Introduction: Seeing Spores From Bacteria With an Polarized Microscope!
Hello, do you ever wanted to see spores of bacteria with a Polarizing lens that can be added to a 40 to 1000 x magnifying Microscope? Well in this Instructable I will show you how to do that. Here some videos that explain how a Polarizer lens works first of all.
Here another tutorial on Polarization lenses.
For 30 dollars the US you can get a good lens and a holder that allows you to put the Lens underneath the light source to amplify the light through the two lenses.
Carbonyl fushol stain with 5 percent Phenol.
Crystal violet in ethanol 1 percent.
Loafer Methylene blue in ethanol 1 percent.
Warning: Wear eye goggles, and gloves when handling Carbonol Fushol it has 5 percent Phenol which can be toxic (very) if a lot of it gets absorbed through the skin. Also, wear gloves for Loafer methylene blue and the crystal violet since both are toxic.
Never work with any bacteria that could be "Pathogenic" it is very dangerous and Illegal.
Step 1: Dangerous Use of Chemicals and Hotplate.
You are dealing with bacteria that could trigger reactions (extremely rare)! Again most bacteria from yogurt samples, etc can be done safely. The exception is if you have a weakened immune system.
Keep in mind that in Carbonyl fushol there is 5 percent or 4.55 percent Phenol which is very dangerous (it interferes with Protein synthesis and cause free radical cell damage and death.) The bottle reduces the chances of this chemical causing serious harm, however, they're still a risk. Also, read the MSDS it contains a dye that can cause cancer with repeated exposure- Limit your exposure with proper PPE. Phenol is rapidly absorbed so Nitrile or better gloves are NEEDED. DO NOT USE LATEX as if the phenol drop gets on you it can absorb into the skin.
Malachite green as a simple dye stain at 1-2 percent if used is toxic and must be handled responsibly by bioremediation of yeast and sent to the eco-center. Other dyes should NOT be dumped down the drain.
Step 2: Setup for the Lens.
Here a Lens that I bought for the Microscope. With gloves put it into the device that holds the lens and places it over the light output. Please see the first video for more info.
Place the lens here info on it under the microscope light source (It will be brighter, but that will be Ok).
You need a 50 to 52 mm lens with a 50-52 mm lens holder to protect the lens.
Step 3: Hot Plate and Slide Setup.
Here I am growing cultures of bacillus Lentamorphus from Soil and culturing Lactobacillus R (Non-sporing type) to compare the difference.
With a liquid sample and gloves place on an O ring clamp the slide 5 cm to 7 cm away from the hotplate. Make sure the temp does not go past 130 deg C on the slide. Use a thermometer if needed or digital thermometer to ensure the slide does not go past 130 or it could break the slide or burn it.
This method is baking bacteria with a drop of dye. Carbonyl fushol, Loafer Methylene blue, Crystal violet, etc are used as basic dyes. One drop on the culture.
For each sample 400 ml of bacteria (Bacillus had) 20 g cocoa protein mix, 20 g sugar, and 4 g Urea. The culture for lactobacillus was in 50 ml tube (5 g milk powder, 1.5 g Urea and the rest water with 1 drop dye (methylene blue and topped off with water).
Attach the coverslip while wearing gloves and let it heat up for 45 minutes and cool for 30-60 minutes afterward so the coverslip does not come off.
Step 4: Oil Immersion, and Slides on 100x Magnification.
With the slides put them under the Microscope with the new Polarization Lens. Make sure the lens is quite clear and not colored. Some models are colored, you want a clear one for this. (Important INFO.).
Start at 4 x 10 magnification and when you see something at 400x, then put either Canola oil on the slide or cedar oil on the slide and observe what you see focused at 1000x. Kimwipes a special type of material is needed to clean off with alcohol (ethanol, etc) the excess Oil.
Step 5: Final Conclusions to Polarizing Lens in Microscopes.
Well, Any basic lens with a 100x times 10 or 1000X mag with Oil immersion or greater and if you add a Polarizer lens can allow you to see spores and bacteria a lot better than a normal microscope. Also, it is easier to do since the kit can only cost you 30 dollars US from China which is quite cheap and comes with three lenses at 49 to 53 mm diameter.
I had fun playing around with this Polarization lens indeed.