BIO 171 Michelles Lab Notebooks - Portage Learning

BIO 171 Michelles Lab Notebooks - Portage Learning. Procedure: 1. This is where each step of a given protocol is recorded. 2. Be sure to clearly indicate any deviations that may occur during the experiment. Notes: Additional (helpful) information placed here General Lab Safety  Never Eat or drink in the lab o Contamination risks  Use Personal Protective Equipment (PPE) o Latex or thermal gloves (application dependent) -Cold gloves are thermal glove, thermal glove for heat o Eyewear protection o Lab coat  Never leave the lab while wearing PPE o Public hallway o Bathroom o Cafeteria Results: A summary of the final outcome of the experiment should be described here. Lab 2 Notebook Back to Home Page Title: MT02 Basics of Microscopy Objective: To understand basic components of a light microscope and how to use microscopy in a lab setting, also how to load samples into the microscope for viewing. Procedure: 1. Review parts and components that comprise a light microscope 2. Load sample with the glass slide into the microscope 3. Choose which magnification to use 4. Make necessary adjustments to optimize sample visualization Notes: Magnifying glass and microscope are similar, it helps make the object larger. Three things used for magnifying glasses are: 1. Needs lens to magnify object 2. Need an object to examine 3. Need a light source Microscopy 1. Many different types of microscope 2. Light microscope a. Uses a standard halogen lightbulb for illumination source 3. Parts a. Eyepiece i. Ocular 1. Move eyepiece apart or together to fit the width of your eyes ii. Need to use both eyes iii. You want to see 1 circle when looking into the eye piece to see a single image 1. If you see 2 circles, you have to adjust the piece, a. Either expanding or narrowing them iv. Own magnification, multiply with the power of objective to find the total magnification of the sample you are viewing 1. The lab microscope had a eyepiece of 10x b. Arm/Neck i. Thick curvature in the back of the microscope ii. Used for handling the microscope or moving it 1. Moving the microscope one hand on the arm and the other hand on the bottom of the base to carry c. Objectives i. Lens that provide part of the magnification to the microscope ii. Different microscopes have different set ups d. Revolving nosepiece i. Specific microscope used in the lab has as revolving set of lens with 4 different types or powers or magnification, it helps you increase zoom 1. Shortest objective is 4X, this magnifies the object by 4 times , also color red 2. 10X , magnifies the object by 10 times, yellow color 3. 40X, object by 40 times, color blue 4. 100X, object by 100 times, white color 5. When you hear a click it means the rotating is in place e. Stage i. Flat surface 1. A place ot put your sample f. Stage controls i. Different microscopes have different ways of holding the sample into place ii. Clamp holder 1. Pinch the two metal bars located next to the stage which opens the arms on the stage and then you push the glass slide on and slowly release, two metal brackets to hold your slide in place g. Stage clips i. Another type of stage control ii. Lift up and gently place onto the slide to hold the slide in place 1. Put into your glass cover slip to ensure your sample is not moving around h. Course/fine focus i. Outer ring on the knob – course focus, it helps make large changes in your focus ii. Inner ring on the knob, fine focus, make small changes in your focus , helps make it clearer i. Iris Diaphragm i. Piece over the light source that you can move back and forth, to controls how much light you are allowing into the sample j. Light source i. Light microscope, standard halogen lightbulb k. Base i. Where the microscope sets ii. At all times, you want the base to be steady and not rocking or vibrating which can affect your imaging 4. Power on in the back of the microscope with an adjustment knob, in this case it was on the left of the microscope which will dim or brighten your light a. Two different ways to control the light i. Roll the adjustment knob – increase intensity ii. Use the iris diaphragm – restricts light source 5. Stage guide a. One knob control vertical axis i. Top knob which brings glass slide into light path b. Other knob controls horizontal axis i. Lower knob which moves glass slide left or right c. It helps the sample just right above the light source 6. Visualization a. Types of objective – always good to note when approaching microscope what type of objectives are present i. DRY- 4x, 10x, and 40x are the dry objectives on the microscope 1. No other medium is needed to view an object, you can simply load the sample and view immediately 2. If you try to use a dry objective with oil, everything will be blurry and out of focus ii. Oil – 100x objective 1. With higher magnification, not always the case, it says oil on the side, so oil is required for imaging 2. Oil gives you a better resolution on high magnification a. You would have to slide glass coverslip out of frame, add a drop of immersion oil and with the light refraction, it can give you better image qualities b. If you try to use oil objective without oil, your sample will not look clear b. Intensity of light source- greatly impact how well you can visualize a sample i. Too bright= saturation 1. The light is so bright that you cannot see a lightly stained or opaque cell wall ii. Too dark= low visibility 1. Low visibility that you may not see your sample c. [Power objective] x [power of eyepiece] = total magnification i. Example- 15mm in diameter, using 40x objective and 5x eyepiece, the cell will now appear to the eye 200 times larger (200x) or 3000 mm in diameter d. Best way to start viewing your object is to start about midway with some light power, and having the diaphragm completely open to let un as much light source and then you can tailor it from there e. Usually working cells or tissues so we can usually start at a higher magnification but if you are ever uncertain start at the lowest power mag. f. To begin, control the course knob to roll either away or raise your sample or roll towards you to lower your sample, best to start at lowest end, make sure you do not hit the glass slide with the objective as soon as you start – slowly raise your object and while raising that stage you can look into your eyepiece