Immunocytochemistry procedure: 1. Block _____ binding with protein solution 2. Incubate with _____ antibody 3. Incubate with secondary antibody that contains a ______ (ie collodial gold, horseradish peroxidase)

1. Non-specific protein 2. Primary 3. Visualization label

Lecture 1 Flashcards by Brendan Knapp

Size of RBC

7 um

It acts as internal standard

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resolution of LM

TEM/Scanning

.2 um

1-2 nm

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Hematoxylin is __philic and stains _____

Eosin is ____philic and stains ____

baso, blue/purple

acido, pink/red

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Fixation works by the aldehyrderactive group cross linking____
It mainly preserves ____ but not _____

proteins

proteins, lipids

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Fixation in TEM samples is done with ____

Post fixation is done with ____

glutaraledehyde

osmium tetroxide

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SEM uses the same fixation as TEM, but is dehydrated with ETOH to reach the _____

critical point

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Immunocytochemistry procedure:

Block _____ binding with protein solution
Incubate with _____ antibody
Incubate with secondary antibody that contains a ______ (ie collodial gold, horseradish peroxidase)

Non-specific protein
Primary
Visualization label

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Examples of things stained blue by Hematoxylin (3)

Chromatin, rER, Nucleus, proteins

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Examples of things stained pink with Eosin (2)

Cytoplasm, collagen fibers

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Periodic Acid Schiff Stain-color is ____
Stains mainly ______
Example of staining in cells

magenta
carbohydrates and glycogen
basement membrane of bowman’s capsules; also mucin in trachea

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Silver Stain
colors \_\_\_\_\_ (an example of Type _ collagen)

reticular fibers, type 3

common in lymph nodes

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4 primary tissues

epithelium, connective tissue, muscle, nervous tissue

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Cell Plasma membrane can be seen with a ____ stain.

It has a __laminar binding cheme

osmium trioxide
tri

can only be seen with TEM

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Glycolipids function in ____ interactions and also as _____

cell-cell, receptors

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Gangliosides are glycolipids with _____ acid and are abundant in _____.

sialic acid, nervous tissue.

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GM1 Ganglioside is an example of a cell surface receptor for ___ toxin

Cholera

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Glycolipids are assymetrically distributed in the ____ membrane leaflet

Outer

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Glycocalyx is a ____ coat associated with outer leaflet of plasma membrane. It has a _____ appearance in TEM.

sugar

fuzzy

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The glycocalyx contains cell surface ______, which consist of membrane integral proteins bound to ____.

proteoglyans

glycosaminoglycans

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3 functions of glycocalyx

cellular attachment to extracellular matrix components
binding of antigens/enzymes at cell surface
cell-cell interaction and recognition.

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Cholesterol adds (rigidity or fluidity) to the plasma membrane?

rigidity

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2 basic types of membrane proteins

_____-which can’t be removed without disrupting lipid bilayer
_____-which can be removed w/o disrupting lipid bilayer

Integral

Peripheral

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___ proteins extend into the lipid bilayer

Integral

______ proteins are an example of integral proteins and span the entire membrane. Function as receoptors/transport proteins

transmembrane

Integral proteins are often ____proteins

glyco

We use the ____-fracture method to see membrane proteins

Freeze | Integral proteins remain attatched to P-face

____ proteins do not extend into lipid bilayer and function as part of the ___ or the ____ messenger system.

Peripheral | cytoskelton, intracellular

Membrane fluidity increases with a ___ in temperature and by a greater number of ______ fats in the membrane

rise | unsaturated

Glucose moves across an RBC membrane down a concentration gradient via a ____

glucose transporter protein (GLUT1) it allows D-glucose but not L glucose

The Na/K pumps 3 ___ ions out of the cell and 2 ___ into the cell, against a concentration gradient

Na, K A single ATP is hyrdolyzed to complete this step.

The NaK pump acts to maintain cell volume by ____ the ion concentration intracellularly.

reducing

Inhibitor of ion transport and Na/K ATPase

Oubain

____ can be coupled to the movement of sodium down a concentration gradient

Glucose ie, across intestinal epithelium

Overexpression of ATPase can lead to _____ to cancer drugs

resistance

Cystic Fibrosis Transmembrane Regulator is a ATP and CAMP sensitive ____ ion channel.

Chloride

Membrane receptors are primarily _____. They regulate the ____ of ion channel proteins.

glycoproteins | conformation

Nicotinic Acetylcholine receptors are an example of _____ linked receptors. ___ Venom is an example of a substance that inhibits this channel

ion channel | Snake

Interaction of a signalling molecoule and a __ protein results in activation of intracellular secondary messengers, the most common of which are ___ and ___

G Ca2+, CAMP

Heat Shock Proteins are associated with ____ receptors in the cytosol

Glucocorticoid

____ channel blockers are used for hypertension and cardiovascular disease. ___ channel blockers are used for epilepsy. ____ channel blockers are used for diabetes

Ca Na K

Tetrodotoxin inactivates ___ channels, causing dizziness, respiratory failure, and death.

IgG antibodies binding to TSH receptors stimulates thyroid follicular cells. This is the cause of _____

Graves Disease

_____ is a heriditary condition due to abnormal carrier proteins that cannot remove cystine from the urine. It results in kidney stones

cystinurea

In specimen preparation for TEM, the sections are stained with _____ and ____ salts to provide contrast in the microscope

uranium, lead

_____ electrons are low energy electrons emitted from the surface of a specimen in SEM and these contain surface information.

Secondary

In metachromatic staining, _____ will show up as blue/purple.

polyanions

The diptheria toxin receptor is a _____

glycolipid

The _____ is made up of polar oligosaccharaide side chains covalently linked to protein and glycolipid constituents of the plasma membrane.

Glycocalyx

In Freeze Fracture EM, the _-face is the internal surface of the outer leaflet, while the _-face is the external surface of the inner leaflet.

E | P

_____ molecules and _____ uncharged polar molecules can pass through plasma membrane by simple diffusion.

Hydrophobic, small

Na and Cl- are 10-20x _____ outside the cell than insidfe the cell, while K is 20-40x _____ inside the cell than outside.

higher | higher

Cystic fibrosis is due to defective _____ in epithelial cell of lung and other tissues.

CFTR

Ion channels influence the rate of ____ but not the _____.

flow | direction

Membrane receptors are primarily ____ located on cell surface that bind specific signalling molecoules.

glycoproteins

Cortisol diffuses through the cell membrane, binds to ______ receptors in the cytoplasm, and releases Heat Shock Proteins.

glucocorticoid