBU_motor_results_4.png? | center]]
+'''Results''': Under microscopy, we can observe the polar expression ''E.coli''(the small dots) were mostly bind to the biotin coated membrane side. And other sides also randomly binded some ''E.coli''.
-[[Image:HKU-HKBU_motor_results_5.png? | center]]
-[[Image:HKU-HKBU_motor_results_6.png? | center]]
-===Step 5===
+[[Image:HKU-HKBU_motor_results_MVB2.jpg| center|thumb|200px|'''Figure 5'''.the membrane bound with ''E. coli'' under microscopy]]
-Some elemental silicon fragments were put inside a 1-ml eppendorf tube. The protein-biotin complex and some concentrated HCL were then added into the same tube. The tube was put inside a water bath for 2 hours. The silicon fragments were made dry by rinsing with PBS and followed by air drying. Then, some streptavidin containing beads were added onto the fragments. The fragments were observed under a microscope.
-Results: No beads were bound to the silicon fragments.
-===Step 6===
-The silicon fragments were silanized by soaking them for 2 h in a solution of aminopropyl triethoxyl silane (3% aminopropyl triethoxyl silane, 2% acetic acid, 5% water, 90% ethanol), then rinsed with ethanol, and dried with a PCR machine for 5 mins. The amino-coated rotors (Fig. 2Bh) were then reacted with 1 mM succinimidyl-6-(biotinamido)-6-hexana- mido hexanoate (EZ-Link NHS-LC-LC-biotin; Pierce, Rock- ford, IL) dissolved in 40 mM phosphate buffer (pH 8.0) for 1 h at 37°C. Then strepatavidin beads were allowed to bind onto it and the fragments were observed under a microscope.
 {{Team:HKU-HKBU/footer}}

Team:HKU-HKBU/Motor Preliminary Trials

From 2009.igem.org

Latest revision as of 01:54, 22 October 2009

Motor Preliminary Trials