BIO 208L Lecture Notes - Lecture 2: Immunofluorescence, Autofluorescence, Rhodamine
BIO 206L: Exercise 3 Lab Analysis
Please answer the following questions during lab. Keep in mind that you need to work independently. Write in
complete sentences, using proper grammar and spelling when an explanation is required. Be complete yet
concise in your answers. Submit your report as a pdf file on Canvas before the beginning of the next lab.
1) Create a numbered list of the specimens (e.g., cheek cell, diatoms, etc.) you examined during Exercise 3.
For each specimen, describe the type(s) of microscopy you used to examine the specimen. Include images of
each specimen- indicate magnification and the type of microscopy used.
2) Use the table below to describe the advantages and disadvantages of the following techniques.
Technique
Advantages
Disadvantages
Brightfield with Stain
Phase Contrast
Darkfield
Fluorescence
3) During Procedure 3.2 (preparation of a wet mount of human cheek cells), you added FIX and PERM
solutions to your sample of cheek cells. What does each solution contain? What is the purpose of each
solution? What might happen if you forgot to add the PERM solution?
4) You observed your cheek cells using fluorescent stains. Which two stains were used to prepare these cells?
Which wavelengths of light were used to excite each stain? Which wavelength of light were emitted by each
stain? What is the relationship between excitation and emission wavelengths?
5) What cellular components were visible in your cheek cells? Include a composite image exhibiting both
stains simultaneously. Label the stained cellular structures. Why were other areas in the cell black?
6) You observed BPAE cells using fluorescent microscopy. What stains were used to prepare these cells?
Which wavelengths of light were used to excite each stain? Which wavelengths of light were emitted by each
stain? What is the purpose of the fluorescent filter cube turret?
7) What cellular components were visible in the BPAE cells? Which fluorophore was used to identify the
location of each structure? Include a composite image exhibiting all three stains simultaneously. Label the
cellular components that were stained.
8) Give an example from lab and then compare autofluorescence, direct fluorescence, and
immunofluorescence. Explain how rhodamine phalloidin is different.
9) HeLa cells are very important for medical research. Give an example of how these cells are used by
researchers. Then find an image of HeLa cells that have been fluorescently labeled. Include the image in your
assignment. What types of fluorophores were used to stain the cells? What cellular structures were stained?
Be sure to cite your sources.
find more resources at oneclass.com
find more resources at oneclass.com
Document Summary
Keep in mind that you need to work independently. Write in complete sentences, using proper grammar and spelling when an explanation is required. Submit your report as a pdf file on canvas before the beginning of the next lab: create a numbered list of the specimens (e. g. , cheek cell, diatoms, etc. ) you examined during exercise 3. For each specimen, describe the type(s) of microscopy you used to examine the specimen. Include images of each specimen- indicate magnification and the type of microscopy used: use the table below to describe the advantages and disadvantages of the following techniques. Fluorescence: during procedure 3. 2 (preparation of a wet mount of human cheek cells), you added fix and perm solutions to your sample of cheek cells. What might happen if you forgot to add the perm solution: you observed your cheek cells using fluorescent stains. Include a composite image exhibiting both stains simultaneously.
