Prestained slides are prevalent in the market. These slides are coated with various solutions and are dried so that the user can perform quick and fast testing. One such slide can be found here: http://cellvu.com/products/drm-900-pre-stained-morphology-slides/. This slide is coated with a mixture of methylene blue and cresyl violet such that when blood is smeared on the slide, the white blood cells in the smear stain blue.
The task is to reverse engineer the stain coated on the prestained cell vu slide. This involves experimenting with different solvents, different concentrations of methylene blue and cresyl violet, and different drying techniques. Completion of the task will be denoted when the proper stain is uniformly dried on a slide such that when it interacts with blood, it stains white blood cells blue just as the cell vu slide does within a reasonable time (~6 minutes). This can be determined by placing a drop of blood on the slide and placing a coverslip on top of it. After 6 minutes, the white blood cells should have stained blue. The pre-stained slide should be stable/dry - once coated with your dye, it should be able to stay as such for multiple days in a dry container and then be pulled out for use when needed for White Blood Cell staining.
Some guidances that may be of use:
-A now public patent protocol/procedure about a similar dye (we haven't fully explored this yet, so it might be worth checking this out as a first try after tinkering with the Cell Vu's): https://www.google.com/patents/US3796594
Materials (other than the general lab space/equipment + Microscope which you will need, we can ship many of these to you if hired):
-Cell Vu slides for a base case/gold standard comparison
-Other chemicals and solvents you may need (ethanol etc.)
-Microscope (powerful enough to see blood cells on slide)
-Microscope Slides for coating the dye
-Lancets: we've been pricking ourselves using standard diabetes lancets for a small drop sample of blood that's placed on the slide and then squished with a cover slip. This is a relatively simple/quick/safe way to test, but if you dislike a simple needle, you can also use pre-collected EDTA coagulated blood (this will take a while to arrive though
-General lab equipment to experiment with your dye!
-Specific protocol with concentrations, chemicals, drying mechanisms, and timing (ie. leaving overnight, 6 hours etc.) of methylene blue and cresyl violet. Basically a specific procedure to replicate how you've been able to create the dye + coat it on the slide.
-Images of slide, both regular and under microscope indicating stained blood cells. Similar to the image attachment "blood_stain.jpg" below. Note this is at a low magnification (you should confirm at higher magnifications for White blood cell characteristics).
-Platelet Staining - if you can get the stain to work on Platelets along with White Blood Cells, a $2000 bonus.
-Microchannel - will explain more of this later (if applicable), $2000 bonus.