INPUT ARTICLE: Article: Use a pipette to flood the bacteria sample with several drops of crystal violet dye, sometimes called gentian violet. Wait thirty to sixty seconds. Crystal violet (CV) dissociates in aqueous solutions into CV+ and chloride (Cl) ions. These ions penetrate through the cell wall and cell membrane of both gram-positive and gram-negative cells. The CV+ ion interacts with negatively charged components of bacterial cells to stain the cells purple. Many laboratories use "Hucker's" crystal violet, which adds ammonium oxalate to prevent precipitation. Tilt the slide and use a wash bottle to squirt a small stream of distilled or tap water over the top of the slide. The water should run down over the surface of the smear, but not be aimed directly at it. Do not rinse excessively, which might remove the stain from Gram positive bacteria. Use a pipette to cover the smear with iodine. Let it sit for at least 60 seconds, then rinse off with the same careful method. Iodine, in the form of negatively charged ions, interacts with CV+ to form large complexes of crystal violet and iodine (CVI complexes) within the inner and outer layers of the cell. This will trap the purple crystal violet colour in the cell, wherever it has stained. Iodine is corrosive. Avoid inhalation, ingestion, or skin contact. A 1:1 mix of acetone and ethanol is typically used for this critical step, which must be timed carefully. Hold the slide at an angle, then add the decoloriser until no more violet colour is visible in the draining runoff. This typically takes under 10 seconds, or even less time if the decoloriser contains higher concentrations of acetone. Stop immediately, or the decoloriser will remove the crystal violet stain from both gram-positive and negative cells, and the stain will have to be repeated. Immediately rinse off excess decoloriser, using the earlier technique. Pure (95%+) acetone can be used instead. The more acetone there is, the faster the decoloriser will work, requiring more precise timing. If you are having trouble timing this step, consider adding the decoloriser drop by drop. A counterstain, typically safranin or fuchsin, is used to add extra contrast between gram-negative and gram-positive bacteria, by staining decolorised (gram-negative) bacteria pink or red. Leave it on for at least 45 seconds, then rinse off. Fuchsin will stain many gram-negative bacteria more intensely, such as haemophilus spp and legionella spp. This may make it a better option for beginners. You may allow the slide to air dry, or blot it dry using bibulous paper sold for this purpose. The Gram stain is complete.

SUMMARY: Flood the smear with crystal violet. Rinse off the crystal violet gently. Flood the smear with iodine, then rinse. Add a decoloriser, then rinse quickly. Flood the smear with counterstain, then rinse. Dry the slide.

INPUT ARTICLE: Article: In order to reach the lock slider for push or pull locks, you will need to straighten the hanger out, then bend it again in the right manner to reach the it. Depending on your vehicle, this may require a number of bends or few at all. You may need to guess and check the proper bends in the coat hanger a few times in order to get it right. Try not to bend the hanger too far, as bends will reduce its ability to press or pull directly on the slider. If you need to pull the slider to unlock it in your vehicle, you will want to bend the end of the hanger into a hook that can snag the ledge on it. If you need to push it, it's more important that you bend the tip in such a manner that can put the force necessary on the slider to push it over. Use a pair of pliers to make a small, strong hook or ledge. If you are sliding the hanger in the gap in the door frame instead of a window gap, it will need to be very thin. If you are unable to pry the window away from the door frame, you will need to instead slide the hanger through the very narrow gap between the door itself and the frame. The door and door frame are separated by a thin layer of weather proofing that you may damage in the process. Press the hanger through the weather proof layer and into the car. Some vehicles will not permit the hanger to pass through because of the angle. You will then have to find a way to pry the window away from the door. If you tear the weather proofing, you will need to replace it or the door may leak. Once the tip of the hanger is inside the cabin of the car, you may not be able to see the slide for the door lock from your position. Ask a friend to stand on the passenger side of the car and guide you by looking through the window. You can still find the lock without a second pair of eyes, but enlisting some help will make the process far easier. Be patient and continue trying until you are able to snag or press the slide. Once you get the hanger on the slide, push or pull it as necessary to unlock the door. This may take a number of tries and you also may need to make changes to the bends in the coat hanger in order to get to the slide from the right angle. Be patient and continue trying until you are able to snag or press the slide. Be careful not to do further damage to the weather proofing as you move the hanger around.

SUMMARY:
Straighten out the hanger and bend it again as necessary. Bend the hook into a short hook or ledge to push or pull the slide. Slide the hanger between the door and door frame. Have a friend guide you from the passenger side. Press or pull on the slide to unlock the door.