Cell Bio Lab Midterm

What did hemacytometer count originally

Blood cells

What do you wipe oil on a microscope lens with

Lens paper

If you count 20 cells in a 1:100 dilution, what would the equation be

(20/4) x 10^4 × 10

Mammalian cell culture advantages

• like in vivo

• scale, can do numerous cells

• control environment

• characterization and homogeneity of sample

Mammalian cell culture disadvantages

• quantity and cost

• not quite like in vivo

• sterile technique

How do CHO cells grow in flask

They grow on the substrate using focal contacts which require calcium

Function of EDTA

Calcium chelator that binds to calcium inactivating cell adherant proteins

Function of trypsin

Protease that digests cell asherant proteins

Function of PBS

Washes the cells to get rid of unwanted serum, media, proteins, etc.

Function of F12K

Medium for CHO, to give them nutrients and environment

What is subcellular localization of proteins

Find where proteins are or the function

Two methods of subcellular localization

1. Fractionation

2. Fluorescence microscopy

What is subcellular fragmentation

Cells get lysed into parts, then centrifuged into pure organelles. Then you can western blot it to find protein of interest

What does it mean when a compound is fluorescent

Absorbs at one wavelength, emits at a longer one

Function of Mitobrilliant 646

Dyes mitochondria red

Function of phalloidin

Dyes actin green

Function of DAPI

Dyes nuclei blue

What do you need to do before using reagents

First fix the cell using PFA, then permeate it using Triton X-100, which takes the proteins and lipids from the cell membrane

Cytoskeleton parts

1. Microfilaments

2. Intermediate filaments

3. Microtubules

Microfilaments

component of contractile ring that pinches cytoplasm apart making two daughter cells

Intermediate filaments

• maintain cell structure and shape

• form desmosomes

Microtubules

• motor protein trackway

• forms mitotic spindle

• component of cilia and flagella

What is ALS/lou gehrig’s

gradual paralysis in a person

What is Hutchinson-Gilford progeria syndrome

premature aging

Colcemid

impedes formation and growth of microtubules

Cytochalasin A

inhibits actin polymeriazation