Laboratory Study Guide,

Cell Ultrastructure

Type your email address in the space provided.

Type your full name in the space provided.

Read:

Cell Ultrastructure, pp 1-7.

Terms to Know in This Chapter:

nucleus (sing.) [NEW klee us]

nuclei (pl.) [NEW klee eye]

mitochondrion (sing.) [my toe CON dree un]

mitochondria (pl.) [my toe CON dree ah]

plastid (sing.) [PLASS tid]

chloroplast (sing.) [CHLOR oh plast]

centrifuge (sing.) [SIN tree fuge]

pellet (sing.) [PEL it]

G force

cellular respiration [CELL you lar] [ress peh RAY shun]

homogenate [ho MOGE eh nate]

filtrate [FIL trate]

residue [REZ eh due]

supernatant [SUP er NAY tant]

sediment [SED eh ment]

sucrose buffer [SUE krose]

plasmolysis [plaz MOL eh siss]

Waring blender [WARE ing]

cheesecloth

iodine [EYE oh dine]

starch grain

tetrazolium [tet rah ZOLE ee um]

2,3,5, triphenyl-2H-tetrazolium chloride [try FEN ul]

hydrogen acceptor

triphenyl formazan [try FEN ul] [FORE mah zan]

glucose [GLUE kose]

carbon dioxide [die OX ide]

acetoorcein [ah seat oh OR seen]

chromosome (sing.) [CROW moe zome]

glycolysis [gly COL oh siss]

Kreb's cycle

Electron Transport System

pyruvic acid [pie RUE vik]

phosphorylation [fos fore ee LAY shun]

outer membrane

inner membrane

crista (sing.) [CRIS tah]

cristae (pl.) [CRIS tee]

People You Need to Know

Schleiden [SHLY den]

Schwann [SWAN]

Van Leewenhoek [LEE un hook]

What You Need to Know:

You should be able to:

1. outline the process of homogenation, filtration, and centrifugation of cell organelles for study in the lab,
2. explain the layering observed in the centrifuge tube,
3. explain the diagnostic tests for mitochondria, starch grains, and nuclei,
4. explain the use of controls in your experiments,
5. identify starch grains, mitochondria, nuclear fragments, and chloroplasts in the microscope,
6. explain the function of sucrose buffer,
7. explain how tetrazolium is a test for the presence of mitochondria, and
8. give an overview of glycolysis, Kreb's cycle, and Electron Transport System.

Exercises: Fill in the Blank.

1. The material at the bottom in a centrifuge tube that does not empty out is called the .
2. After the pea seeds have been ground by the Waring blender, the material is called the .
3. After filtering the homogenate through the cheesecloth, the material that leaks through the cloth is called the while the material retained in the cheesecloth is called the .
4. There are four main layers after the second centrifuge. The liquid layer at the top of the tube is called the . The next layer is a green layer composed of . The next layer is the brownish layer composed mostly of , and the last layer is a white layer composed mostly of . Which of these layers are composed of the heaviest organelles? Which of these layers is composed of the lightest organelles?

   CATEGORY	TUBE A	TUBE B	TUBE C
   1% tetrazolium	5 ml	5 ml	5 ml
   sucrose buffer	5 ml	0 ml	0 ml
   sediment & buffer	0 ml	0 ml	5 ml
   supernatant	0 ml	5 ml	0 ml
   total volume	10 ml	10 ml	10 ml

   Using the chart above, answer questions 5 and 6.

5. Which tube is the control?
6. Which tube would you think would test positive for mitochondria? .
7. What dye is used to stain nuclei or nuclear material?
8. Which dye is used to stain starch grains? .
9. Glycolysis results in the production of two 3-carbon compounds called .
10. The attachment of a phosphate group to ADP to form ATP is called .

Exercises: Multiple Choice. Select the Best Answer.

1. Which of the following serves as a hydrogen acceptor and is an indicator of mitochondrion activity?
   

   (1) iodine
   

   (2) acetoorcein
   

   (3) tetrazolium
   

   (4) methylene blue
   

   (5) Congo red

2. Where do you find mitocondria in the centrifuged tube?

   (1) supernatant
   

   (2) green layer
   

   (3) brown layer
   

   (4) white layer
   

   (5) residue

3. How did you know the white layer contained starch grains?
   

   (1) They stained red with acetoorcein.
   

   (2) They stained blue, purple or black with iodine.
   

   (3) They stained pink with tetrazolium.
   

   (4) They stained blue, purple or black with acetoorcein.
   

   (5) They stained pink with aceotorcein.

4. What do you call the liquid component at the top of the centrifuge tube?
   

   (1) filtrate
   

   (2) residue
   

   (3) supernatant
   

   (4) pellet
   

   (5) more than one answer is correct.

5. Which particles are the heaviest of organelles?
   

   (1) mitochondria
   

   (2) chloroplasts
   

   (3) nuclear material
   

   (4) starch grains
   

   (5) ribosomes