# Team:Aachen/Interlab Study/Hardware

(Difference between revisions)
 Revision as of 22:46, 22 August 2014 (view source)Mjoppich (Talk | contribs) (→Fluorescence)← Older edit Revision as of 22:57, 22 August 2014 (view source)Mjoppich (Talk | contribs) (→Fluorescence)Newer edit → Line 42: Line 42: == Fluorescence == == Fluorescence == + {{Team:Aachen/Figure|Aachen_gfp_ecoli_plate_1.jpg|center|title=Figure 1|subtitle=E. coli plate fluorescence|width=400px}} + When measuring fluorescence, two approaches can be followed + * measure the light intensity sent to fluorescence protein, measure returning light intensity and compare + * measure two samples, and detect the increase in light returned + which follows either an absolute or relative quantification. For most tasks however the relative increase in fluorescence is needed, and therefore this approach is also used for this device. + ''Figure 2'' shows the schematic principle of the fluorescence measurement with our device. - {{Team:Aachen/Figure|Aachen_gfp_ecoli_plate_1.jpg|center|title=Figure 1|subtitle=E. coli plate fluorescence|width=400px}} + {{Team:Aachen/Figure|Aachen_gfp_ecoli_plate_1.jpg|center|title=Figure 2|subtitle=Schematics of fluorescence measurement|width=300px}} === Characteristic Curve === === Characteristic Curve === + + An important observation for fluorescence measurement is that the linearity of the measurement highly depends on the chosen light sensor. + If the light sensor is not linear over the spectrum of measurement, linearity would be needed to be corrected by software. Fortunately the characteristic curve of the TSL 235 R shows perfect linearity such that no software correction is needed. + + This needed to be confirmed by measurements. We used an E. coli dilution series to perform this experiment. + ''Figure 3'' shows the recorded values. The linearity clearly can be seen, the regression coefficient of 0.99 also supports this. == Optical Density == == Optical Density ==

# Open Source DIY Hardware

Being in the measurement track and having a team of highly motivated engineering and computer science students, we tackled the challenge to build, document and evaluate our open source hardware approach.

For our daily tasks in the lab, two key devices were detected: fluorometer and OD-meter. As we use GFP most of the time, the fluorometer is designed to work best with GFP. For modularity reasons, and re-usability, it is designed such that a change to another fluorescence protein is easy.

Besides the mandatory $\mu$-controller architecture, we worked together with the Fablab Aachen to construct the device. There we have the chance to use laser cutters and 3D printers.

The core component for detecting the light intensity is the cuvette holder. Please find the 3D model we printed below:

This cuvette holder can be used for both devices: the whole in the bottom is used for fluorescence measurement, the two opposite wholes are used for the light sensor and the LED for optical density measurement respectively.

### Hardware Requirements

The key ingredients for our combined optical density/fluorescence (OD/F) device then are:

• Arduino UNO R3 (or equivalent)
• breadboard
• bluetooth Modem optional
• cuvette holder stl file download
• TSL 235 R light to frequency sensor
• LED for optical density (for 600nm we recommend: DIALIGHT - 550-2505F )
• LED for fluorescence (any 450nm blue works for iLOV, 480nm for wtGFP )

The case is from acrylic glass. The construction plan can be downloaded from insert link here.

## Fluorescence

When measuring fluorescence, two approaches can be followed

• measure the light intensity sent to fluorescence protein, measure returning light intensity and compare
• measure two samples, and detect the increase in light returned

which follows either an absolute or relative quantification. For most tasks however the relative increase in fluorescence is needed, and therefore this approach is also used for this device. Figure 2 shows the schematic principle of the fluorescence measurement with our device.

### Characteristic Curve

An important observation for fluorescence measurement is that the linearity of the measurement highly depends on the chosen light sensor. If the light sensor is not linear over the spectrum of measurement, linearity would be needed to be corrected by software. Fortunately the characteristic curve of the TSL 235 R shows perfect linearity such that no software correction is needed.

This needed to be confirmed by measurements. We used an E. coli dilution series to perform this experiment. Figure 3 shows the recorded values. The linearity clearly can be seen, the regression coefficient of 0.99 also supports this.

## Optical Density

Some general information