How to Prepare a Simple Stain
Clean your slide., Dry the slide with either paper towels or lens paper. , Draw a target circle on the back side of the slide., Place a drop of water on to the target circle: This water is going to hold your organisms on the slide , Flame the...
Step-by-Step Guide
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Step 1: Clean your slide.
To clean the slide you can use tap water, make sure that the water is not too hot or on high because it will cause the slide to break. -
Step 2: Dry the slide with either paper towels or lens paper.
, Use a blue sharpie for this because the color will show more.
This circle will be used as a target to place your organisms on and also know where to look at on the microscope, hence its name, target circle. ,, Hold the loop at the end and place the loop part into the fire.
You want the loop to red hot which will let the loop become sterile. , Hold the loop for a few seconds to cool, if the loop is too hot you will burn your bacteria.
Do not swing the loop around to cool it or you will burn someone around you , Placing the loop on the organisms you want and GENTLY rub the organism around on the surface to get it on the loop.
Do not get too much or it will be too thick to see under the microscope ,, Flaming the loop again is help keep it sterile, let is cool again and set to the side , Using your slide holder, grab the slide and pass it through the flame several times allowing the heat-fix process to occur.
The Heat fixing process kills any bacteria that may be present and causes the cell stick to the slide.
Do not over-heat the slide or the cell can become distorted or unclear under the microscope , Still holding the slide, let it cool for about a 30 seconds to a minute , The gloves will protect you from the dye that will be used to stain the slide , This will allows the excess stain you put on the run off into the sink.
It will make a big mess in the sink , Try to cover the whole target circle with the stain, but don’t have too much of the stain or it will be too hard to observe under the microscope. ,, Using the squirting bottle of water aim at a 45 degree angle starting at the top of where the smear begin to squirt your water.
It must be at this degree so then you will not wash away the organisms you stained. , It must be done gently or you might break your slide , -
Step 3: Draw a target circle on the back side of the slide.
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Step 4: Place a drop of water on to the target circle: This water is going to hold your organisms on the slide
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Step 5: Flame the inoculating loop.
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Step 6: Let the loop cool.
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Step 7: Take a loopful or your organism.
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Step 8: Mix the organisms in the water on the slide: spread the organisms and the water around the barrier of the target circle until it is a thin sheet of water covering the circle
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Step 9: Flame the inoculating loop again.
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Step 10: Heat fix the slide.
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Step 11: Allow the slide to cool.
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Step 12: Put gloves on now to continue to the next step.
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Step 13: Place the staining tray on the corner of the sink.
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Step 14: Cover the smear with a few drops of Methylene blue.
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Step 15: Wait 30- 60 seconds to let the stain dye the smear.
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Step 16: Grasp the slide with the slide holder and rinse.
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Step 17: Gently blot dry with the bibulous paper: You can place the slide in between the pages of the bibulous paper and gently push down on the booklet.
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Step 18: Ready to observe under the microscope: You can see the organisms under objectives 10x and 40x
Detailed Guide
To clean the slide you can use tap water, make sure that the water is not too hot or on high because it will cause the slide to break.
, Use a blue sharpie for this because the color will show more.
This circle will be used as a target to place your organisms on and also know where to look at on the microscope, hence its name, target circle. ,, Hold the loop at the end and place the loop part into the fire.
You want the loop to red hot which will let the loop become sterile. , Hold the loop for a few seconds to cool, if the loop is too hot you will burn your bacteria.
Do not swing the loop around to cool it or you will burn someone around you , Placing the loop on the organisms you want and GENTLY rub the organism around on the surface to get it on the loop.
Do not get too much or it will be too thick to see under the microscope ,, Flaming the loop again is help keep it sterile, let is cool again and set to the side , Using your slide holder, grab the slide and pass it through the flame several times allowing the heat-fix process to occur.
The Heat fixing process kills any bacteria that may be present and causes the cell stick to the slide.
Do not over-heat the slide or the cell can become distorted or unclear under the microscope , Still holding the slide, let it cool for about a 30 seconds to a minute , The gloves will protect you from the dye that will be used to stain the slide , This will allows the excess stain you put on the run off into the sink.
It will make a big mess in the sink , Try to cover the whole target circle with the stain, but don’t have too much of the stain or it will be too hard to observe under the microscope. ,, Using the squirting bottle of water aim at a 45 degree angle starting at the top of where the smear begin to squirt your water.
It must be at this degree so then you will not wash away the organisms you stained. , It must be done gently or you might break your slide ,
About the Author
Cheryl Collins
Enthusiastic about teaching practical skills techniques through clear, step-by-step guides.
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