Transcript

Firstly spot-on 5 microliters of a polysaccharide, in solution onto a nitrocellulose sheet that has been divided into 1-centimetre squared squares. In this video different commercial polysaccharides were used with the dilution of 1 to 5 going down the nitrocellulose. This method can be used to determine how soil it is to plant root mucilage, fungal mucilage or bacterial mucilage. Different dilutions also can be used for the gelatinous polysaccharides that you may have.

To start you need to cut a small section of the tip of your pipette to aid pipetting. Once complete, cover the nitrocellulose sheet and leave to air-dry for a couple of hours. When the nitrocellulose is dried, re-dampen the sheet with deionized water. The soil used to cover the nitrocellulose must be free of any larger particles and sterilised. After layering the nitrocellulose with soil, cover the sheet and air-dry overnight.

Gently lift the nitrocellulose sheet to remove excess soil. Then submerge the sheet that has been incubated overnight into approximately 120 millilitres of deionized water for one second and then remove. Repeat this step once more for around one second. After the washes, place a sheet into a fresh weighing boat, cover and then air dry for a couple more hours.

To scan, place the sheet face down onto a flatbed scanner. ImageJ was used to quantify the soil. In order to quantify the plot, a 1-centimetre line will be required to set the scale. You can use that from their nitrocellulose which has the lines drawn on it already to set the scale. Click on analyse and then set the scale.

After the scale is set, draw a circle around one of the dots at the top of the nitrocellulose dot, note the area size as this has to be the same for all the other dots. After the dot is selected click on analyse and then set measurements ensuring the mean grey value is ticked. After checking values, close the toolbar then press on analyse measure. Collect all the values and open them on the spreadsheet. Remove the background values from the results to produce your mean grey values for your dots. Please note that the larger mean grey values, the more soil has adhered to the nitrocellulose. This figure shows the final output you will get.