Hieu Pham

T/TH 9:00 – 11 :40

Lab 3:
A. Understanding pH and buffers
B. Introduction to the brightfield microscope
Postlab
2.
How does the titration curve of the unbuffered sucrose solution compare in appearance to
the titration curve of both of the buffered solutions?
The titration curve of the unbuffered sucrose solution is linear in comparison to the titration
curves of the other buffered solutions. Without a buffer, the pH of the sucrose becomes
susceptible to change following the addition of an acid or base. Therefore, just one mL of
HCL/NaOH was needed to alter the pH levels above 11.0/below 3.0.
3.
What is the buffering range of the sucrose solution with acetate buffer?
The buffering range for the acetate buffer is approximately 4-6 pH. During this range, the pH of
the solution remains at a steady incline/decline. However, the solution’s pH levels changes
dramatically with each consecutive drop of HCL/NaOH afterwards.
4.
What are the buffering ranges of the sucrose solution with bicarbonate buffer?
The buffering ranges for the sucrose solution with bicarbonate added are approximately 6 - 6.8
and 9 – 10.5 pH. There is a steady slope in between this pH range, before it begins to spike
(indicating that the buffer has reached its limit).
5.
Which solution, if any, seems to have the greater buffering capacity?
The acetate buffer appears to have the greater buffering capacity, seeing as how it’s able to resist
change for a longer amount of added acid/base. With the acetate buffer, it took 8 mL of HCL to
reduce the pH below 3, and 10 mL of NaOH to increase the pH pass 11. Compared to the
bicarbonate buffer, which took 7 mL (1 mL less) of HCL to get the pH below 3, and 3 mL (7 mL
less) of NaOH to boost the pH above 11.
6.
Would either the acetate buffer or the bicarbonate buffer you prepared in this lab be a
good choice for studying biomolecules normally found in human blood?
The buffer ranges for both solutions fall outside of the pH range of blood (7.35 – 7.45)
Therefore, neither of my solutions; bicarbonate, nor acetate would work as an effective buffer for
study as the pH would change too drastically.
Microscope Orientation

1.
You want to better examine the internal structure of a single Euglena cell under a
microscope. Based upon your experience in the lab, which microscope objective would you
use.

To study the organelles of the Euglena cell, I used the 40x objectives (the highest power lens on
the microscope). Enabling a higher magnification of the specimen, I was able to examine and
sketch the internal structures of the Euglena. Furthermore, using the 40x objective grants a
smaller depth of field compared to the others; this allows me to focus on a single cell for study
instead of an array of large groups.
2.
You want to watch several Euglena swimming in the same microscope field of view. Based
upon your experience in the lab, which microscope objective would you use.
When I used the 4x objective lens, I was able to observe a group of Euglena cells all at once. The
4x objective has the largest depth of field out of the other lens, therefore giving me a view of the
bigger picture.
3.
Which objective has the smallest depth of field: the 4X, 10X or 40X objective? Why would
a smaller depth of field be useful when observing cells under a microscope?
The 40x objective has the smallest depth of field amongst the group. A smaller depth of field
enables the user to focus closely on a single unit, and ignore the rest. Therefore, in order to
observe the internal structures of a cell, you would want to minimize your depth to get a better
contrast of the organelles.
4.
You are watching a single Euglena move the right to the left of your field of view. What
direction would you move the slide on the stage to keep the cell in the middle of your field
of view?
Since the Euglena is moving from right to left, you would want to move the slide to the right to
keep the Euglena in your field of view.
5.
Describe two different advantages of viewing Euglena on a prepared slide as opposed to
Euglena on a wet mount slide.
The Euglena are motionless on a prepared slide, so it’s easier to observe them, in contrast to wet
mount slides in which they are still moving around. Moreover, a prepared slide is ready to be
used, whereas you would have to prepare a wet slide (extract specimen, place it on slide, apply
cover, etc.). Sometimes, a wet slide can turn out unfavorable, so you have to spend more time
making a new one.
6.
Describe two different advantages of viewing Euglena on a wet mount slide as opposed to
Euglena on a prepared slide.
An advantage of viewing Euglena cells on a wet mount slide is that you’re able to observe them
going through their natural motions. Furthermore, since the specimens are live, their colors and
qualities are fresh as opposed to the prepared slides where discoloration/deterioration may occur
over time.