Alina Moreno - Post 1

Hello everyone!

My name is Alina, and I'm a senior at USU majoring in biochemistry. I have been working in Saarmaan's lab at USU for 2 years, primarily on insectary maintenance and some genetic testing. 

Now that I'm a RaHP VEC intern, I'm going to the Salt Lake City Mosquito Abatement District (SLCMAD) once a week to learn what they do, including surveillance field work. However, the main thing I'm working on with SLCMAD is attempting to develop a new diagnostic tool for Lysinibacillus sphaericus (Lsph) resistance in Culex Pipiens.

⍺-glucosidase is an important receptor in Lsph toxicity. Our hypothesis is that resistant Culex Pipiens have a lower ⍺-glucosidase activity than susceptible. We are using 4th instar larval samples and an ⍺-glucosidase activity assay kit to test this. So far, I have conducted 2 bioassays.

I want to share in this post  what the bioassays look like:

We grind the larva in PBS. 

We prepare a master mix using the kit's reagents. We add the mastermix and the dissolved samples to a 96-well plate, and we also include a well of just water and one of the calibrator included in the kit.

After leaving the plate in the spectrophotometer for 20 minutes, this is what the plate looks like. The color intensity is proportional to the activity.

The spectrophotometer gives us the data in the form of absorbance at 405 nm. To calculate activity in U/L, we use the following formula: 

⍺-Glucosidase Activity (units/L)= (((A405)final- (A405)initial)/((A405)calibrator-(A405)water))* 250 units/L