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Lab 1. Introduction to Microscopy and the Cell

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David Ruckle

on 8 January 2019

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Transcript of Lab 1. Introduction to Microscopy and the Cell

Lab 1.
Introduction to Microscopy and the Cell

- To learn the appropriate and careful usage of a compound light microscope
- Calibration an ocular micrometer
- Wet Mount and Staining Live Specimen
- Observe and distinguish the cellular characteristics of prokaryotes and eukaryotes
Microscope Parts
- When you first get to lab:

- Learn the various parts of the microscope (pg 12: Figure 1)

- TAs will test and confirm your knowledge on the various microscope parts

- TAs will Sign you in when you are ready to move on to the other exercises
TA Demo:
Proper Usage of Microscope
How to carry Microscope properly

Guidelines on proper microscope use (pg. 13)
1) Lower stage all the way down at 4X before placing/replacing slide
2) Coarse focus only for 4x objective
3) Fine focus on 10X, 40X objectives
4) Do not skip objectives
5) Move the nosepiece back to 4x and lower the stage all the way down before removing the slide

- used to reveal specific components of the cell
- Neutral Red (cytoplasm), Janus Green B (mitochondria), Iodine (amyloplasts), Methylene Blue (nuclei)
Final Remarks
- work in groups of 2
- make sure to gain good understanding on microscope use (it will be on the lab final)
- clean up area and get sign-out signature from a TA
- make sure you have completed the 7 FOVs
- paste all necessary components into a lab notebook by next week
- Purpose of calibration:
estimate the distance across one ocular space to use as ruler to measure

- Ocular micrometers are limited.

- Ask TA to get you an ocular micrometer

Calibration of Ocular Micrometer
Wet Mount:
- for live specimen in aqueous environment
- place a drop of environmental solution on top of the live specimen
Wet Mount and Staining
TA demo:
Prepare wet mount
Put 1 drop of stain on side of coverslip
Use kimwipe to draw stain by capillary action
TA will Demo in Lab
Staining by wicking
TA Demo:
Wet Mount
1) Place dry specimen on slide.
2) Place 1 drop of Protoslo
3) Put 1 drop of water on specimen
4) Coverslip at 45° angle

1) Place a drop of wet specimen on slide
2) Coverslip at 45° angle

General Tips!
Do NOT mix up pipettes. Use the designated pipette for each solution.

How to clean slides:
wash in sink
throw specimen away
wipe with kimwipe
throw away kimwipe and paper towels in black trash can

Work FAST! Be Efficient!
If there are a shortage of equipment/supplies, go on to a different exercise!
There is only time to get data quickly and jot down notes.
Answer most questions after lab
write these down in your manual!
write these down in your manual!
The basis of BIOL 111 lab
1 & 2
Exercise 3
4, 5, 6, 7
Exercise 8
Tips and Guidelines
- cell wall present (rigid)
- usually rectangular shaped
- one large central vacuole
- chloroplasts present

Plant vs. Animal Cells
- no cell wall (only cell membrane, fluidlike)
- round irregular shaped
- centrioles present
- no chloroplast
- no large central vacuole (smaller ones)
- Bacteria and Archaea
- no nuclear membrane
- usually unicellular
- no membrane-bound organelles
Prokaryotes vs. Eukaryotes
- Plants and Animals
- nucleus
- unicellular or multicellular
- membrane-bound organelles
Exercise VI:
Smaller Elodea leaves at the top of the plant is easier to observe!
Use the water that the Elodea is settled in for your wet mount
Answer Question # 1 before drawing your FOV
Exercise VII:
Get amoeba and paramecium from the very bottom of the jar where they settle.
Do not mix the solution!
Exercise VIII:
How to get onion epidermis
Slice the thinnest potato slice ever imaginable
Only used toothpicks go into biohazard. Everything else goes to trash!
Tips for Exercises!
Field of View
(FOV) sheets
Many labs require FOV drawing

This week's FOVs:
1) Oscillatoria (Ex V)
2) Elodea (Ex VI)
3) Paramecium (Ex VII)
4) Amoeba (Ex VII)
5) Janus Green B [Onion] (Ex VIII)
6) Iodine [Potato] (Ex VIII)
7) Methylene Blue or Neutral Red
[Onion/Cheek] (Ex VIII)
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