Cell Staining and Cytoskeleton

Flashcards based on lecture notes about cell staining techniques, types of stains, cell preparation, cytoskeleton components, and experimental protocols for cell visualization.

Why is cell staining used?

To enhance visualization of the cell or certain cellular components under a microscope by increasing contrast.

What are the purposes of cell staining?

Enhances visualization, highlights metabolic processes, differentiates between live and dead cells, and enumerates cells.

What characteristics of cell samples can be examined through microscopic examination of stained cells?

Size, shape, arrangement of organelles, and external appendages.

What is the difference between in vivo and in vitro cell staining?

In vivo staining is performed on living biological matter in its biological context, while in vitro staining is performed on non-living, fixed biological matter that has been removed from its biological context.

What are the three most popular detection methods in cell staining?

Chromogenic dyes, fluorescent molecules, and antibody staining.

Name some examples of chromogenic dyes used in cell staining.

Bismarck Brown, Carmine, Coomassie blue, Eosin, Fuchsine, Hematoxylin, Iodine, Malachite green, Methylene blue, Neutral/Toluylene red, and Osmium tetroxide.

Name some examples of fluorescent dyes used in cell staining.

DAPI, LysoTracker Green®, MitoTracker Green®, Wheat Germ Agglutinin (WGA), and Phalloidin.

What is the difference between primary and secondary antibodies in antibody staining?

Primary antibodies bind directly to the antigen, while secondary antibodies target another (primary) antibody that is bound to an antigen.

What are some common fixatives used in cell preparation?

Formaldehyde, ethanol, methanol, and picric acid.

What are the general steps of preparing cells for in-vitro staining?

Fixation, permeabilization, staining, and mounting.

What are the three types of filaments that compose the cytoskeleton?

Actin filaments, intermediate filaments, and microtubules.

What are the two most common members of the tubulin family?

Alpha-tubulin and beta-tubulin.

What is the function of phalloidin in relation to actin filaments?

It selectively binds and stabilizes polymerized, filamentous actin without binding monomeric actin.

What is the overall goal of the experimental protocol described in the notes?

To visualize proteins in cells via immunofluorescence, stain actin filaments, and stain the nuclei of cells.

What are the two different cell types used in the experimental protocol?

Dictyostelium cells and L-fibroblasts.

What are some of the key materials used in the experimental protocol?

Paraformaldehyde, Triton X-100, TBST Buffer, Phosphate Buffer Saline (PBS), Anti-alpha tubulin, Goat anti-mouse conjugated to Rhodamine, Fluorescein-Phalloidin conjugate, Mounting media with DAPI.